From rjbuesa <@t> yahoo.com Fri May 1 07:54:10 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri May 1 07:54:14 2009 Subject: [Histonet] Processing Fat for Paraffin In-Reply-To: <490013.63581.qm@web46103.mail.sp1.yahoo.com> Message-ID: <520462.17423.qm@web65704.mail.ac4.yahoo.com> Paula: For me the times (2 h/each), your dependable instrument?and the schedule seem to be OK, but I am not so sure about the Citrisolve because this product (1.4 more expensive than xylene) is made of 91% of water and surfactants + 9% d-limonene and I am not very confident that such a "clearing" agent will be able to allow a proper infiltration on your specimens. At least for these specimens I would use xylene or a mixture of 2-propanol and mineral oil (attached to another e-mail to you). Ren? J. --- On Thu, 4/30/09, Va Paula Sicurello wrote: From: Va Paula Sicurello Subject: [Histonet] Processing Fat for Paraffin To: "HistoNet" , "MSA BB" Date: Thursday, April 30, 2009, 6:22 PM Hello out there in fat processing land, I have been given human fat samples and need to embed them in paraffin. In the past I've used a VIP processor for this and now I have an Autotechnicon (vintage dual model) with a timing wheel. I know I need to process these fatties slowly, my question is--can I use 2 hours per step and have it turn out OK? I have a timing wheel punched out for 2 hour steps. My steps would be alcohols: 70, 80, 95 x 2, 100 x 3, Citrisolv x3, paraffin x2 and another paraffin step under vacuum. Let me know your wise and experienced opinions or protocols. I don't have anything else to use except the 43 year old Autotechnicon so don't even suggest it. You make me feel bad that I can't get my research foundation to buy me something new or even newer. ;-) Stewing in somebody else's fat (eew!), Paula :-) Paula Sicurello VA Medical Center San Diego Veterans Medical Research Foundation (VMRF) Core Research Imaging Center 3350 La Jolla Village Dr., MC151 San Diego, CA 92161 858-552-8585 x2397 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dolores_Fischer <@t> baxter.com Fri May 1 08:05:40 2009 From: Dolores_Fischer <@t> baxter.com (Dolores_Fischer@baxter.com) Date: Fri May 1 08:05:57 2009 Subject: [Histonet] Reuse of citrate buffer Message-ID: Just went to the TriState Meeting in Madison Wisconsin and attended a very good lecture on immunohistochemistry focusing on antigen retrieval and it was stressed that one should not reuse any antigen retrieval solution. It should be made fresh when needed. Dolores The information transmitted is intended only for the person(s)or entity to which it is addressed and may contain confidential and/or legally privileged material. Delivery of this message to any person other than the intended recipient(s) is not intended in any way to waive privilege or confidentiality. Any review, retransmission, dissemination or other use of , or taking of any action in reliance upon, this information by entities other than the intended recipient is prohibited. If you receive this in error, please contact the sender and delete the material from any computer. For Translation: http://www.baxter.com/email_disclaimer From k.fd <@t> live.com Fri May 1 09:17:41 2009 From: k.fd <@t> live.com (Karen Doty) Date: Fri May 1 09:17:46 2009 Subject: [Histonet] OCT and RNA Message-ID: Hi, Some of our researchers want to collect frozen tissue for possible future use for frozen sections or for RNA analysis. We would like to know how freezing in OCT will effect subsequent RNA determinations if it is decided that they need the tissue more for RNA analysis than for frozen sections for morphology or IHC studies. Thanks very much for you adivice. Karen _________________________________________________________________ Hotmail? has a new way to see what's up with your friends. http://windowslive.com/Tutorial/Hotmail/WhatsNew?ocid=TXT_TAGLM_WL_HM_Tutorial_WhatsNew1_052009 From JimR0712 <@t> comcast.net Fri May 1 09:18:36 2009 From: JimR0712 <@t> comcast.net (JimR0712@comcast.net) Date: Fri May 1 09:18:41 2009 Subject: [Histonet] Credentials for performing IHC staining In-Reply-To: <122583931.4301601241187381827.JavaMail.root@sz0098a.emeryville.ca.mail.comcast.net> Message-ID: <1053372220.4302421241187516938.JavaMail.root@sz0098a.emeryville.ca.mail.comcast.net> Wew are having an ongoing debate as to the required credentials for a person to perform IHC staining on the Ventana instruments. Can you help ?? Thanks. From rjbuesa <@t> yahoo.com Fri May 1 09:24:50 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri May 1 09:24:54 2009 Subject: [Histonet] OCT and RNA In-Reply-To: Message-ID: <524127.59991.qm@web65716.mail.ac4.yahoo.com> Karen: Usually you will freeze your samples at -20?C and later you will store at -80?C. The answer to your question will have to be provided by your researchers:? is RNA preserved at -80?C for long periods of time? They should make a bibliography research to answer that question. For sure it will preserve the tissue?for morphology and IHC. Ren? J. --- On Fri, 5/1/09, Karen Doty wrote: From: Karen Doty Subject: [Histonet] OCT and RNA To: histonet@lists.utsouthwestern.edu Date: Friday, May 1, 2009, 10:17 AM Hi, Some of our researchers want to collect frozen tissue for possible future use for frozen sections or for RNA analysis. We would like to know how freezing in OCT will effect subsequent RNA determinations if it is decided that they need the tissue more for RNA analysis than for frozen sections for morphology or IHC studies. Thanks very much for you adivice. Karen _________________________________________________________________ Hotmail? has a new way to see what's up with your friends. http://windowslive.com/Tutorial/Hotmail/WhatsNew?ocid=TXT_TAGLM_WL_HM_Tutorial_WhatsNew1_052009_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Fri May 1 09:27:36 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri May 1 09:27:41 2009 Subject: [Histonet] Credentials for performing IHC staining In-Reply-To: <1053372220.4302421241187516938.JavaMail.root@sz0098a.emeryville.ca.mail.comcast.net> Message-ID: <589887.66386.qm@web65713.mail.ac4.yahoo.com> Regardless of the instrument, or if IHC is done manually, it is considered a high complexity test and the person doing it should be an accredited or licensed, specially trained, histotechnologist or med tech specialized in IHC which is a new ASCP accreditation level. Ren? J. --- On Fri, 5/1/09, JimR0712@comcast.net wrote: From: JimR0712@comcast.net Subject: [Histonet] Credentials for performing IHC staining To: "Histonet" , "Histonet2" Date: Friday, May 1, 2009, 10:18 AM Wew are having an ongoing debate as to the required credentials for a person to perform IHC staining on the Ventana instruments. Can you help ?? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jennifer.Bull <@t> northwestpathology.com Fri May 1 12:30:33 2009 From: Jennifer.Bull <@t> northwestpathology.com (Bull, Jennifer L.) Date: Fri May 1 12:30:38 2009 Subject: [Histonet] Frozen Tonsil Controls Message-ID: <85760CECEC18444BB95F26D5E88DAEAA22422789D5@hinet2.hinet.org> Does anyone know where to purchase frozen tonsil control tissue for IF's? Thanks! Jennifer Bull Histology Supervisor jennifer.bull@nwpathology.com From kgrobert <@t> rci.rutgers.edu Fri May 1 12:48:01 2009 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri May 1 12:34:45 2009 Subject: [Histonet] OCT and RNA In-Reply-To: References: Message-ID: <49FB35D1.10709@rci.rutgers.edu> I don't think the OCT will affect the RNA in the tissue, but Ambion has a product and resources that will help you confirm that and protect your RNA. Here's the link to the product: http://www.ambion.com/catalog/CatNum.php?AM7030 And on their home page they have a link to their tech support dept. which has articles from the basics of RNA on up. Hope this helps! Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers, the State University of NJ 41 B Gordon Rd Piscataway, NJ 08854 (732) 445-6914 Karen Doty wrote: >Hi, > > Some of our researchers want to collect frozen tissue for possible future use for frozen sections or for RNA analysis. We would like to know how freezing in OCT will effect subsequent RNA determinations if it is decided that they need the tissue more for RNA analysis than for frozen sections for morphology or IHC studies. > > Thanks very much for you adivice. > >Karen > >_________________________________________________________________ >Hotmail? has a new way to see what's up with your friends. >http://windowslive.com/Tutorial/Hotmail/WhatsNew?ocid=TXT_TAGLM_WL_HM_Tutorial_WhatsNew1_052009_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From JMacDonald <@t> mtsac.edu Fri May 1 12:49:32 2009 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri May 1 12:50:05 2009 Subject: [Histonet] Credentials for performing IHC staining In-Reply-To: <589887.66386.qm@web65713.mail.ac4.yahoo.com> Message-ID: Ren?, Can you please share with us the regulation that states this? Thank you, Jennifer MacDonald Rene J Buesa Sent by: histonet-bounces@lists.utsouthwestern.edu 05/01/2009 07:28 AM Please respond to rjbuesa@yahoo.com To Histonet , Histonet2 , JimR0712@comcast.net cc Subject Re: [Histonet] Credentials for performing IHC staining Regardless of the instrument, or if IHC is done manually, it is considered a high complexity test and the person doing it should be an accredited or licensed, specially trained, histotechnologist or med tech specialized in IHC which is a new ASCP accreditation level. Ren? J. --- On Fri, 5/1/09, JimR0712@comcast.net wrote: From: JimR0712@comcast.net Subject: [Histonet] Credentials for performing IHC staining To: "Histonet" , "Histonet2" Date: Friday, May 1, 2009, 10:18 AM Wew are having an ongoing debate as to the required credentials for a person to perform IHC staining on the Ventana instruments. Can you help ? Thanks. _______________________________________________ Histonet mailing list Histonet@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 kaakslis <@t> gmail.com Fri May 1 13:21:22 2009 From: kaakslis <@t> gmail.com (Vlazhs Sokols) Date: Fri May 1 13:21:29 2009 Subject: [Histonet] FISH together with IF Message-ID: <9a093b740905011121w4bee177dl3facbedff6fd424@mail.gmail.com> Dear Histonetters, In the context of Graft-versus-Host-Disease I?m trying to couple immunfluorescence with FISH. I agree, it is a bit daring to do FISH and IF at once. The problem is that IF is suffering from the pre-treatment during FISH. After 2 failed stainings I was able to state that primary antibodies of IF couldn?t bind antigens after FISH. The question that rose concerning this over-digestion of antigens is ? could I omit the pre-treatment with pepsin, but still but still get a satisfying FISH? Could you, please, look through my protocol. Maybe I?m making the same mistakes you have also encountered. With kind regards, Wladislav Sokalskiy, medical student at the University of Mainz, Germany DAY 1: 1. Putting the section in incubator for 10 min at 70 C. 2. Washing in xylol for 4 x 5 min and rehydrogenating for 5 min in Ethanol. 3. 5 min in ddH2O. 4. For antigen retrieval: washing for 20 min at ~ 100 C in Na-Citrat-Buffer (ph 6). 5. 6 min in Washing-Buffer (don?t know exactly what it consists of, because it was delivered within the FISH-kit) 6. Digestion for 2,5 min with Pepsin. 7. 6 min in Washing-Buffer 8. Dehydrogenization with Ethanol 9. Drying for 12 min at 37 C (is it may be better to dry at the room temperature) 10. Hybridizing for 10 min at 75 C and keeping the section overnight at 37 C (I haven?t coverslipped the section as I kept it overnight. Should I? Was it a mistake on my part?) DAY 2: 1. Washing the section for 5 min in TBS (with 0,05 % Tween 20) 2. Blocking the unspecific antigens with TNT (with 0,05% Tween 20) for 30 min 3. Adding primary antibody (diluted 1:100) 4. Keeping incubating overnight in the moisture chamber DAY 3: 1. Washing 2 x 15 min in TBS (with Tween) 2. Adding secondary antibody (Alexa Fluor 488 ? diluted 1:300) and keeping it incubating for 2 h 3. Washing it 2 x 15 min in TBS (with Tween) 4. Adding DAPI & covering the section with glass. From HornHV <@t> archildrens.org Fri May 1 13:57:17 2009 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri May 1 13:57:24 2009 Subject: [Histonet] Credentials for performing IHC staining In-Reply-To: References: <589887.66386.qm@web65713.mail.ac4.yahoo.com> Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6317D83186@EMAIL.archildrens.org> Although IHC is a high complexity test, it is the reporting of results that constitutes the ruling which only applies to pathologists. Not the techs performing the staining. Just as with all histology testing. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 1 Children's Way Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3155 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jennifer MacDonald Sent: Friday, May 01, 2009 12:50 PM To: rjbuesa@yahoo.com Cc: Histonet2; Histonet; KGroeger@USLABS.net; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Credentials for performing IHC staining Ren?, Can you please share with us the regulation that states this? Thank you, Jennifer MacDonald Rene J Buesa Sent by: histonet-bounces@lists.utsouthwestern.edu 05/01/2009 07:28 AM Please respond to rjbuesa@yahoo.com To Histonet , Histonet2 , JimR0712@comcast.net cc Subject Re: [Histonet] Credentials for performing IHC staining Regardless of the instrument, or if IHC is done manually, it is considered a high complexity test and the person doing it should be an accredited or licensed, specially trained, histotechnologist or med tech specialized in IHC which is a new ASCP accreditation level. Ren? J. --- On Fri, 5/1/09, JimR0712@comcast.net wrote: From: JimR0712@comcast.net Subject: [Histonet] Credentials for performing IHC staining To: "Histonet" , "Histonet2" Date: Friday, May 1, 2009, 10:18 AM Wew are having an ongoing debate as to the required credentials for a person to perform IHC staining on the Ventana instruments. Can you help ? Thanks. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ****************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************** The information 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 HornHV <@t> archildrens.org Fri May 1 14:20:31 2009 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri May 1 14:20:35 2009 Subject: [Histonet] Frozen Tonsil Controls In-Reply-To: <85760CECEC18444BB95F26D5E88DAEAA22422789D5@hinet2.hinet.org> References: <85760CECEC18444BB95F26D5E88DAEAA22422789D5@hinet2.hinet.org> Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6317D83187@EMAIL.archildrens.org> I don't know where to purchase them but if you have a childrens hospital nearby they can probably accommodate you. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 1 Children's Way Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3155 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bull, Jennifer L. Sent: Friday, May 01, 2009 12:31 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Frozen Tonsil Controls Does anyone know where to purchase frozen tonsil control tissue for IF's? Thanks! Jennifer Bull Histology Supervisor jennifer.bull@nwpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ****************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************** The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. From mtighe <@t> trudeauinstitute.org Fri May 1 15:04:27 2009 From: mtighe <@t> trudeauinstitute.org (Mike Tighe) Date: Fri May 1 15:04:51 2009 Subject: [Histonet] OCT and RNA Message-ID: <49FB1D90.26E4.00EE.0@trudeauinstitute.org> We have taken sections from OCT embedded tissues and processed them for RNA analysis with out any problems. You should be careful to move the tissue to a solution that will protect the RNA once it has been either sectioned or thawed. Good Luck! Trudeau Institute email service will be temporarily suspended on May 2-3 to complete the joining of electrical services between the newly constructed Stafford Wing and our existing facilities. Email sent to and from all Trudeau Institute employees will be held in queue until services are restored on Monday, May 4th From mitchellja <@t> email.chop.edu Fri May 1 15:13:46 2009 From: mitchellja <@t> email.chop.edu (Janice Mitchell) Date: Fri May 1 15:14:19 2009 Subject: [Histonet] STP420 Message-ID: <49FB1FBA020000000051F658@email.chop.edu> Good Afternoon, We have the thermofisher stp 420 processor. We are having major cross contamination problems. Finding brain and placenta fragments in with other tissue. Anyone else with this processor having similar problems? Thanks, Janice From mpence <@t> grhs.net Fri May 1 15:41:37 2009 From: mpence <@t> grhs.net (Mike Pence) Date: Fri May 1 15:41:42 2009 Subject: [Histonet] STP420 In-Reply-To: <49FB1FBA020000000051F658@email.chop.edu> Message-ID: <661949901A768E4F9CC16D8AF8F2838C017A3B2E@IS-E2K3.grhs.net> I have never used this tissue processor, but I don't understand how it would be the tissue processor that would be causing the contamination. Does this processor have different processing chamber? If so, does it use the same processing solutions? Is all your tissues sectioned on the same cutting board? By the same cutter? Just throwing out some general thoughts. Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Janice Mitchell Sent: Friday, May 01, 2009 3:14 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] STP420 Good Afternoon, We have the thermofisher stp 420 processor. We are having major cross contamination problems. Finding brain and placenta fragments in with other tissue. Anyone else with this processor having similar problems? Thanks, Janice _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jmjohnson34 <@t> hotmail.com Fri May 1 16:00:53 2009 From: jmjohnson34 <@t> hotmail.com (Jennifer Johnson) Date: Fri May 1 16:00:56 2009 Subject: [Histonet] (no subject) Message-ID: Can anyone tell me how to use liquid nitrogen on fatty sections and also give me a source for purchasing it and a rough estimate of how much it costs. I would appreciate any help. We have been battling difficult breast specimens at our tiny little (cheap) hospital for entirely too long. Thanks, Jennifer Johnson, HTL,(ASCP) _________________________________________________________________ Rediscover Hotmail?: Get e-mail storage that grows with you. http://windowslive.com/RediscoverHotmail?ocid=TXT_TAGLM_WL_HM_Rediscover_Storage2_042009 From histoman3000 <@t> live.com Fri May 1 16:47:25 2009 From: histoman3000 <@t> live.com (Jimmy Markum) Date: Fri May 1 16:47:30 2009 Subject: [Histonet] Cancer Diagnostics and Surgipath Message-ID: I purchased Surgipath cassettes from Cancer Diagnostics but they do not supply these any more and I cannot reach my rep. Does anyone know why? _________________________________________________________________ Windows Live? SkyDrive?: Get 25 GB of free online storage. http://windowslive.com/online/skydrive?ocid=TXT_TAGLM_WL_skydrive_042009 From pinetree110567 <@t> yahoo.com Fri May 1 16:53:13 2009 From: pinetree110567 <@t> yahoo.com (Jack Cates) Date: Fri May 1 16:53:18 2009 Subject: [Histonet] RE: Cancer Diagnostics and Surgipath Message-ID: <533538.51429.qm@web110816.mail.gq1.yahoo.com> This may be just a rumor, but I heard Patrick, one of the Surgipath Sales Managers, actually set up Cancer Diagnostics and was running a side business while also still working for Surgipath. And when Surgipath was sold to Leica, Leica in doing their due diligence discovered this and summilarily dismissed him and thus cut him off of supply for stealing leads and may be taking legal action. JC. > From: histoman3000@live.com > To: histonet@lists.utsouthwestern.edu > Date: Fri, 1 May 2009 17:47:25 -0400 > Subject: [Histonet] Cancer Diagnostics and Surgipath > > > I purchased Surgipath cassettes from Cancer Diagnostics but they do not supply these any more and I cannot reach my rep. > Does anyone know why? From gu.lang <@t> gmx.at Sat May 2 04:53:02 2009 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat May 2 04:53:12 2009 Subject: AW: [Histonet] FISH together with IF In-Reply-To: <9a093b740905011121w4bee177dl3facbedff6fd424@mail.gmail.com> References: <9a093b740905011121w4bee177dl3facbedff6fd424@mail.gmail.com> Message-ID: <67B9552B5FD34EDDB670704B63467969@dielangs.at> Hi Wladislav, this technique is called FICTION. I'm sure, that you have done a literature search on this. If you need pepsin for your FISH depends on the fixation-status of the tissue for permeabilization and digestion of the histon-proteins. I guess formaldehyde fixed tissue cannot do without it. I have no personal experience with this combined method. Perhaps you have to omit one of the pretreatment steps of FISH. I would try it. Slide drying: I would dry at RT or just 1-2 min at 37?C. For immunohistochemistry you have to avoid dry tissue - hmm. Coverslipping: and sealing with rubbercement (Fixogum)is always recommended for denaturation and hybridization over night. (but if you have got signals, who knows) Perhaps my thoughts were helpful Gudrun -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Vlazhs Sokols Gesendet: Freitag, 01. Mai 2009 20:21 An: Histonet@lists.utsouthwestern.edu Betreff: [Histonet] FISH together with IF Dear Histonetters, In the context of Graft-versus-Host-Disease I?m trying to couple immunfluorescence with FISH. I agree, it is a bit daring to do FISH and IF at once. The problem is that IF is suffering from the pre-treatment during FISH. After 2 failed stainings I was able to state that primary antibodies of IF couldn?t bind antigens after FISH. The question that rose concerning this over-digestion of antigens is ? could I omit the pre-treatment with pepsin, but still but still get a satisfying FISH? Could you, please, look through my protocol. Maybe I?m making the same mistakes you have also encountered. With kind regards, Wladislav Sokalskiy, medical student at the University of Mainz, Germany DAY 1: 1. Putting the section in incubator for 10 min at 70 C. 2. Washing in xylol for 4 x 5 min and rehydrogenating for 5 min in Ethanol. 3. 5 min in ddH2O. 4. For antigen retrieval: washing for 20 min at ~ 100 C in Na-Citrat-Buffer (ph 6). 5. 6 min in Washing-Buffer (don?t know exactly what it consists of, because it was delivered within the FISH-kit) 6. Digestion for 2,5 min with Pepsin. 7. 6 min in Washing-Buffer 8. Dehydrogenization with Ethanol 9. Drying for 12 min at 37 C (is it may be better to dry at the room temperature) 10. Hybridizing for 10 min at 75 C and keeping the section overnight at 37 C (I haven?t coverslipped the section as I kept it overnight. Should I? Was it a mistake on my part?) DAY 2: 1. Washing the section for 5 min in TBS (with 0,05 % Tween 20) 2. Blocking the unspecific antigens with TNT (with 0,05% Tween 20) for 30 min 3. Adding primary antibody (diluted 1:100) 4. Keeping incubating overnight in the moisture chamber DAY 3: 1. Washing 2 x 15 min in TBS (with Tween) 2. Adding secondary antibody (Alexa Fluor 488 ? diluted 1:300) and keeping it incubating for 2 h 3. Washing it 2 x 15 min in TBS (with Tween) 4. Adding DAPI & covering the section with glass. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Sat May 2 05:04:40 2009 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat May 2 05:04:47 2009 Subject: AW: [Histonet] FISH together with IF In-Reply-To: <9a093b740905011121w4bee177dl3facbedff6fd424@mail.gmail.com> References: <9a093b740905011121w4bee177dl3facbedff6fd424@mail.gmail.com> Message-ID: <01EC940A46C241FB803EF41028875770@dielangs.at> I've just found a publication, where the IF is done before FISH. Look at this: http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=1770804&blobtype=pdf bye Gudrun -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Vlazhs Sokols Gesendet: Freitag, 01. Mai 2009 20:21 An: Histonet@lists.utsouthwestern.edu Betreff: [Histonet] FISH together with IF Dear Histonetters, In the context of Graft-versus-Host-Disease I?m trying to couple immunfluorescence with FISH. I agree, it is a bit daring to do FISH and IF at once. The problem is that IF is suffering from the pre-treatment during FISH. After 2 failed stainings I was able to state that primary antibodies of IF couldn?t bind antigens after FISH. The question that rose concerning this over-digestion of antigens is ? could I omit the pre-treatment with pepsin, but still but still get a satisfying FISH? Could you, please, look through my protocol. Maybe I?m making the same mistakes you have also encountered. With kind regards, Wladislav Sokalskiy, medical student at the University of Mainz, Germany DAY 1: 1. Putting the section in incubator for 10 min at 70 C. 2. Washing in xylol for 4 x 5 min and rehydrogenating for 5 min in Ethanol. 3. 5 min in ddH2O. 4. For antigen retrieval: washing for 20 min at ~ 100 C in Na-Citrat-Buffer (ph 6). 5. 6 min in Washing-Buffer (don?t know exactly what it consists of, because it was delivered within the FISH-kit) 6. Digestion for 2,5 min with Pepsin. 7. 6 min in Washing-Buffer 8. Dehydrogenization with Ethanol 9. Drying for 12 min at 37 C (is it may be better to dry at the room temperature) 10. Hybridizing for 10 min at 75 C and keeping the section overnight at 37 C (I haven?t coverslipped the section as I kept it overnight. Should I? Was it a mistake on my part?) DAY 2: 1. Washing the section for 5 min in TBS (with 0,05 % Tween 20) 2. Blocking the unspecific antigens with TNT (with 0,05% Tween 20) for 30 min 3. Adding primary antibody (diluted 1:100) 4. Keeping incubating overnight in the moisture chamber DAY 3: 1. Washing 2 x 15 min in TBS (with Tween) 2. Adding secondary antibody (Alexa Fluor 488 ? diluted 1:300) and keeping it incubating for 2 h 3. Washing it 2 x 15 min in TBS (with Tween) 4. Adding DAPI & covering the section with glass. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMyers1 <@t> aol.com Sat May 2 08:34:18 2009 From: JMyers1 <@t> aol.com (JMyers1@aol.com) Date: Sat May 2 08:36:11 2009 Subject: [Histonet] Reusing Citrate Antigen Retrieval Buffer Message-ID: Jason: In my opinion, any recommendation to re-use a retrieval buffer just isn't 'good science'. The problem is that one cannot know, with any certainty, when a solution loses it effectiveness. The primary reason that the re-use approach has been advocated is because this device consumes a disproportionate volume of reagent for the number slides that it can handle (when 'full'). In addition to this 'scientific' issue, there is the issue of cost. Fundamentally, retrieval buffers are (or should be) 'cheap' -- that is, they cost only a few cents per mL, so, depending on the quantity and arrangement of slides within a given device, cost-per-slide values range from twenty-five cents to nearly two dollars. Using a PT Module, however, the lowest possible cost-per-slide that one can obtain would be in the best-case scenario of placing 24 slides in each 'tank', but the reality is, one rarely uses completely filled slide racks, so the cost-per-slide value is usually even higher. If you're interested, I can e-mail you (or anyone else) a cost-analysis spreadsheet that will likely surprise you. Finally, for what its worth, most of labs that I've ever spoken with who've (initially) implemented this device have gone back to using a pressure cooker because of cost concerns and the hassle of handling the hot, heavy tanks. Joe Myers, M.S, CT(ASCP) ------------------------------ Message: 6 Date: Thu, 30 Apr 2009 09:09:10 -0600 From: "Jason McGough" Subject: [Histonet] Reusing Citrate Antigen Retrieval Buffer We are looking into using our .01M Citrate Antigen Retrieval Buffer in our Dako PT Link for Antigen Retrieval on our IHC stains. The question that we are wondering about is anybody reusing this solution for multiple times? If so, how many times? What is your dilution? Like the High pH Antigen Retrieval solution from Dako, it is recommended to use up to 3 time before replacement. We are having several problems with the High pH antigen retrieval solution from Dako and want to try Citrate, since that is what we have been using for many years with great results. We previously used the Pascal Pressure Cooker from Dako for Antigen Retrieval but now have the PT Link. Thank you in advance for your replies. Jason McGough HT(ASCP) Account Representative - Anatomic Pathology Clinical Laboratory of the Black Hills **************Eat Great & Lose Weight FASTER! Start the South Beach Diet Online - FREE Profile! (http://pr.atwola.com/promoclk/100126575x1221822996x1201398599/aol?redir=http:%2F%2Fad.doubleclick.net%2Fclk%3B213623126%3B35100424% 3Bk) From sheila_adey <@t> hotmail.com Sat May 2 09:27:11 2009 From: sheila_adey <@t> hotmail.com (sheila adey) Date: Sat May 2 09:27:16 2009 Subject: [Histonet] STP420 In-Reply-To: <661949901A768E4F9CC16D8AF8F2838C017A3B2E@IS-E2K3.grhs.net> References: <49FB1FBA020000000051F658@email.chop.edu> <661949901A768E4F9CC16D8AF8F2838C017A3B2E@IS-E2K3.grhs.net> Message-ID: We were having issues with placenta contamination aswell. Our Dr.s now wrap the placenta sections in tissue paper and this seems to have cut down on the problem. Sheila Adey HT MLT Port Huron Hospital Michigan > Date: Fri, 1 May 2009 15:41:37 -0500 > From: mpence@grhs.net > To: mitchellja@email.chop.edu; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] STP420 > CC: > > I have never used this tissue processor, but I don't understand how it > would be the tissue processor that would be causing the contamination. > Does this processor have different processing chamber? If so, does it > use the same processing solutions? Is all your tissues sectioned on the > same cutting board? By the same cutter? Just throwing out some general > thoughts. > > Mike > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Janice > Mitchell > Sent: Friday, May 01, 2009 3:14 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] STP420 > > > Good Afternoon, We have the thermofisher stp 420 processor. We are > having major cross contamination problems. Finding brain and placenta > fragments in with other tissue. Anyone else with this processor having > similar problems? Thanks, Janice > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ One at a time or all at once? Get updates from your friends in one place. http://go.microsoft.com/?linkid=9660827 From rjbuesa <@t> yahoo.com Sat May 2 09:42:47 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat May 2 09:42:51 2009 Subject: [Histonet] xylene In-Reply-To: Message-ID: <188447.84837.qm@web65707.mail.ac4.yahoo.com> I also used recycled xylene before I switched to mineral oil and?I used?a B/R Corp recycler and never had that problem. The temperature sensor of your recycler probably failed?and started to collect mixed xylene. Try to check the sensor. Ren? J. --- On Tue, 4/28/09, Moffatt, Loretta wrote: From: Moffatt, Loretta Subject: [Histonet] xylene To: "'histonet@lists.utsouthwestern.edu'" Date: Tuesday, April 28, 2009, 2:06 PM We have recently had a problem with poor tissue processing (soft to cut/ poor staining). None of the tissue was processed the first time, and ultimately, some fragile tissue did not survive. The alcohols checked out and the processor, itself, does not appear to be an issue. We used recycled xylene in the processor first, followed by fresh xylene. The xylene, which came from a CBG recycler, failed the test for contamination, after the fact. Has anyone seen this kind of problem? We would appreciate any input. Thank you. Loretta Moffatt, MT(ASCP),MHA Laboratory Manager 777 Rural Avenue Williamsport, PA 17701 570-321-2326 (F)570-321-2489 lmoffatt@susquehannahealth.org Patience is never more important than when you are on the verge of losing it. Confidentiality Notice: This message and any attachments originate by electronic mail from Susquehanna Health System and their subsidiaries/affiliates ("SHS"). Both this document and any attachments are intended for the sole use of the addressee indicated above and may contain proprietary, privileged and/or confidential information. If you are not the intended recipient of this message, you are hereby notified that any use or disclosure of this information is strictly prohibited. If you received this message in error, or have reason to believe you are not authorized to receive it, please notify the sender by reply email, with a copy to ITSecurity@susquehannahealth.org < mailto:ITSecurity@susquehannahealth.org>, and then promptly delete the original and reply messages. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From marktarango <@t> gmail.com Sat May 2 13:09:13 2009 From: marktarango <@t> gmail.com (Mark Tarango) Date: Sat May 2 13:09:17 2009 Subject: [Histonet] STP420 In-Reply-To: <661949901A768E4F9CC16D8AF8F2838C017A3B2E@IS-E2K3.grhs.net> References: <49FB1FBA020000000051F658@email.chop.edu> <661949901A768E4F9CC16D8AF8F2838C017A3B2E@IS-E2K3.grhs.net> Message-ID: <5b6eb13e0905021109jf92d519ia79276024cab4667@mail.gmail.com> That processor spins the tissue around and causes much more agitation than with other tissue processors. Tissue gets out of the cassettes easier. It makes for faster processing, but you have to use mesh cassettes or wrap placenta and other small friable tissues. Mark On Fri, May 1, 2009 at 1:41 PM, Mike Pence wrote: > I have never used this tissue processor, but I don't understand how it > would be the tissue processor that would be causing the contamination. > Does this processor have different processing chamber? If so, does it > use the same processing solutions? Is all your tissues sectioned on the > same cutting board? By the same cutter? Just throwing out some general > thoughts. > > Mike > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Janice > Mitchell > Sent: Friday, May 01, 2009 3:14 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] STP420 > > > Good Afternoon, We have the thermofisher stp 420 processor. We are > having major cross contamination problems. Finding brain and placenta > fragments in with other tissue. Anyone else with this processor having > similar problems? Thanks, Janice > > > > _______________________________________________ > Histonet mailing list > Histonet@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 matthewtclose <@t> gmail.com Sat May 2 14:34:13 2009 From: matthewtclose <@t> gmail.com (Matthew Close) Date: Sat May 2 14:34:17 2009 Subject: [Histonet] re: Xylene Message-ID: <6abc767b0905021234g1e63609sc401c19b821f3818@mail.gmail.com> I don't know whether you are decalcifying tissue or not, but I have had similar problems with tissue that was over-decalcified. It was particularly bad when decalcified with commercial "decalcifying agent" which contained formic along with several chelating agents. The end result was tissue that was soft to cut, stained poorly. I had to alter several protocols to rescue the sections during staining and eventually got pretty good results. Email me if you are interested. And good luck. Matt Close Matthew T. Close Lehigh University, Department of Biological Sciences From rcartun <@t> harthosp.org Sat May 2 18:14:41 2009 From: rcartun <@t> harthosp.org (Richard Cartun) Date: Sat May 2 18:20:03 2009 Subject: [Histonet] Entamoeba Trophozoites Message-ID: <49FC9BA1020000770000B788@gwmail4.harthosp.org> This is a late response, but I have IHC staining for E. histolytica on formalin-fixed, parafffin-embedded tissue in my laboratory. Richard Richard W. Cartun, Ph.D. Director, Histology & Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Stephen Peters M.D." 04/06/09 7:26 AM >>> It is obviously too late for this email to be useful in this case but I will offer a suggestion for future reference. When E histolytica in volves the liver it typically is in the form of an abscess. Look very carefulls at the specimen and make a wet prep smear of a small amount of tissue from anything that looks different than the normal brown appearing liver tissue. If you see anything soft or liquifactive yellowish or tan this would be ideal. Crush a speck of this tissue on a slide with a little saline and examine immediately with a lot of contrast. If E. histolitica are present you will see these cool little buggers, not much bigger than a histiocyte with small cookie cutter round nuckei and I tiny central dot crawling around the slide with purposeful motion and some containg RBCs. The encysted forms are a bit larger, rounder and contain 4 nucleoli. If recieved in saline it would be worth spinning down the saline an having a look at that. You can stain the slides as discribed above after looking at the wet mount. Stephen Peters M.D. 201 847 0052 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 Phone and fax 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jason.PALMER <@t> svhm.org.au Sun May 3 23:12:28 2009 From: Jason.PALMER <@t> svhm.org.au (PALMER Jason (SVHM)) Date: Sun May 3 23:12:36 2009 Subject: [Histonet] Optiprobe question References: Message-ID: Hello. I've just come across a reference (from 1998) where tissue was fixed in "Optiprobe" prior to paraffin embedding. I can't find much basic info on this fixative on the net and was wondering if anyone can tell me anything about it, or whether it is a formaldehyde-based product? Thanks, Jason Jason Palmer Histology Laboratory Coordinator Bernard O'Brien Institute of Microsurgery 42 Fitzroy St, Fitzroy Victoria 3065 Australia tel +61 3 9288 4018 fax +61 3 9416 0926 email: jason.palmer@svhm.org.au Disclaimer : The contents of this e-mail including any attachments are intended only for the person or entity to which this e-mail is addressed and may contain confidential, privileged and/or commercially sensitive material. If you are not, or believe you may not be, the intended recipient, please advise the sender immediately by return e-mail, delete this e-mail and destroy any copies. ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From tifei <@t> foxmail.com Mon May 4 04:27:34 2009 From: tifei <@t> foxmail.com (TF) Date: Mon May 4 04:28:23 2009 Subject: [Histonet] Which kind of pre-coated slides is good for brain sections? Message-ID: <200905041727291805290@foxmail.com> Just want to buy some slides for our brain sections. Pre-fixed before cryostat section. normally 10 um -40 um. Thanks very much ! s/#Disclaimer From histonet.nospam <@t> vneubert.com Mon May 4 04:32:02 2009 From: histonet.nospam <@t> vneubert.com (V. Neubert) Date: Mon May 4 04:32:11 2009 Subject: [Histonet] Destain Hematoxylon Message-ID: <49FEB612.4060200@vneubert.com> Hi, I want to destain hematoxylin (Shandon's Gill II). How to do that? Thanks in advance, V. Neubert From mpence <@t> grhs.net Mon May 4 08:17:12 2009 From: mpence <@t> grhs.net (Mike Pence) Date: Mon May 4 08:17:17 2009 Subject: [Histonet] STP420 In-Reply-To: <5b6eb13e0905021109jf92d519ia79276024cab4667@mail.gmail.com> Message-ID: <661949901A768E4F9CC16D8AF8F2838C017A3B2F@IS-E2K3.grhs.net> That makes sense. Know I can see where the processor may be responsible for the carry over. This is a good thing to know if and when looking for new tissue processors. Mike -----Original Message----- From: Mark Tarango [mailto:marktarango@gmail.com] Sent: Saturday, May 02, 2009 1:09 PM To: Mike Pence Cc: Janice Mitchell; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] STP420 That processor spins the tissue around and causes much more agitation than with other tissue processors. Tissue gets out of the cassettes easier. It makes for faster processing, but you have to use mesh cassettes or wrap placenta and other small friable tissues. Mark On Fri, May 1, 2009 at 1:41 PM, Mike Pence wrote: I have never used this tissue processor, but I don't understand how it would be the tissue processor that would be causing the contamination. Does this processor have different processing chamber? If so, does it use the same processing solutions? Is all your tissues sectioned on the same cutting board? By the same cutter? Just throwing out some general thoughts. Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Janice Mitchell Sent: Friday, May 01, 2009 3:14 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] STP420 Good Afternoon, We have the thermofisher stp 420 processor. We are having major cross contamination problems. Finding brain and placenta fragments in with other tissue. Anyone else with this processor having similar problems? Thanks, Janice _______________________________________________ Histonet mailing list Histonet@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 woods1008 <@t> yahoo.cn Mon May 4 08:54:16 2009 From: woods1008 <@t> yahoo.cn (=?utf-8?B?5LyN5rK7?=) Date: Mon May 4 08:54:25 2009 Subject: [Histonet] Journal of histotechnology paper searching Message-ID: <949125.87395.qm@web92413.mail.cnh.yahoo.com> hello, ????? I?was sarching for a paper from Journal of histotechnology. Unfortunately the paper doesn't show up on medline and I can't seem to? find an archive for J Histotech that goes back that far. I wish?somebody could help me?.? ???? The paper is: ?Okia, Zelda, Tihan, Tarik, Kane, Philip. Clear cell carcinoma of the lung: Use of immunohistochemistry to determine primary vs. metastatic origin. Journal of Histotechnology. 1998,21(2) : 159-164 ??? Thank you! ? ?????????????????????????????????????????????????????????????????????????????????????????????????????? Woods ___________________________________________________________ ????????????????? http://card.mail.cn.yahoo.com/ From dmccaig <@t> ckha.on.ca Mon May 4 08:59:13 2009 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Mon May 4 08:59:19 2009 Subject: [Histonet] ER/PR scoring guidelines Message-ID: Can someone provide me with the guidelines (Canadian) for scoring ER/PR and what the preferred antibody is for determining this. I see there are several clones. Diana From rjbuesa <@t> yahoo.com Mon May 4 09:44:26 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon May 4 09:44:29 2009 Subject: [Histonet] Journal of histotechnology paper searching In-Reply-To: <949125.87395.qm@web92413.mail.cnh.yahoo.com> Message-ID: <389053.90161.qm@web65711.mail.ac4.yahoo.com> If you are a member of the NSH you can ask the article from them and they will send you a pdf file Ren? J. --- On Mon, 5/4/09, ?? wrote: From: ?? Subject: [Histonet] Journal of histotechnology paper searching To: Histonet@lists.utsouthwestern.edu Date: Monday, May 4, 2009, 9:54 AM hello, ????? I?was sarching for a paper from Journal of histotechnology. Unfortunately the paper doesn't show up on medline and I can't seem to? find an archive for J Histotech that goes back that far. I wish?somebody could help me?.? ???? The paper is: ?Okia, Zelda, Tihan, Tarik, Kane, Philip. Clear cell carcinoma of the lung: Use of immunohistochemistry to determine primary vs. metastatic origin. Journal of Histotechnology. 1998,21(2) : 159-164 ??? Thank you! ? ?????????????????????????????????????????????????????????????????????????????????????????????????????? Woods ___________________________________________________________ ????????????????? http://card.mail.cn.yahoo.com/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Montina.VanMeter <@t> pbrc.edu Mon May 4 10:30:18 2009 From: Montina.VanMeter <@t> pbrc.edu (Montina Van Meter) Date: Mon May 4 10:30:29 2009 Subject: [Histonet] 2009 Louisiana Society for Histotechnology Annual Symposium/Convention Message-ID: <4FE7FB862E90E448AE32388E759220E50140A4DF@pbrcas31.pbrc.edu> Hello Histonetters, The Louisiana Society for Histotechnology is pleased to announce the 26th Annual Symposium/Convention: "Your Histeaux Surplus Package" June 12 & 13, 2009 at the Bourbon Orleans Hotel 717 Orleans St. New Orleans, LA 70117 www.bourbonorleans.com The LSH block of rooms will be held until May 11, 2009. For those attendees who may want to visit the beautiful sights and sounds of New Orleans, the Bourbon Orleans Hotel will honor the room rates for three days prior and three days after the meeting. Special on-site parking rates for LSH attendees are available as well as public parking around Jackson Square. For reservations call 1-504-523-2222 or 1-866-513-9744 and mention you are with the LSH group. Do to relocation of many of our members we would ask everyone who would like to receive a brochure/membership form in the mail, email or fax, to please contact Tina Van Meter at 225-603-0953, vanmetmj@pbrc.edu or Dixie Benoit at 337-233-1951, dixiehistochick@bellsouth.net. Walk-ins are always welcome, but pre-registrants will receive a complimentary buffet lunch each day! Please make additional copies of your registration form for co-workers in your lab or facility that might be interested in attending. The LSH would also like to extend the invitation to our fellow technologists and pathologists from surrounding states. We encourage everyone to attend in order to build our networking potential, earn those valuable CEU's and enjoy the beautiful city of New Orleans. We have a variety of topics presented by experienced speakers that promises to benefit everyone. The attendees will have access to several scientific vendor exhibits during the entire symposium. I have listed the workshops below and encourage y'all to come on down to the Bourbon Orleans Hotel in the French Quarter! 2009 LSH State Meeting Workshops: WS#1 - Am I Really Ready for this CAP Inspection? WS#2 - Mouse to Horse: Differences in working with human and animal tissue WS#3 - Breast Cancer and the Standardization of HER2/neu Testing WS#4 - Contemporary Trends in Immunohistochemistry WS#5 - Troubleshooting Routine Special Stains WS#6 - Which Code Do I Use? (CPT Coding) WS#7 - Are You REALLY Ready for the Next Catastrophic Event That Will Affect Your Hospital or City? WS#8 - 2 Crazy Cat Ladies Give Their Opinions of Life, Liberty, and Lab Management Hope to see you in June! Tina Van Meter Montina J. Van Meter, HT (ASCP) Lab Manager Autonomic Neuroscience Pennington Biomedical Research Center 6400 Perkins Rd. Baton Rouge, LA 70791 225-763-2564 From Jessica.Vacca <@t> HCAhealthcare.com Mon May 4 10:08:04 2009 From: Jessica.Vacca <@t> HCAhealthcare.com (Vacca Jessica) Date: Mon May 4 10:35:12 2009 Subject: [Histonet] Lab Safety Message-ID: <938D716CD445614ABBB817517557B6F4C24A4709@NADCWPMSGCMS09.hca.corpad.net> Would anyone be willing to share what they do for lab safety as far as teaching, and keeping up with what is requested by CAP? Since I have chemicals, I have been nominated to be in charge of the Chemical Hygiene manual which now consists of all training for the lab and so forth. What are you doing for the fire extinguishers and annual review of the safety manual to your staff? Do you have a check list you can share? Anything will help. Thanks Jessica Vacca Histology Supervisor 119 Oakfield Dr Brandon Fl 33511 (813) 571-5193 (813) 571-5169 FAX ? From CIngles <@t> uwhealth.org Mon May 4 10:46:25 2009 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Mon May 4 10:46:29 2009 Subject: [Histonet] Oil red o mounting media References: Message-ID: We use Crystalmount. You don't even have to coverslip. Just make sure you evenly distribute the mountant on the slide and let it dry. We usually put it on a hot plate for a bit. This leaves a hardened shell around the tissue. The slides can be read like this too. If you really need a coverslip, it can be quick dipped in xylene and coverslipped with your regular mounting media. You can also coverslip directly with crystal mount, but we find it dries around the section and creates a big air bubble encompassing the tissue. This technique also has the added benefit of keeping the fat (and the stain) in the correct areas and is not squeezed out from the pressure during direct mounting. Claire Hi, I will be staining many samples for Oil red O in the near future. I was wondering if there were any aqueous mounting medias out there that perform superior to Glycergel? Thanks Dave From laurie.colbert <@t> huntingtonhospital.com Mon May 4 10:52:30 2009 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Mon May 4 10:52:38 2009 Subject: [Histonet] xylene Message-ID: <57BE698966D5C54EAE8612E8941D76830580B3D4@EXCHANGE3.huntingtonhospital.com> We had a paraffin plug in the waste container, which caused a malfunction of the unit and contamination of our reagents. We paid for CBG to come out and perform and thorough PM before we used it again. We have had the unit for about 7 or 8 years and this is the first time this happened. Laurie Colbert -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Moffatt, Loretta Sent: Tuesday, April 28, 2009 11:06 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] xylene We have recently had a problem with poor tissue processing (soft to cut/ poor staining). None of the tissue was processed the first time, and ultimately, some fragile tissue did not survive. The alcohols checked out and the processor, itself, does not appear to be an issue. We used recycled xylene in the processor first, followed by fresh xylene. The xylene, which came from a CBG recycler, failed the test for contamination, after the fact. Has anyone seen this kind of problem? We would appreciate any input. Thank you. Loretta Moffatt, MT(ASCP),MHA Laboratory Manager 777 Rural Avenue Williamsport, PA 17701 570-321-2326 (F)570-321-2489 lmoffatt@susquehannahealth.org Patience is never more important than when you are on the verge of losing it. Confidentiality Notice: This message and any attachments originate by electronic mail from Susquehanna Health System and their subsidiaries/affiliates ("SHS"). Both this document and any attachments are intended for the sole use of the addressee indicated above and may contain proprietary, privileged and/or confidential information. If you are not the intended recipient of this message, you are hereby notified that any use or disclosure of this information is strictly prohibited. If you received this message in error, or have reason to believe you are not authorized to receive it, please notify the sender by reply email, with a copy to ITSecurity@susquehannahealth.org < mailto:ITSecurity@susquehannahealth.org>, and then promptly delete the original and reply messages. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Mon May 4 11:26:38 2009 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon May 4 11:26:46 2009 Subject: [Histonet] Destain Hematoxylon In-Reply-To: <49FEB612.4060200@vneubert.com> Message-ID: <4EBFF65383B74D49995298C4976D1D5E03835CBC@LSRIEXCH1.lsmaster.lifespan.org> Simplest way is with 1% HCl in 70% ethanol. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of V. Neubert > Sent: Monday, May 4, 2009 5:32 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Destain Hematoxylon > > Hi, > > I want to destain hematoxylin (Shandon's Gill II). How to do that? > > Thanks in advance, > > V. Neubert > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From srishan <@t> mail.holyname.org Mon May 4 13:43:05 2009 From: srishan <@t> mail.holyname.org (srishan@mail.holyname.org) Date: Mon May 4 13:43:34 2009 Subject: [Histonet] XXXL GLOVES Message-ID: Hi All, We have a morgue attendent who does not fit into the XXL gloves. Does anyone know a vendor who can provide XXXL GLOVES latex (powder free) prefered, but will take any other brand too. I do really appreciate the help! Thanks Nirmala Srishan Holy Name Hospital Teaneck NJ 07666 From matthewtclose <@t> gmail.com Mon May 4 14:03:13 2009 From: matthewtclose <@t> gmail.com (Matthew Close) Date: Mon May 4 14:03:23 2009 Subject: [Histonet] re: destain hematoxylin Message-ID: <6abc767b0905041203h3537a3e8l6c0c959248b27c07@mail.gmail.com> 1) Acid alcohol: I use several drops of concentrated HCL in 95% ethanol (1% HCL in 70% alcohol well too). Simply dip your rinsed slides (or sections) into solution a couple of times, rinse and check them... a dip in water with about 10drops of ammonia should follow as well... 2) Saturated picric acid, followed by a couple of dips in water with several drops of ammonia, rinse and check 3) 2% Iron alum or ferric chloride (times vary and can range from a couple of dips to several minutes...) -Matt Close From janderson <@t> halozyme.com Mon May 4 14:04:57 2009 From: janderson <@t> halozyme.com (Jennifer Anderson) Date: Mon May 4 14:05:07 2009 Subject: [Histonet] direct-conjugates for IHC Message-ID: Hello everyone! I work with a scientist who insists on using primary antibodies directly conjugated with FITC or Texas Red for IHC. In my past experience these directly conjugated antibodies didn't give a strong enough signal for use in IHC (I've seen them used only for FACS analysis). I would appreciate your professional comments. Thanks again, so much, for your insight! Jennifer M. Anderson, Scientist Halozyme Therapeutics, Inc. 11404 Sorrento Valley Road San Diego, CA 92121 858-704-8333 janderson@halozyme.com The information transmitted in this email is confidential and is intended only for the person(s) or entity to which it is addressed. Delivery of this message to any person other than the intended recipient(s) is not intended in any way to waive confidentiality or any applicable privilege. Any review, retransmission, dissemination or other use of, or taking of any action in reliance upon, this information by individuals or entities other than the intended recipient is prohibited by Halozyme and may be in violation of applicable laws. If you received this in error, please contact the sender and delete/destroy this email. From shive003 <@t> umn.edu Mon May 4 14:41:27 2009 From: shive003 <@t> umn.edu (Jan Shivers) Date: Mon May 4 14:41:35 2009 Subject: [Histonet] (canine) eosinophil IHC Message-ID: Hello all, I'm asking this question on behalf of a researcher here. Does anyone know of the availability of an antibody that reacts with eosinophils, and if so, do you know if it reacts with canine eosinophils? Thank you in advance. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu From rjbuesa <@t> yahoo.com Mon May 4 14:52:59 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon May 4 14:53:03 2009 Subject: [Histonet] (canine) eosinophil IHC In-Reply-To: Message-ID: <700265.90361.qm@web65704.mail.ac4.yahoo.com> Any good H&E stain will reveal the eosinophils. Ren? J. --- On Mon, 5/4/09, Jan Shivers wrote: From: Jan Shivers Subject: [Histonet] (canine) eosinophil IHC To: "histonet" Date: Monday, May 4, 2009, 3:41 PM Hello all, I'm asking this question on behalf of a researcher here. Does anyone know of the availability of an antibody that reacts with eosinophils, and if so, do you know if it reacts with canine eosinophils? Thank you in advance. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histonetalias <@t> gmail.com Mon May 4 16:47:13 2009 From: histonetalias <@t> gmail.com (Histonet Alias) Date: Mon May 4 16:47:19 2009 Subject: [Histonet] Destain Hematoxylon In-Reply-To: <49FEB612.4060200@vneubert.com> References: <49FEB612.4060200@vneubert.com> Message-ID: <4b6c85510905041447g12779344t9dc8e21bc8ec356c@mail.gmail.com> Acid alcohol. It may not remove all of it but it will remove the majority. On Mon, May 4, 2009 at 5:32 AM, V. Neubert wrote: > Hi, > > I want to destain hematoxylin (Shandon's Gill II). How to do that? > > Thanks in advance, > > V. Neubert > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Al Ias HT(ASCP) Histology Manager Pathology Laboratory United States From derric57 <@t> vetmed.wsu.edu Mon May 4 19:53:16 2009 From: derric57 <@t> vetmed.wsu.edu (Phillips, Derrick) Date: Mon May 4 19:53:24 2009 Subject: [Histonet] Questions regarding tissue shrinkage during fixation and decalcification Message-ID: <642955D768EFCA46BA15AF5CFDA77F051D7B16610F@CVMMBX.vetmed.wsu.edu> Dear Histonet, I am new to histonet, and relatively new to histology. I have been digging through the histonet archives and doing searches on pubmed to try and answer some questions to a few problems I've been having. It has been tough finding a definitive answer to my questions so I am writing my first email, hoping someone out there can help me out. I'm trying to section through the the head of an adult rat. We've placed a recording electrode between the skull and the dura, and we are trying to section through all of the tissue to see how successful we are in electrode placement. The first problem I have been having is the amount the brain is shrinking (at least I believe it is due to the brain shrinking). I perfuse first with 0.9% saline, then fix with 10% formalin in PBS. Then it sits for 2 days in 10% formalin PBS + 20% sucrose. Afterward it is decalcified in 5% formic acid in DI for 4-5 days then in 5% gelatin for 24 hours, followed by 10% gelatin, then placed in the fridge to harden. After this it goes into -80, cut down, then sectioned in a cryostat. >From what I understand, fixing in formalin reduces the size of the brain tissue by about 20%. I've read that during the perfusion, instead of a prewash in 0.9% saline, using 9.45% sucrose helps to maintain the extracellular space and reduce the amount of shrinking. (Brain extracellular space fixed for electron microscopy. B. Cragg). I am wondering if anyone else has used this protocol and been successful. There was a post in the histonet archives from a Matt McElwee asking about a decal procedure that doesn't shrink or alter the brain, but the reply I found said that it wasn't necessary for the experiment he was doing. But it cited a protocol for decal by Gayle, but I was unable to locate this protocol. Is it possible that I am getting additional shrinkage from the decal process in addition to my fixation? If so, is there a way to reduce this? Another issue is the acceptance of the tissue to a glass slide. The brain and muscle tissue seem to take to the glass slides well, but the problem I've been having is with the bone. It tends to bow out and not adhere to the slide. This can be quite frustrating. I've tried charged slides, poly-l-lysine coated, silane coated, and superfrost plus slides, with minimal luck. Is there anything that can be done to help keep the bone on the slides? Finally, we are wanting to do triple fluorescent labelling of the tissue, the antigens need to remain intact. We are looking for proteins, so if something were to happen to the RNA, or DNA I wouldn't be as concerned. The search for answers on these topics has been frustrating at best. There seems to be a lot of variation in technique and I am unsure which will work the best for my needs. Any advice would be greatly appreciated!! Thank you! Derrick Phillips derric57@vetmed.wsu.edu From tifei <@t> foxmail.com Mon May 4 22:34:44 2009 From: tifei <@t> foxmail.com (TF) Date: Mon May 4 22:35:23 2009 Subject: [Histonet] Questions regarding tissue shrinkage during fixation anddecalcification References: <642955D768EFCA46BA15AF5CFDA77F051D7B16610F@CVMMBX.vetmed.wsu.edu> Message-ID: <200905051134389196202@foxmail.com> SGksIGEgMTk4NiBwYXBlciBpbiBKIEhpc3RvY2hlbSBDeXRvY2hlbSBpcyB2ZXJ5IHVzZWZ1bCB0 YWxraW5nIGFib3V0IGZpeGF0aW9uIHdpdGggZm9ybWFsaW4uDQppdCBpcyBzbyBnb29kIHRoYXQg YWZ0ZXIgSSByZWFkIGl0LCBJIGZvdW5kIHRoYXQgSSBoYXZlIGFza2VkIG1hbnkgInN0dXBpZCIg cXVlc3Rpb25zIGJlZm9yZS4NClRoZXkgZ2F2ZSBkZXRhaWxlZCBzaHJpbmthZ2UgY3VydmUgd2l0 aCBmaXhhdGl2ZSBjb25jZW50cmF0aW9uLCB0aXNzdWUgc2hyaW5rYWdlLg0KSW4geW91ciBjYXNl LCBpZiB5b3UgZml4IHRoZSBicmFpbiBhdCByb29tIHRlbXBlcmF0dXJlLCB0aGUgc2hyaW5rYWdl IGNhbiBiZSBuZWdsZWN0ZWQuDQoNCg0KMjAwOS0wNS0wNSANCg0KDQoNClRGIA0KDQoNCg0Kt6K8 /sjLo7ogUGhpbGxpcHMsIERlcnJpY2sgDQq3osvNyrG85KO6IDIwMDktMDUtMDUgIDA4OjU3OjI2 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ZHZpY2Ugd291bGQgYmUgZ3JlYXRseSBhcHByZWNpYXRlZCEhICBUaGFuayB5b3UhDQpEZXJyaWNr IFBoaWxsaXBzDQpkZXJyaWM1N0B2ZXRtZWQud3N1LmVkdQ0KX19fX19fX19fX19fX19fX19fX19f X19fX19fX19fX19fX19fX19fX19fX19fX18NCkhpc3RvbmV0IG1haWxpbmcgbGlzdA0KSGlzdG9u ZXRAbGlzdHMudXRzb3V0aHdlc3Rlcm4uZWR1DQpodHRwOi8vbGlzdHMudXRzb3V0aHdlc3Rlcm4u ZWR1L21haWxtYW4vbGlzdGluZm8vaGlzdG9uZXQNCg== From tifei <@t> foxmail.com Mon May 4 22:40:08 2009 From: tifei <@t> foxmail.com (TF) Date: Mon May 4 22:41:10 2009 Subject: [Histonet] 4% PFA/10% formalin in PB or in PBS? Message-ID: <200905051140032261926@foxmail.com> Hi, I want to ask this questions because I read different recipes in preparaing fixative solution... Here we perfuse animals with 0.9% saline, then 4% PFA in 0.1M PB...that is not a physiological solution... I see other people using 4% PFA in PBS.... will this lead to different brain shrinkage? Thanks. 2009-05-05 TF From koellingr <@t> comcast.net Mon May 4 22:54:41 2009 From: koellingr <@t> comcast.net (koellingr@comcast.net) Date: Mon May 4 22:54:44 2009 Subject: [Histonet] direct-conjugates for IHC In-Reply-To: Message-ID: <2091869083.2800171241495681048.JavaMail.root@sz0001a.emeryville.ca.mail.comcast.net> Jennifer, In the past, upon occasion when need arose, I'd use directly conjugated primary antibodies.? Mainly biotin or dig or peroxidase but also FITC directly to primary.? Then can look at it fluorescently or come back with an anti-FITC of some kind (several are good). Used these for my flow but also a lot of IHC.? Have never tried the Texas Red route for IHC.? What you say is true; direct conjugation can end up giving weaker signal but it doesn't have to be so bad.? 1) If you are doing the conjugation, that is great and you have a chance to play with the system.? Conjugation can result in near loss of antibody to bind or can result in good Ab binding or if you have the knack it can result in little loss of binding.? For flow, the conjugation is not critical since as long as you don't ruin the antibody, it will work since flow reagents are generally used in excess since there is no background (the background is plain sheath fluid) although certainly you can put in too much.? For IHC you need to be gentler in conjugation.? Don't know your company? but if a biotech you might have a BiaCore machine? (measure affinity of Ab's) or there are other ways to measure affinity.? So I would always keep an aliquot of native antibody and after I did my conjugation, give those people both samples to measure affinity (or how much loss).? Tell me immediately if I lost too much Ab binding ability because of my (hopefully not poor) conjugation.? In general yes, directly conjugating your primary can weaken signal but it doesn't have to be as much as conventional wisdom or lore makes it. Raymond Koelling PhenoPath Labs Seattle, WA ----- Original Message ----- From: "Jennifer Anderson" To: histonet@lists.utsouthwestern.edu Sent: Monday, May 4, 2009 12:04:57 PM GMT -08:00 US/Canada Pacific Subject: [Histonet] direct-conjugates for IHC Hello everyone! ? I work with a scientist who insists on using primary antibodies directly conjugated with FITC or Texas Red for IHC. ?In my past experience these directly conjugated antibodies didn't give a strong enough signal for use in IHC (I've seen them used only for FACS analysis). ?I would appreciate your professional comments. Thanks again, so much, for your insight! ? Jennifer M. Anderson, Scientist Halozyme Therapeutics, Inc. 11404 Sorrento Valley Road San Diego, CA 92121 858-704-8333 janderson@halozyme.com ? The information transmitted in this email is confidential and is intended only for the person(s) or entity to which it is addressed. ?Delivery of this message to any person other than the intended recipient(s) is not intended in any way to waive confidentiality or any applicable privilege. ?Any review, retransmission, dissemination or other use of, or taking of any action in reliance upon, this information by individuals or entities other than the intended recipient is prohibited by Halozyme and may be in violation of applicable laws. ?If you received this in error, please contact the sender and delete/destroy this email. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> waht.swest.nhs.uk Tue May 5 05:54:10 2009 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue May 5 05:54:20 2009 Subject: [Histonet] Destain Hematoxylon Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06A2A384@wahtntex2.waht.swest.nhs.uk> >From memory I found that it was better to give a good water wash or go into bicarb after destaining with acid/ alcohol as the acidity impacted on the staining if you went from acid/ alcohol directly into haematoxylin again (if you wanted to restain it). Kemlo Rogerson e-mail kemlorogerson@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histonet Alias Sent: 04 May 2009 22:47 To: V. Neubert Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Destain Hematoxylon Acid alcohol. It may not remove all of it but it will remove the majority. On Mon, May 4, 2009 at 5:32 AM, V. Neubert wrote: > Hi, > > I want to destain hematoxylin (Shandon's Gill II). How to do that? > > Thanks in advance, > > V. Neubert > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Al Ias HT(ASCP) Histology Manager Pathology Laboratory United States _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> waht.swest.nhs.uk Tue May 5 05:55:39 2009 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue May 5 05:55:43 2009 Subject: [Histonet] (canine) eosinophil IHC Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06A2A385@wahtntex2.waht.swest.nhs.uk> As already stated a good H&E will, but a bad one will show them even better; wash most of the E out of the H&E and the eosiniphils tend to let go of it last. Kemlo Rogerson e-mail kemlorogerson@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: 04 May 2009 20:41 To: histonet Subject: [Histonet] (canine) eosinophil IHC Hello all, I'm asking this question on behalf of a researcher here. Does anyone know of the availability of an antibody that reacts with eosinophils, and if so, do you know if it reacts with canine eosinophils? Thank you in advance. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brett_connolly <@t> merck.com Tue May 5 09:19:31 2009 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Tue May 5 09:19:44 2009 Subject: [Histonet] Stain for iodine? Message-ID: <63EA0607835FBA4689CEA9EA8B48269201EBB9FB@usctmx1141.merck.com> Histonetters, We have tissue that contains a iodinated nanosphere CT contrast agent and would like to identify the cells that took up said agent. Other than doing TEM does anyone have a suggestion for a stain we could use to visualize the iodine? Can be for either frozen or paraffin sections. Thanks, Brett Brett M. Connolly, Ph.D. Research Fellow, Imaging Dept. Merck & Co., Inc. PO Box 4, WP-44K West Point, PA 19486 tel. 215-652-2501 fax. 215-993-6803 brett_connolly@merck.com Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu - direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. From mcauliff <@t> umdnj.edu Tue May 5 09:26:36 2009 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Tue May 5 09:27:09 2009 Subject: [Histonet] Questions regarding tissue shrinkage during fixation and decalcification In-Reply-To: <642955D768EFCA46BA15AF5CFDA77F051D7B16610F@CVMMBX.vetmed.wsu.edu> References: <642955D768EFCA46BA15AF5CFDA77F051D7B16610F@CVMMBX.vetmed.wsu.edu> Message-ID: <4A004C9C.2080405@umdnj.edu> Hello Derrik: To get a good idea of how fixatives affect shrinkage go to the library (yes, the library, this info will not be online) and get a copy of "Principles of Biological Microtechnique" by J. R. Baker. That said, I suggest fixing for 5-7 days, 48 hours is a minimum. Formalin fixes tissue very slowly. Once the brain is nice and firm take it out of the skull and solve your decal problems. Or cut the head so you only have to decal a 1 cm slice and shorten the decal time. Also, is formic acid the best for skull? Just because someone else uses formic acid does not mean it is appropriate for your application. People have been using histology to see the exact location of electrodes for a long time, 60 years at least. Your problem has been solved before (more than once) and, since the older literature is not online a trip to the library is a MUST. Geoff Phillips, Derrick wrote: > Dear Histonet, > > > > I am new to histonet, and relatively new to histology. I have been digging through the histonet archives and doing searches on pubmed to try and answer some questions to a few problems I've been having. It has been tough finding a definitive answer to my questions so I am writing my first email, hoping someone out there can help me out. > > > > I'm trying to section through the the head of an adult rat. We've placed a recording electrode between the skull and the dura, and we are trying to section through all of the tissue to see how successful we are in electrode placement. The first problem I have been having is the amount the brain is shrinking (at least I believe it is due to the brain shrinking). > > > > I perfuse first with 0.9% saline, then fix with 10% formalin in PBS. > > Then it sits for 2 days in 10% formalin PBS + 20% sucrose. > > Afterward it is decalcified in 5% formic acid in DI for 4-5 days > > then in 5% gelatin for 24 hours, followed by 10% gelatin, then placed in the fridge to harden. After this it goes into -80, cut down, then sectioned in a cryostat. > > > > >From what I understand, fixing in formalin reduces the size of the brain tissue by about 20%. I've read that during the perfusion, instead of a prewash in 0.9% saline, using 9.45% sucrose helps to maintain the extracellular space and reduce the amount of shrinking. (Brain extracellular space fixed for electron microscopy. B. Cragg). I am wondering if anyone else has used this protocol and been successful. > > > > There was a post in the histonet archives from a Matt McElwee asking about a decal procedure that doesn't shrink or alter the brain, but the reply I found said that it wasn't necessary for the experiment he was doing. But it cited a protocol for decal by Gayle, but I was unable to locate this protocol. Is it possible that I am getting additional shrinkage from the decal process in addition to my fixation? If so, is there a way to reduce this? > > > > Another issue is the acceptance of the tissue to a glass slide. The brain and muscle tissue seem to take to the glass slides well, but the problem I've been having is with the bone. It tends to bow out and not adhere to the slide. This can be quite frustrating. I've tried charged slides, poly-l-lysine coated, silane coated, and superfrost plus slides, with minimal luck. Is there anything that can be done to help keep the bone on the slides? > > > > Finally, we are wanting to do triple fluorescent labelling of the tissue, the antigens need to remain intact. We are looking for proteins, so if something were to happen to the RNA, or DNA I wouldn't be as concerned. > > > > The search for answers on these topics has been frustrating at best. There seems to be a lot of variation in technique and I am unsure which will work the best for my needs. Any advice would be greatly appreciated!! Thank you! > > > > Derrick Phillips > > derric57@vetmed.wsu.edu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcauliff@umdnj.edu ********************************************** From relia1 <@t> earthlink.net Tue May 5 09:44:48 2009 From: relia1 <@t> earthlink.net (Pam Barker) Date: Tue May 5 09:44:53 2009 Subject: [Histonet] RELIA Histology Careers Bulletin 5/5/2009 Message-ID: Hi Histonetters!! It's Cinco De Mayo, Do me a favor and put that margarita down for a second and take a look at some of the opportunities I am working on. I have some great positions with some excellent employers. They are full time permanent positions with competitive wages and benefits and relocation assistance. Here are my management positions: Histology Supervisor - Bedford, MA Histology Supervisor - Los Angeles Here are my histotech positions: Lead Histotechnologist - Peabody, MA Lead Histotechnologist - Springfield, MA Histotechnician/Histotechnologist - Waco, TX Histotechnician/Histotechnologist - Corpus Christi, TX If you or anyone you know might be interested in these opportunties please contact me at relia1@earthlink.net or toll free at 866-607-3542. If you are on twitter you can find me at PamatRELIA. Have a great day!! Thank You! Pam Barker President RELIA Solutions Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 Toll Free: (866)607-3542 FAX: (407)678-2788 E-mail: relia1@earthlink.net <> www.myspace.com/pamatrelia From hodges420 <@t> msn.com Tue May 5 10:25:43 2009 From: hodges420 <@t> msn.com (MARY T HODGES) Date: Tue May 5 10:25:47 2009 Subject: [Histonet] need tow histo teck in Tucson Az Message-ID: Hi all . Is any one looking for a job in Tucson Az, as a histotechologist please reply if interested. Tere Hodges St Mary's Hospital _________________________________________________________________ Insert movie times and more without leaving Hotmail?. http://windowslive.com/Tutorial/Hotmail/QuickAdd?ocid=TXT_TAGLM_WL_HM_Tutorial_QuickAdd1_052009 From rjbuesa <@t> yahoo.com Tue May 5 11:03:41 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue May 5 11:03:45 2009 Subject: [Histonet] Stain for iodine? In-Reply-To: <63EA0607835FBA4689CEA9EA8B48269201EBB9FB@usctmx1141.merck.com> Message-ID: <919797.28706.qm@web65715.mail.ac4.yahoo.com> As you point out TEM would be?one way. Perhaps Frozen sections could allow you to visualize iodine crystals, but completely rule out the paraffin way because the iodine will dissolve during processing. Ren? J. --- On Tue, 5/5/09, Connolly, Brett M wrote: From: Connolly, Brett M Subject: [Histonet] Stain for iodine? To: histonet@lists.utsouthwestern.edu Date: Tuesday, May 5, 2009, 10:19 AM Histonetters, We have tissue that contains a iodinated nanosphere CT contrast agent and would like to identify the cells that took up said agent. Other than doing TEM does anyone have a suggestion for a stain we could use to visualize the iodine? Can be for either frozen or paraffin sections. Thanks, Brett Brett M. Connolly, Ph.D. Research Fellow, Imaging Dept. Merck & Co., Inc. PO Box 4, WP-44K West Point, PA 19486 tel. 215-652-2501 fax. 215-993-6803 brett_connolly@merck.com Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu - direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cbarone <@t> NEMOURS.ORG Tue May 5 11:53:46 2009 From: cbarone <@t> NEMOURS.ORG (Barone, Carol ) Date: Tue May 5 11:53:48 2009 Subject: [Histonet] Re:One -day meeting in VA, May 16th Message-ID: <37E4BAC017F57141AF64FAA5AEB04CE8033A7254@wlmmsx01.nemours.org> What? One day/2 seminars/ 5 CEU's.... in relevant area of IHC. When? May 16, 2009, Where? Old Domininion University Higher Education Center. Virginia Beach VA. Who? Hosted by the Regional states of DE, MD, NJ and PA...and supported by Biocare. For further information and registration: cbarone@nemours.org From lelmgren <@t> sunriselab.com Tue May 5 11:54:05 2009 From: lelmgren <@t> sunriselab.com (Laurie Elmgren) Date: Tue May 5 11:54:14 2009 Subject: [Histonet] Saturday coverage Message-ID: <4A672C6AE0402D4A89ECE29E8A4B47E3010F4A6E@MailPDC.sunriselab.com> Hi Hanita, Haiedeh will be available to work this Saturday. Laurie Laurie Elmgren Histology Supervisor Sunrise Medical Labs 240 Motor Pkwy Hauppauge, NY 11788 (631)435-1515x1108 "This message contains privileged and confidential information intended only for the use of the addressee named above. If you are not the intended recipient of this message you must not disseminate, copy or take any action in reliance on it." From Paul.Tharp <@t> onassignment.com Tue May 5 12:03:50 2009 From: Paul.Tharp <@t> onassignment.com (Paul Tharp) Date: Tue May 5 12:03:54 2009 Subject: [Histonet] Temporary Histo Tech position - Minneapolis/St. Paul, MN Message-ID: <55F6BEB2C20EE348A8DBE5AD26CDD77204687930@OASCINEX01.oaifield.onasgn.com> Hi all. We are currently hiring for a part time Histo Tech position in the Twin Cities Metro area and was curious if anyone is interested in hearing about this or knows someone who might be interested in this. The candidate would have to be local...no travelers for this one. Please let me know if anyone is interested in hearing more details...I look forward to hearing from you. Thanks! Paul Tharp Account Executive - St. Paul, MN On Assignment Healthcare Staffing t: 651.647.1160 x12009 c: 651.212.3699 f: 866.670.0731 toll free: 800.279.2345 paul.tharp@onassignment.com www.oahealthcare.com NASDAQ: ASGN People First. From JonSorenson <@t> chiwest.com Tue May 5 12:07:17 2009 From: JonSorenson <@t> chiwest.com (Sorenson, Jon (Nampa)) Date: Tue May 5 12:07:28 2009 Subject: [Histonet] RE: Histonet] xylene References: <20090502170616.5227A16BNW@email3.catholichealth.net> Message-ID: Loretta, When I was using the CBG recycler at a former employer, we had a similar incident, because the drain was plugged when the waste container got too full and the paraffin solidified in the drain tube. Check that possibility. Jon Jon Sorenson Histology Coordinator Mercy Medical Center JonSorenson@chiwest.com 208-463-5267 Message: 1 Date: Tue, 28 Apr 2009 14:06:04 -0400 From: "Moffatt, Loretta" Subject: [Histonet] xylene To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" We have recently had a problem with poor tissue processing (soft to cut/ poor staining). None of the tissue was processed the first time, and ultimately, some fragile tissue did not survive. The alcohols checked out and the processor, itself, does not appear to be an issue. We used recycled xylene in the processor first, followed by fresh xylene. The xylene, which came from a CBG recycler, failed the test for contamination, after the fact. Has anyone seen this kind of problem? We would appreciate any input. Thank you. Loretta Moffatt, MT(ASCP),MHA Laboratory Manager 777 Rural Avenue Williamsport, PA 17701 570-321-2326 (F)570-321-2489 lmoffatt@susquehannahealth.org Patience is never more important than when you are on the verge of losing it. From whitmorel <@t> mindspring.com Tue May 5 12:28:13 2009 From: whitmorel <@t> mindspring.com (whitmorel) Date: Tue May 5 12:28:17 2009 Subject: [Histonet] Re: Histonet Digest, Vol 66, Issue 5 message 18 Histo meeting Message-ID: <29111946.1241544493497.JavaMail.root@elwamui-little.atl.sa.earthlink.net> If anyone is going to the Virginia Beach seminar from the Charlottesville area or passing through the area on the way to the meeting, would you like to car pool? If you would, please email me at whitmorel@mindspring.com. If you like to drive, I will be happy to help with gas money. Lynn Whitmore (ASCP) Advanced Dermatology of Charlottesville (434)977-0027 --------- > >Message: 18 >Date: Tue, 5 May 2009 12:53:46 -0400 >From: "Barone, Carol " >Subject: [Histonet] Re:One -day meeting in VA, May 16th >To: histonet@lists.utsouthwestern.edu >Message-ID: > <37E4BAC017F57141AF64FAA5AEB04CE8033A7254@wlmmsx01.nemours.org> >Content-Type: text/plain; charset=iso-8859-1 > >What? One day/2 seminars/ 5 CEU's.... in relevant area of IHC. When? May 16, 2009, Where? Old Domininion University Higher Education Center. Virginia Beach VA. >Who? Hosted by the Regional states of DE, MD, NJ and PA...and supported by Biocare. For further information and registration: cbarone@nemours.org > > >------------------------------ > > From akbitting <@t> geisinger.edu Tue May 5 13:06:48 2009 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Tue May 5 13:07:02 2009 Subject: [Histonet] lifting tissue from glass slides Message-ID: <4A0047F7.2B7F.00C9.0@geisinger.edu> Does anyone know of a way to take an old H&E slide and remove a section from it to transfer it to another slide? I thought there was a way to do it with tape, but I can't find any information on the web about it. All I could find was tape to transfer frozen sections. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 No trees were hurt in the sending of this email However many electrons were severly inconvienienced! IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Bitting, Angela TEL;WORK:570-271-6844 ORG:;Histology EMAIL;WORK;PREF;NGW:AKBITTING@geisinger.edu N:Bitting;Angela END:VCARD From akemiat3377 <@t> yahoo.com Tue May 5 13:09:01 2009 From: akemiat3377 <@t> yahoo.com (Akemi Allison-Tacha) Date: Tue May 5 13:09:04 2009 Subject: [Histonet] (no subject) Message-ID: <786761.18166.qm@web31301.mail.mud.yahoo.com> Hi All Histology Manager's, I would like to ask you for your assistance.? I started my new position at APMG Labs yesterday, and one of the 1st requests given to me was by the HR Manager.? She is in the process of updating and revising the Job Descriptions and the Annual Performance Appraisal Forms for all the Histology Staff.? The positions are for the following: Histology Manager Histology Coordinator Lead Histologist Histotechnician (Non-Registered) Histology Lab Assistant Histology QC/Lab Assistant Grossing Assistant For the Annual Performance Appraisal Forms, she would like to separate each positions tasks for evaluation.? If any of you would like to share your existing forms, I would greatly appreciate any and all assistance. Thank You in advance for your assistance, Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Ct. Los Gatos, CA 95032 Cell: 425.941.4287? E-Mail: aallison-tacha@apmg.com From akemiat3377 <@t> yahoo.com Tue May 5 13:13:07 2009 From: akemiat3377 <@t> yahoo.com (Akemi Allison-Tacha) Date: Tue May 5 13:13:11 2009 Subject: [Histonet] Job Descriptions and Annual Performance Appraisal Forms Message-ID: <502214.69864.qm@web31304.mail.mud.yahoo.com> Sorry, I forgot to put the "Subject" Hi All Histology Manager's, I would like to ask you for your assistance.? I started my new position at APMG Labs yesterday, and one of the 1st requests given to me was by the HR Manager.? She is in the process of updating and revising the Job Descriptions and the Annual Performance Appraisal Forms for all the Histology Staff.? The positions are for the following: Histology Manager Histology Coordinator Lead Histologist Histotechnician (Non-Registered) Histology Lab Assistant Histology QC/Lab Assistant Grossing Assistant For the Annual Performance Appraisal Forms, she would like to separate each positions tasks for evaluation.? If any of you would like to share your existing forms, I would greatly appreciate any and all assistance. Thank You in advance for your assistance, Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Ct. Los Gatos, CA 95032 Cell: 425.941.4287? E-Mail: aallison-tacha@apmg.com Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Ct. Los Gatos, CA 95032 Cell: 425.941.4287? E-Mail: aallison-tacha@apmg.com From jrobertson <@t> pathologysciences.com Tue May 5 13:49:34 2009 From: jrobertson <@t> pathologysciences.com (Jodie Robertson) Date: Tue May 5 13:49:38 2009 Subject: [Histonet] lifting tissue from glass slides In-Reply-To: <4A0047F7.2B7F.00C9.0@geisinger.edu> References: <4A0047F7.2B7F.00C9.0@geisinger.edu> Message-ID: <518CD6920AA7154193CBE5977CD880733A8999@psmgsrv2.PSMG.local> Contact Newcomer Supply. They carry Mount-Quick and it can be used as a tissue transfer medium on stained H&E slides. Jodie Robertson, HT (ASCP) QIHC Pathology Sciences Medical Group -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Tuesday, May 05, 2009 11:07 AM To: histonet@lists.utsouthwestern.edu; Histonet-requests@lists.utsouthwestern.edu Subject: [Histonet] lifting tissue from glass slides Does anyone know of a way to take an old H&E slide and remove a section from it to transfer it to another slide? I thought there was a way to do it with tape, but I can't find any information on the web about it. All I could find was tape to transfer frozen sections. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 No trees were hurt in the sending of this email However many electrons were severly inconvienienced! IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From cbarone <@t> NEMOURS.ORG Tue May 5 14:14:02 2009 From: cbarone <@t> NEMOURS.ORG (Barone, Carol ) Date: Tue May 5 14:13:57 2009 Subject: [Histonet] antibody Message-ID: <37E4BAC017F57141AF64FAA5AEB04CE8033A7255@wlmmsx01.nemours.org> jan, We have sucessfully used Human Eosonophil Major Basic Protein - clone BMK-13 on F.S. (G.I. tissue ) for both resting an activated eosinophils. This can be used as a pan-eosinophilic marker in humans..sorry no data on canine ... (from Monosan) cat# mon 6008. From janet.dertien <@t> ttuhsc.edu Tue May 5 14:56:31 2009 From: janet.dertien <@t> ttuhsc.edu (Dertien, Janet) Date: Tue May 5 14:56:37 2009 Subject: [Histonet] freezing microtome Message-ID: We are interested in purchasing a freezing microtome or a vibratome and would appreciate some recommendations. We will be sectioning fixed rat brains for immunohistochemistry. Thanks!! From joseph-galbraith <@t> uiowa.edu Tue May 5 15:28:01 2009 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Tue May 5 15:28:06 2009 Subject: [Histonet] Snap-Frost System Message-ID: Hello: Does anyone have any experience with using the Snap-Frost rapid freezing system from Alphelys (France)? Our application is predominantly for tissue banking though we do a booming business in intraoperative diagnosis as well. The company and one review article {Virchow Arch (2008) 452:305-312} claim good RNA/DNA recovery from OCT embedded, isopentane frozen tissue using the system. They also claim good to excellent morphology at -80 in isopentane compared to liquid nitrogen at -190 and especially compared to CO2 freezing. Can anyone confirm or refute the claims? Does anyone know of a US based supplier of the Snap-Frost system? Thanks. Joe Galbraith University of Iowa Tissue Repository From PMonfils <@t> Lifespan.org Tue May 5 15:42:40 2009 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Tue May 5 15:42:46 2009 Subject: [Histonet] Stain for iodine? In-Reply-To: <63EA0607835FBA4689CEA9EA8B48269201EBB9FB@usctmx1141.merck.com> Message-ID: <4EBFF65383B74D49995298C4976D1D5E03835CBE@LSRIEXCH1.lsmaster.lifespan.org> Just a suggestion. Haven't tried it. A classic method for demonstrating starch is exposure to iodine, resulting in a dark purple to black precipitate. I wonder if you could use the technique in reverse, so to speak, and stain the iodine by exposure to a starch solution?? From Leah.Nichols <@t> cls.ab.ca Tue May 5 15:54:51 2009 From: Leah.Nichols <@t> cls.ab.ca (Leah Nichols) Date: Tue May 5 15:54:56 2009 Subject: [Histonet] Just In Time Slide Labelling Process Message-ID: <9166D14F25C6FE418CAFA9845BE52DB92E35261717@EXMBXC2.crha.bewell.ca> Our lab is investigating a "Just -in-Time" slide labelling process, at the point where slides are created. Our MLAs are currently pre-labelling our slides the night before. I am looking for information regarding how this process is handled in other laboratories (when are slides labelled, hand written or printed label, done per case or per worksheet, etc.) Also, when do you label your cassettes for grossing. Is pre-labelling done or are they made when the grossing is done? Thanks, Leah Nichols ________________________________ This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. From brett_connolly <@t> merck.com Tue May 5 16:05:35 2009 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Tue May 5 16:05:47 2009 Subject: [Histonet] Stain for iodine? In-Reply-To: <4EBFF65383B74D49995298C4976D1D5E03835CBE@LSRIEXCH1.lsmaster.lifespan.org> References: <63EA0607835FBA4689CEA9EA8B48269201EBB9FB@usctmx1141.merck.com> <4EBFF65383B74D49995298C4976D1D5E03835CBE@LSRIEXCH1.lsmaster.lifespan.org> Message-ID: <63EA0607835FBA4689CEA9EA8B48269201EBBC1A@usctmx1141.merck.com> Right, Lugol's sol'n with iodine and potassium iodide. That might be worth tinkering with, thanks. I was also thinking of something along the lines of crystal violet, ala a gram stain. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Tuesday, May 05, 2009 4:43 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Stain for iodine? Just a suggestion. Haven't tried it. A classic method for demonstrating starch is exposure to iodine, resulting in a dark purple to black precipitate. I wonder if you could use the technique in reverse, so to speak, and stain the iodine by exposure to a starch solution?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu - direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. From LSebree <@t> uwhealth.org Tue May 5 16:40:02 2009 From: LSebree <@t> uwhealth.org (Sebree Linda A) Date: Tue May 5 16:40:06 2009 Subject: [Histonet] Control & pt on same slide Message-ID: <5998F3BDFF7AAC4091C7AE93A7A1A5892CDFB8@UWHC-MAIL01.uwhis.hosp.wisc.edu> Histonetters, As a cost savings measure, we are considering doing away with using "red box" control slides. We would still like to put our control tissue on the same slide as our patient tissue. These are my questions: are people doing this and if so are you putting the control tissue on the top or bottom of the slide and how do you indicate which tissue is patient and which is control? Thanks for your responses. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 From gmartin <@t> marshallmedical.org Tue May 5 16:47:25 2009 From: gmartin <@t> marshallmedical.org (Martin, Gary) Date: Tue May 5 16:47:30 2009 Subject: [Histonet] Just In Time Slide Labeling Process In-Reply-To: <9166D14F25C6FE418CAFA9845BE52DB92E35261717@EXMBXC2.crha.bewell.ca> References: <9166D14F25C6FE418CAFA9845BE52DB92E35261717@EXMBXC2.crha.bewell.ca> Message-ID: <6ED9D4252F278841A0593D3D788AF24C05340607@mailsvr.MARSHMED.local> We are a small lab and hand write our slides at the microtome. Gary -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leah Nichols Sent: Tuesday, May 05, 2009 1:55 PM To: histonet@lists.utsouthwestern.edu Cc: Leah Nichols Subject: [Histonet] Just In Time Slide Labelling Process Our lab is investigating a "Just -in-Time" slide labelling process, at the point where slides are created. Our MLAs are currently pre-labelling our slides the night before. I am looking for information regarding how this process is handled in other laboratories (when are slides labelled, hand written or printed label, done per case or per worksheet, etc.) Also, when do you label your cassettes for grossing. Is pre-labelling done or are they made when the grossing is done? Thanks, Leah Nichols ________________________________ This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Tue May 5 17:17:39 2009 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Tue May 5 17:17:43 2009 Subject: [Histonet] Control & pt on same slide Message-ID: <57BE698966D5C54EAE8612E8941D76830580B714@EXCHANGE3.huntingtonhospital.com> We have never used the red box control slides. We put our control tissue at the top of the slide and the patient at the bottom. After the slide is coverslipped, we draw a line with an extra fine sharpie between the two and write "control" next to the control tissue. Laurie Colbert -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sebree Linda A Sent: Tuesday, May 05, 2009 2:40 PM To: Histonet Subject: [Histonet] Control & pt on same slide Histonetters, As a cost savings measure, we are considering doing away with using "red box" control slides. We would still like to put our control tissue on the same slide as our patient tissue. These are my questions: are people doing this and if so are you putting the control tissue on the top or bottom of the slide and how do you indicate which tissue is patient and which is control? Thanks for your responses. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From chana.de.wolf <@t> gmail.com Tue May 5 17:50:50 2009 From: chana.de.wolf <@t> gmail.com (Chana de Wolf) Date: Tue May 5 17:50:54 2009 Subject: [Histonet] Fixation for EM Message-ID: <3f4b71f10905051550j67ed2e0bl1a92343b63b3f042@mail.gmail.com> Hello all, I am new to Histonet and very happy to be here! Of course, I was brought here the same way most are -- I have questions! I am currently working on a project for a mathematician who wants to develop an algorithm that can be used to calculate length of ischemia when fed EM images of ischemic brains. In order to develop the algorithm, we must first generate EM images of brains after various periods of ischemia. Like most histologists, any brain fixations I have done have involved immediate perfusion fixation to *minimize* ischemia, so this is new and interesting territory. Based on my own experiences and a thorough reading of the literature, I assume that perfusion fixation of ischemic brains is not practical due to the "no-reflow" phenomenon. My proposal, then, is to hemisect and diffusion fix the brains. However, *I* am not performing the EMs, and I am not totally certain when the EMs will take place following fixation. My questions follow: (1) What is the best fixative solution to be used under the circumstances? I was thinking of using a combination of paraformaldehyde and acrolein due to acrolein's penetrative qualities and superior fixative strength. (2) Do I need to perform a secondary fix with osmium tetroxide myself...or do I leave that to the EM lab techs to perform when they make slices? If at all possible, I want to minimize the amount of tissue processing on my end. (3) How long can the brains be left in post-fixative solution prior to further processing in the EM lab? Or, in other words, is there a maximum time period after which fixed whole brains cannot be sliced, washed, embedded, or otherwise processed? (4) Any other suggestions, comments, etc.? Obviously, there has to be a less-than-ideal division of labor in this matter because we do not expect the EM lab to remove brains from rats that have been dead for up to 48 hours. I can handle that part, but I want to ensure I do everything right because what I do affects the rest of this study. Thank you for your time and any valuable insights into this matter. Chana de Wolf Advanced Neural Biosciences, Inc. From akemiat3377 <@t> yahoo.com Tue May 5 18:02:08 2009 From: akemiat3377 <@t> yahoo.com (Akemi Allison-Tacha) Date: Tue May 5 18:02:11 2009 Subject: [Histonet] Wrong work e-mail address for Job Discriptions Message-ID: <371561.17244.qm@web31308.mail.mud.yahoo.com> Hi All, Sorry, My Bad!? For those of you who were kind enough to supply me with information regarding Job Descriptions and Performance Appraisal Forms to my work e-mail address.? Guess I made a mistake on my new work e-mail address. Below is my correct e-mail address.? I would appreciate any information resent to either my personal or work e-mail. Thanks a bunch! Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Ct. Los Gatos, CA 95032 Cell: 425.941.4287 W: E-Mail: aallison-tacha@apmglab.com P: E-Mail: akemiat3377@yahoo.com From saby_joseph_a <@t> yahoo.com Tue May 5 18:39:28 2009 From: saby_joseph_a <@t> yahoo.com (Joseph Saby) Date: Tue May 5 18:39:32 2009 Subject: [Histonet] Staining Controls Message-ID: <84470.31338.qm@web33804.mail.mud.yahoo.com> Hello to Histoland- I have a question concerning staining controls. I am currently working for a GLP/GMP lab.? Their?use of staining controls requires the purchase of said controls from sources such as Histology Controls Systems (TM) where the manufacturer certifies that these controls are effective controls for the stain for which they are listed. Now, call me old school, but it has always seemed to me that many controls have internal control structures.? For instance, it seems superfluous to have a myelin control for a Luxol Fast Blue/PAS stain.? Or perhaps muscle (cardiac or skeletal) for a Masson's Trichrome.? Arterial wall for a Verheoff's Elastin stain.? Etc. Where I have worked in the past, we were always looking for excellent examples of tissue samples which exemplified excellent staining for control slides for?specifc stains.? My manager does not have a background in histology or pathology, and this concept is new to her.? Since she keeps a sharp eye on the bottom line, and staining control slides are ridiculously expensive, she is open to the suggestion that our lab should use our own control slides.? However, she feels I should seek outside opinions/testimonials about the certification process for said "house" controls, especially as this process relates to a GLP/GMP facility. This question should be good for a healthy debate.? I look forward to the coming suggestions! Joe Saby, BA, HT(ASCP) Supervisor of Histology NAMSA, Northwood Ohio From chana.de.wolf <@t> gmail.com Tue May 5 19:27:58 2009 From: chana.de.wolf <@t> gmail.com (Chana de Wolf) Date: Tue May 5 19:28:02 2009 Subject: [Histonet] Re: Fixation for EM In-Reply-To: <3f4b71f10905051550j67ed2e0bl1a92343b63b3f042@mail.gmail.com> References: <3f4b71f10905051550j67ed2e0bl1a92343b63b3f042@mail.gmail.com> Message-ID: <3f4b71f10905051727l6904edadu5fe9ab7412ad3dd3@mail.gmail.com> By the way, I should have mentioned that these are RAT brains I am talking about. Sorry! On Tue, May 5, 2009 at 3:50 PM, Chana de Wolf wrote: > Hello all, I am new to Histonet and very happy to be here! Of course, > I was brought here the same way most are -- I have questions! > > I am currently working on a project for a mathematician who wants to > develop an algorithm that can be used to calculate length of ischemia > when fed EM images of ischemic brains. In order to develop the > algorithm, we must first generate EM images of brains after various > periods of ischemia. Like most histologists, any brain fixations I > have done have involved immediate perfusion fixation to *minimize* > ischemia, so this is new and interesting territory. > > Based on my own experiences and a thorough reading of the literature, > I assume that perfusion fixation of ischemic brains is not practical > due to the "no-reflow" phenomenon. My proposal, then, is to hemisect > and diffusion fix the brains. However, *I* am not performing the EMs, > and I am not totally certain when the EMs will take place following > fixation. > > My questions follow: > > (1) What is the best fixative solution to be used under the > circumstances? I was thinking of using a combination of > paraformaldehyde and acrolein due to acrolein's penetrative qualities > and superior fixative strength. > > (2) Do I need to perform a secondary fix with osmium tetroxide > myself...or do I leave that to the EM lab techs to perform when they > make slices? If at all possible, I want to minimize the amount of > tissue processing on my end. > > (3) How long can the brains be left in post-fixative solution prior to > further processing in the EM lab? Or, in other words, is there a > maximum time period after which fixed whole brains cannot be sliced, > washed, embedded, or otherwise processed? > > (4) Any other suggestions, comments, etc.? Obviously, there has to be > a less-than-ideal division of labor in this matter because we do not > expect the EM lab to remove brains from rats that have been dead for > up to 48 hours. I can handle that part, but I want to ensure I do > everything right because what I do affects the rest of this study. > > Thank you for your time and any valuable insights into this matter. > > Chana de Wolf > Advanced Neural Biosciences, Inc. > From dcojita <@t> tampabay.rr.com Tue May 5 20:02:01 2009 From: dcojita <@t> tampabay.rr.com (dcojita@tampabay.rr.com) Date: Tue May 5 20:02:07 2009 Subject: [Histonet] Molecular testing ASCP In-Reply-To: <3f4b71f10905051727l6904edadu5fe9ab7412ad3dd3@mail.gmail.com> Message-ID: Hi Histonetters, Do any of you have a recommendation for reference books in order to prepare for the ASCP examine specializing in molecular? Any assistance you could provide would be greatly appreciated. From cfitz <@t> telus.net Tue May 5 20:14:27 2009 From: cfitz <@t> telus.net (Cathy) Date: Tue May 5 20:14:29 2009 Subject: [Histonet] Just In Time Slide Labelling Process In-Reply-To: <9166D14F25C6FE418CAFA9845BE52DB92E35261717@EXMBXC2.crha.bewell.ca> References: <9166D14F25C6FE418CAFA9845BE52DB92E35261717@EXMBXC2.crha.bewell.ca> Message-ID: Our MLA's labels the slides the day before from a worksheet that is generated from the gross room. The cassettes are made during the accessioning process and are then taken into the gross room; any additional cassettes that are needed are made by the PA. We have just been approved for a slides and cassette labeler. Cathy Fitzpatrick -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leah Nichols Sent: Tuesday, May 05, 2009 1:55 PM To: histonet@lists.utsouthwestern.edu Cc: Leah Nichols Subject: [Histonet] Just In Time Slide Labelling Process Our lab is investigating a "Just -in-Time" slide labelling process, at the point where slides are created. Our MLAs are currently pre-labelling our slides the night before. I am looking for information regarding how this process is handled in other laboratories (when are slides labelled, hand written or printed label, done per case or per worksheet, etc.) Also, when do you label your cassettes for grossing. Is pre-labelling done or are they made when the grossing is done? Thanks, Leah Nichols ________________________________ This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JEllin <@t> yumaregional.org Tue May 5 20:18:04 2009 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Tue May 5 20:18:12 2009 Subject: [Histonet] Just In Time Slide Labelling Process In-Reply-To: Message-ID: <29BE166A2CF48D459853F8EC57CD37E801A8323A@EXCHANGECLUSTER.yumaregional.local> We have done some extensive work on this subject, give me a call so that we can chat on this Jesus A Ellin HT/PA ASCP Department of Pathology/Histology Yuma Regional Medical Center 2400 South Ave A Yuma, AZ 85364 - 7170 ( Office: (928) 336-1743 ( Fax: (928) 336-7319 * Email: jellin@yumaregional.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cathy Sent: Tuesday, May 05, 2009 6:14 PM To: 'Leah Nichols'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Just In Time Slide Labelling Process Our MLA's labels the slides the day before from a worksheet that is generated from the gross room. The cassettes are made during the accessioning process and are then taken into the gross room; any additional cassettes that are needed are made by the PA. We have just been approved for a slides and cassette labeler. Cathy Fitzpatrick -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leah Nichols Sent: Tuesday, May 05, 2009 1:55 PM To: histonet@lists.utsouthwestern.edu Cc: Leah Nichols Subject: [Histonet] Just In Time Slide Labelling Process Our lab is investigating a "Just -in-Time" slide labelling process, at the point where slides are created. Our MLAs are currently pre-labelling our slides the night before. I am looking for information regarding how this process is handled in other laboratories (when are slides labelled, hand written or printed label, done per case or per worksheet, etc.) Also, when do you label your cassettes for grossing. Is pre-labelling done or are they made when the grossing is done? Thanks, Leah Nichols ________________________________ This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. 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 This message is confidential, intended only for the named recipient(s) and may contain information that is privileged or exempt from disclosure under applicable law. If you are not the intended recipient(s), you are notified that the dissemination, distribution, or copying of this message is strictly prohibited. If you receive this message in error, or are not the named recipient(s), please notify the sender at either the e-mail, fax, address, or telephone number listed above and delete this e-mail from your computer. Thank You. From Kemlo.Rogerson <@t> waht.swest.nhs.uk Wed May 6 02:41:21 2009 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Wed May 6 02:41:37 2009 Subject: [Histonet] Staining Controls Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06A2A462@wahtntex2.waht.swest.nhs.uk> You are of course correct; if the muscle layers haven't stained then the "muscle stain" hasn't worked. Unfortunately that is a common sensed approach, something that doesn't seem fashionable nowadays. I have said this before (concerning glycogen controls) but I'll say it again. If it is deemed that a control other than the internal be used then use pig organs or some other species. I guess if we look at the Chemistry and Haematology analogy then your need to determine precision (specificity) and sensitivity. You need to know that what you are staining is the substance in question and only the substance in question; does that make sense? In Blood Science they have 'internal controls' and external controls and it is the external control (NEQAS) that controls precision; that you are not only picking up the analyte but you are picking up the right amount compared to your peers. I suspect that's a way forward; use animal controls that are shown by your peer group to not only demonstrate the substance but that its being picked up in the correct quantities (its staining the same) and then use that. You can do that between a number of Labs thus cutting out the Retailer!! Bet the Retailer don't get agreement like that. Kemlo Rogerson e-mail kemlorogerson@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joseph Saby Sent: 06 May 2009 00:39 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Staining Controls Hello to Histoland- I have a question concerning staining controls. I am currently working for a GLP/GMP lab.? Their?use of staining controls requires the purchase of said controls from sources such as Histology Controls Systems (TM) where the manufacturer certifies that these controls are effective controls for the stain for which they are listed. Now, call me old school, but it has always seemed to me that many controls have internal control structures.? For instance, it seems superfluous to have a myelin control for a Luxol Fast Blue/PAS stain.? Or perhaps muscle (cardiac or skeletal) for a Masson's Trichrome.? Arterial wall for a Verheoff's Elastin stain.? Etc. Where I have worked in the past, we were always looking for excellent examples of tissue samples which exemplified excellent staining for control slides for?specifc stains.? My manager does not have a background in histology or pathology, and this concept is new to her.? Since she keeps a sharp eye on the bottom line, and staining control slides are ridiculously expensive, she is open to the suggestion that our lab should use our own control slides.? However, she feels I should seek outside opinions/testimonials about the certification process for said "house" controls, especially as this process relates to a GLP/GMP facility. This question should be good for a healthy debate.? I look forward to the coming suggestions! Joe Saby, BA, HT(ASCP) Supervisor of Histology NAMSA, Northwood Ohio _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ooi.ting.huay <@t> nhc.com.sg Wed May 6 07:29:00 2009 From: ooi.ting.huay <@t> nhc.com.sg (ooi.ting.huay@nhc.com.sg) Date: Wed May 6 07:30:30 2009 Subject: [Histonet] Different between MMA and PMMA embedded sample Message-ID: Hi, I would like to know what is the different between MMA and PM embedded sample? As I know, if we are dealing with MMA embedded sample, How about PMMA? Regards, Ooi From rjbuesa <@t> yahoo.com Wed May 6 07:42:38 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed May 6 07:42:46 2009 Subject: [Histonet] Just In Time Slide Labelling Process In-Reply-To: <9166D14F25C6FE418CAFA9845BE52DB92E35261717@EXMBXC2.crha.bewell.ca> Message-ID: <635088.15742.qm@web65704.mail.ac4.yahoo.com> Try contacting Dr. Richard Zarbo at the Henry Ford Hospital, he is using this method with great success. Ren? J. --- On Tue, 5/5/09, Leah Nichols wrote: From: Leah Nichols Subject: [Histonet] Just In Time Slide Labelling Process To: "histonet@lists.utsouthwestern.edu" Cc: "Leah Nichols" Date: Tuesday, May 5, 2009, 4:54 PM Our lab is investigating a "Just -in-Time" slide labelling process, at the point where slides are created. Our MLAs are currently pre-labelling our slides the night before. I am looking for information regarding how this process is handled in other laboratories (when are slides labelled, hand written or printed label, done per case or per worksheet, etc.) Also, when do you label your cassettes for grossing. Is pre-labelling done or are they made when the grossing is done? Thanks, Leah Nichols ________________________________ This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed May 6 07:46:44 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed May 6 07:46:53 2009 Subject: [Histonet] Control & pt on same slide In-Reply-To: <5998F3BDFF7AAC4091C7AE93A7A1A5892CDFB8@UWHC-MAIL01.uwhis.hosp.wisc.edu> Message-ID: <597909.95392.qm@web65710.mail.ac4.yahoo.com> Control always on the top of the slide (as if you were using the red square space) and you have to describe the change in your SOP, preferably with either a sketch or a photo. Ren? J. --- On Tue, 5/5/09, Sebree Linda A wrote: From: Sebree Linda A Subject: [Histonet] Control & pt on same slide To: "Histonet" Date: Tuesday, May 5, 2009, 5:40 PM Histonetters, As a cost savings measure, we are considering doing away with using "red box" control slides. We would still like to put our control tissue on the same slide as our patient tissue. These are my questions: are people doing this and if so are you putting the control tissue on the top or bottom of the slide and how do you indicate which tissue is patient and which is control? Thanks for your responses. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed May 6 07:53:53 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed May 6 07:54:01 2009 Subject: [Histonet] Staining Controls In-Reply-To: <84470.31338.qm@web33804.mail.mud.yahoo.com> Message-ID: <180942.54885.qm@web65713.mail.ac4.yahoo.com> You are completely right in what you have written. Internal controls are even better for IHC because the epitope signal is "normal" and better for the selection of dilution rather than using a pathological example that probably has a stronger signal and will determine a higher dilution. As to buying controls it is, for me, a waste of money. The control has to be selected and accepted by the pathologists, and that is more than enough. The thing is that, whatever you do, and however you do it, just write it in detail in your SOP and that, after being signed off by the lab director, is more than enough. Ren? J. --- On Tue, 5/5/09, Joseph Saby wrote: From: Joseph Saby Subject: [Histonet] Staining Controls To: histonet@lists.utsouthwestern.edu Date: Tuesday, May 5, 2009, 7:39 PM Hello to Histoland- I have a question concerning staining controls. I am currently working for a GLP/GMP lab.? Their?use of staining controls requires the purchase of said controls from sources such as Histology Controls Systems (TM) where the manufacturer certifies that these controls are effective controls for the stain for which they are listed. Now, call me old school, but it has always seemed to me that many controls have internal control structures.? For instance, it seems superfluous to have a myelin control for a Luxol Fast Blue/PAS stain.? Or perhaps muscle (cardiac or skeletal) for a Masson's Trichrome.? Arterial wall for a Verheoff's Elastin stain.? Etc. Where I have worked in the past, we were always looking for excellent examples of tissue samples which exemplified excellent staining for control slides for?specifc stains.? My manager does not have a background in histology or pathology, and this concept is new to her.? Since she keeps a sharp eye on the bottom line, and staining control slides are ridiculously expensive, she is open to the suggestion that our lab should use our own control slides.? However, she feels I should seek outside opinions/testimonials about the certification process for said "house" controls, especially as this process relates to a GLP/GMP facility. This question should be good for a healthy debate.? I look forward to the coming suggestions! Joe Saby, BA, HT(ASCP) Supervisor of Histology NAMSA, Northwood Ohio _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akbitting <@t> geisinger.edu Wed May 6 08:22:30 2009 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Wed May 6 08:22:42 2009 Subject: [Histonet] Gram & AFB staining FNA smears Message-ID: <4A0156D6.2B7F.00C9.0@geisinger.edu> The subject of staining FNA smears with AFB and Gram stains in Histology vs in Micro came up today. Is there a reason that FNAs can't be stained the same way we stain our tissue sections? One of our docs was under the impression that it's not acceptable to use the staining methods we use in our Histology lab. I don't know what method Micro labs use, so I was hoping someone could shed some light on this subject for me. Thanks, as always, Angie Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 No trees were hurt in the sending of this email However many electrons were severly inconvienienced! IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Bitting, Angela TEL;WORK:570-271-6844 ORG:;Histology EMAIL;WORK;PREF;NGW:AKBITTING@geisinger.edu N:Bitting;Angela END:VCARD From jrobertson <@t> pathologysciences.com Wed May 6 08:42:19 2009 From: jrobertson <@t> pathologysciences.com (Jodie Robertson) Date: Wed May 6 08:42:29 2009 Subject: [Histonet] Control & pt on same slide In-Reply-To: <5998F3BDFF7AAC4091C7AE93A7A1A5892CDFB8@UWHC-MAIL01.uwhis.hosp.wisc.edu> References: <5998F3BDFF7AAC4091C7AE93A7A1A5892CDFB8@UWHC-MAIL01.uwhis.hosp.wisc.edu> Message-ID: <518CD6920AA7154193CBE5977CD880733A899B@psmgsrv2.PSMG.local> We had to do away with the red control box slides due to having some staining issues with the Benchmark. One of our techs came up with a stamp that we stamp the back of our slides with Control on top, a line and patient on the bottom and we stamp it in the middle between the control and the patient on the back of the slide. The letters are reversed on the stamp so we can do this. It works really well and the pathologists are happy with this. It is better than handwriting. Jodie Robertson, HT (ASCP) QIHC Pathology Sciences Medical Group -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sebree Linda A Sent: Tuesday, May 05, 2009 2:40 PM To: Histonet Subject: [Histonet] Control & pt on same slide Histonetters, As a cost savings measure, we are considering doing away with using "red box" control slides. We would still like to put our control tissue on the same slide as our patient tissue. These are my questions: are people doing this and if so are you putting the control tissue on the top or bottom of the slide and how do you indicate which tissue is patient and which is control? Thanks for your responses. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tammy <@t> surgicalpathlabs.com Wed May 6 09:00:27 2009 From: tammy <@t> surgicalpathlabs.com (Tammy de Leon) Date: Wed May 6 09:12:33 2009 Subject: [Histonet] Florida Histo Openings Message-ID: Good morning. We are a Pathology Lab in Pinellas Park, Florida. We're looking to add to our team! Candidates should be an HTL or HT (ASCP) or equivalent and have a valid driver's license. Primary responsibilities include on site frozen sections including mobile laboratory units. We offer a full benefits package (SPL pays 100% of the employee premium for Medical, Dental, Life and LTD!!!) with a great working environment. View our Company Info and job details at www.surgicalpathlabs.com. Thanks. Tammy R. de Leon Surgical Pathology Laboratories Human Resources Officer 8455 66th Street N Pinellas Park, Fl 33781 Office Phone - 727-209-1215 Fax - 727-545-1644 From LSebree <@t> uwhealth.org Wed May 6 09:17:24 2009 From: LSebree <@t> uwhealth.org (Sebree Linda A) Date: Wed May 6 09:17:27 2009 Subject: [Histonet] Thanks for all the responses re: Control & pt on same slide Message-ID: <5998F3BDFF7AAC4091C7AE93A7A1A5892CDFBD@UWHC-MAIL01.uwhis.hosp.wisc.edu> Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 From jrobertson <@t> pathologysciences.com Wed May 6 09:20:00 2009 From: jrobertson <@t> pathologysciences.com (Jodie Robertson) Date: Wed May 6 09:20:08 2009 Subject: [Histonet] Control & pt on same slide In-Reply-To: <518CD6920AA7154193CBE5977CD880733A899E@psmgsrv2.PSMG.local> References: <518CD6920AA7154193CBE5977CD880733A899B@psmgsrv2.PSMG.local> <5998F3BDFF7AAC4091C7AE93A7A1A5892CDFBB@UWHC-MAIL01.uwhis.hosp.wisc.edu> <518CD6920AA7154193CBE5977CD880733A899D@psmgsrv2.PSMG.local> <518CD6920AA7154193CBE5977CD880733A899E@psmgsrv2.PSMG.local> Message-ID: <518CD6920AA7154193CBE5977CD880733A899F@psmgsrv2.PSMG.local> Sure. It looks like below except make the letters backwards. CONTROL PATIENT Our tech went to a business supply store in her home town and requested to have a stamp made. You can get a stamp made for anything. It was approximately $20.00 and she got a stamp pad and ink. We all talked about it for a while as we label the slides for the control pretty much as soon as we coverslip the slide. We coverslip by hand, so the coverslip isn't dry yet, therefore we decided it was easier to label the back of the slide. It's hard to write backwards for some, so we had some dyslexic moments. The stamp eliminated the dyslexia, it was neater and easier to read. Our pathologist wanted it labeled the complete word, not just drawing a line and using "C" and "P" to indicate like we tried to do. The ink doesn't come off unless it comes in contact with alcohol. If the stamp is smeared, we remove it with alcohol and repeat it. It dries right away and everyone seems happy with the results. If you have a tape automatic coverslipper, you could just stamp the front as they stay put, unlike a glass coverslip done by hand. We also called around to see if we could get coverslips made stamped with this and Erie Scientific would make it for us, but at a ridiculous cost as they thought it would compete with their red box slide or etched glass slide. Hope this helps. Jodie Robertson, HT (ASCP) QIHC Pathology Sciences Medical Group -----Original Message----- From: Sebree Linda A [mailto:LSebree@uwhealth.org] Sent: Wednesday, May 06, 2009 6:46 AM To: Jodie Robertson Subject: RE: [Histonet] Control & pt on same slide Very interesting. Can you tell me more about what type of stamp you had made? Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -----Original Message----- From: Jodie Robertson [mailto:jrobertson@pathologysciences.com] Sent: Wednesday, May 06, 2009 8:42 AM To: Sebree Linda A; Histonet Subject: RE: [Histonet] Control & pt on same slide We had to do away with the red control box slides due to having some staining issues with the Benchmark. One of our techs came up with a stamp that we stamp the back of our slides with Control on top, a line and patient on the bottom and we stamp it in the middle between the control and the patient on the back of the slide. The letters are reversed on the stamp so we can do this. It works really well and the pathologists are happy with this. It is better than handwriting. Jodie Robertson, HT (ASCP) QIHC Pathology Sciences Medical Group -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sebree Linda A Sent: Tuesday, May 05, 2009 2:40 PM To: Histonet Subject: [Histonet] Control & pt on same slide Histonetters, As a cost savings measure, we are considering doing away with using "red box" control slides. We would still like to put our control tissue on the same slide as our patient tissue. These are my questions: are people doing this and if so are you putting the control tissue on the top or bottom of the slide and how do you indicate which tissue is patient and which is control? Thanks for your responses. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed May 6 11:24:08 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed May 6 11:24:16 2009 Subject: [Histonet] Gram & AFB staining FNA smears In-Reply-To: <4A0156D6.2B7F.00C9.0@geisinger.edu> Message-ID: <609446.31160.qm@web65711.mail.ac4.yahoo.com> The "doc" is wrong, otherwise your histology sections to be stained with Gram & AFB should also be sent to micro. Perhaps you will not have to do them anymore. Your "doc's" reasoning is purely oxymoronic. Ren? J. --- On Wed, 5/6/09, Angela Bitting wrote: From: Angela Bitting Subject: [Histonet] Gram & AFB staining FNA smears To: histonet@lists.utsouthwestern.edu Date: Wednesday, May 6, 2009, 9:22 AM The subject of staining FNA smears with AFB and Gram stains in Histology vs in Micro came up today. Is there a reason that FNAs can't be stained the same way we stain our tissue sections? One of our docs was under the impression that it's not acceptable to use the staining methods we use in our Histology lab. I don't know what method Micro labs use, so I was hoping someone could shed some light on this subject for me. Thanks, as always, Angie Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 No trees were hurt in the sending of this email However many electrons were severly inconvienienced! IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you._______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vapatpxs <@t> yahoo.com Wed May 6 11:29:50 2009 From: vapatpxs <@t> yahoo.com (Va Paula Sicurello) Date: Wed May 6 11:30:00 2009 Subject: [Histonet] Fixation for EM Message-ID: <434684.38004.qm@web46113.mail.sp1.yahoo.com> Hi Chana, I'm going to highlight my answers after each of your questions below. Though I will say that you should contact whomever may be processing the brains for EM to see what they would prefer.? Also ask them which buffer they prefer.? Karnovsky's fix is traditionally made with Sodium cacodylate (an arsenic base compound) but some labs have switched to phosphate buffer for safety reasons. If you have any other questions, please feel free to contact me. Paula? :-) Paula Sicurello VA Medical Center San Diego Veterans Medical Research Foundation (VMRF) Core Research Imaging Center 3350 La Jolla Village Dr., MC151 San Diego, CA 92161 858-552-8585 x2397 http://www.vmrf.org/researchcenters/confocal/confocal.html > > My questions follow: > > (1) What is the best fixative solution to be used under > the > circumstances? I was thinking of using a combination of > paraformaldehyde and acrolein due to acrolein's penetrative > qualities > and superior fixative strength. ?I think most labs tend to stay away from acrolein (it is tear gas and not pleasant to be around).? A modified Karnovsky's is probably the best all around general EM fixative.? If it's OK to mince the brain into 1mm cubed bits that would be best for proper fixation.? If you can't then you end up with a bit of a fixation barrier.? Glutaraldehyde penetrates at 0.5mm/hour at room temp.? When the outside parts get fixed first and the sample is large, then fix for longer periods of time.? Samples can be left in the refrigerator for weeks. > > (2) Do I need to perform a secondary fix with osmium > tetroxide > myself...or do I leave that to the EM lab techs to perform > when they > make slices? If at all possible, I want to minimize the > amount of > tissue processing on my end. Unless you have to, I would avoid the Osmium textroxide.? A good EM lab would prefer that you just fix your samples and either submit the samples in buffer or fixative.? While acrolein is tear gas, Osmium textroxide is seriuosly poisonous.? Exposure to it can cause blindness and death.? With an exposure limit of 0.002ppm it is nasty.? The one good thing is you will go blind before you stop breathing and the blindness only lasts a few months.? So if you can't see or things get blurry, it's time to get some fresh air?? ;-).? ??? Seriously, let the EM people do the Osmication. > > (3) How long can the brains be left in post-fixative > solution prior to > further processing in the EM lab? Or, in other words, is > there a > maximum time period after which fixed whole brains cannot > be sliced, > washed, embedded, or otherwise processed? Theoretically, samples can be left in fix a long time.? I once forgot a monolayer of cells and left them in Karnovsky's for 2 weeks without any noticeable deterioration of the ultrastructure.? Again, ask the EM people, some have protocols they like to follow and times they prefer for optimization of image quality. > > (4) Any other suggestions, comments, etc.? Obviously, there > has to be > a less-than-ideal division of labor in this matter because > we do not > expect the EM lab to remove brains from rats that have been > dead for > up to 48 hours. I can handle that part, but I want to > ensure I do > everything right because what I do affects the rest of this > study. Most EM labs prefer that the user do a minimum of processing to the samples submitted.? We are a superstitious bunch and like to be able to control almost the entire embedding process.? That way if something gets messed up we can figure out what we might have done wrong and correct it. > > Thank you for your time and any valuable insights into this > matter. > > Chana de Wolf > Advanced Neural Biosciences, Inc. > From vapatpxs <@t> yahoo.com Wed May 6 11:36:25 2009 From: vapatpxs <@t> yahoo.com (Va Paula Sicurello) Date: Wed May 6 11:36:29 2009 Subject: [Histonet] Fixation for EM Message-ID: <562794.13458.qm@web46116.mail.sp1.yahoo.com> Oops,? My highlighted portions didn't make it through the filter.? The answers are after each question. :-} Paula Sicurello VA Medical Center San Diego Veterans Medical Research Foundation (VMRF) Core Research Imaging Center 3350 La Jolla Village Dr., MC151 San Diego, CA 92161 858-552-8585 x2397 From loftonjt <@t> holycrosshealth.org Wed May 6 13:08:20 2009 From: loftonjt <@t> holycrosshealth.org (Jimmy Lofton) Date: Wed May 6 13:08:37 2009 Subject: [Histonet] Liver Biopsy Concerns Message-ID: <4A0199D4.9B70.0056.0@holycrosshealth.org> Is any aware of the effects of anesthesia on the sample of liver biopsies? We have has some cutting and processing concerns regarding the use of certain anesthesia on our samples only coming on Ultra Sound specimens? Jimmy Lofton, M.S., HT,CT(ASCP) Manager Histology Laboratory Holy Cross Hospital 1500 Forest Glen Road Silver Spring, MD 20910-1484 301-754-7353 (Phone) 301-754-8563 (Fax) loftonjt@holycrosshealth.org Trinity Health MailGate made the following annotations --------------------------------------------------------------------- CONFIDENTIALITY NOTICE: This e-mail, including any attachments, is for the sole use of the intended recipient and may contain information that is confidential and privileged under state and Federal privacy laws. If you received this e-mail in error, be aware that any unauthorized use; disclosure, copying, or distribution is strictly prohibited. Please contact the sender immediately and destroy all copies of this message. --------------------------------------------------------------------- From laurie.colbert <@t> huntingtonhospital.com Wed May 6 13:31:35 2009 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Wed May 6 13:31:42 2009 Subject: [Histonet] CAP Question Message-ID: <57BE698966D5C54EAE8612E8941D76830580B88B@EXCHANGE3.huntingtonhospital.com> I would like to know how others interpret this CAP question ANP.21350: "Does the histology laboratory maintain records of the number of blocks, slides, and stains prepared" As far as the stains go - do you interpret this to mean do you keep track of the number of stains performed (as in, how many special stains do you do a year), or do you keep track of the number of stock stain solutions actually prepared? Laurie Colbert From gmartin <@t> marshallmedical.org Wed May 6 13:40:23 2009 From: gmartin <@t> marshallmedical.org (Martin, Gary) Date: Wed May 6 13:41:27 2009 Subject: [Histonet] CAP Question In-Reply-To: <57BE698966D5C54EAE8612E8941D76830580B88B@EXCHANGE3.huntingtonhospital.com> References: <57BE698966D5C54EAE8612E8941D76830580B88B@EXCHANGE3.huntingtonhospital.com> Message-ID: <6ED9D4252F278841A0593D3D788AF24C0537B73B@mailsvr.MARSHMED.local> Stains preformed -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert Sent: Wednesday, May 06, 2009 11:32 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CAP Question I would like to know how others interpret this CAP question ANP.21350: "Does the histology laboratory maintain records of the number of blocks, slides, and stains prepared" As far as the stains go - do you interpret this to mean do you keep track of the number of stains performed (as in, how many special stains do you do a year), or do you keep track of the number of stock stain solutions actually prepared? Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From billodonnell <@t> catholichealth.net Wed May 6 13:54:13 2009 From: billodonnell <@t> catholichealth.net (O'Donnell, Bill) Date: Wed May 6 13:54:31 2009 Subject: [Histonet] Charging for requested slides In-Reply-To: <6ED9D4252F278841A0593D3D788AF24C0537B73B@mailsvr.MARSHMED.local> References: <57BE698966D5C54EAE8612E8941D76830580B88B@EXCHANGE3.huntingtonhospital.com> <6ED9D4252F278841A0593D3D788AF24C0537B73B@mailsvr.MARSHMED.local> Message-ID: Greetings, >From time to time we get requests from law firms and other outside agencies to supply recuts and/or h&e slides. Sometimes these requests can be time-consuming. Is anyone out there charging for these services? William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 From jrobertson <@t> pathologysciences.com Wed May 6 14:13:27 2009 From: jrobertson <@t> pathologysciences.com (Jodie Robertson) Date: Wed May 6 14:13:38 2009 Subject: [Histonet] CAP Question In-Reply-To: <57BE698966D5C54EAE8612E8941D76830580B88B@EXCHANGE3.huntingtonhospital.com> References: <57BE698966D5C54EAE8612E8941D76830580B88B@EXCHANGE3.huntingtonhospital.com> Message-ID: <518CD6920AA7154193CBE5977CD880733A89AE@psmgsrv2.PSMG.local> How many slides you do a year is what we did and it passed CAP. We keep track of how many special stains, IHC and H&E a year in slides. Jodie Robertson, HT (ASCP) QIHC Pathology Sciences Medical Group -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert Sent: Wednesday, May 06, 2009 11:32 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CAP Question I would like to know how others interpret this CAP question ANP.21350: "Does the histology laboratory maintain records of the number of blocks, slides, and stains prepared" As far as the stains go - do you interpret this to mean do you keep track of the number of stains performed (as in, how many special stains do you do a year), or do you keep track of the number of stock stain solutions actually prepared? Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Wed May 6 14:14:02 2009 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Wed May 6 14:14:20 2009 Subject: [Histonet] Charging for requested slides In-Reply-To: References: <57BE698966D5C54EAE8612E8941D76830580B88B@EXCHANGE3.huntingtonhospital.com> <6ED9D4252F278841A0593D3D788AF24C0537B73B@mailsvr.MARSHMED.local> Message-ID: <4A01A93A.7400.0077.1@harthosp.org> Yes and, in addition, we collect payment before we do any of the work. Occasionally, after being informed of our charges, they cancel their request(s). Richard Richard W. Cartun, Ph.D. Director, Histology & Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "O'Donnell, Bill" 5/6/2009 2:54 PM >>> Greetings, >From time to time we get requests from law firms and other outside agencies to supply recuts and/or h&e slides. Sometimes these requests can be time-consuming. Is anyone out there charging for these services? William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jrobertson <@t> pathologysciences.com Wed May 6 14:14:53 2009 From: jrobertson <@t> pathologysciences.com (Jodie Robertson) Date: Wed May 6 14:15:03 2009 Subject: [Histonet] Charging for requested slides In-Reply-To: References: <57BE698966D5C54EAE8612E8941D76830580B88B@EXCHANGE3.huntingtonhospital.com><6ED9D4252F278841A0593D3D788AF24C0537B73B@mailsvr.MARSHMED.local> Message-ID: <518CD6920AA7154193CBE5977CD880733A89B0@psmgsrv2.PSMG.local> We charge for these. I believe we charge a nominal fee though. Jodie Robertson, HT (ASCP) QIHC Pathology Sciences Medical Group -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of O'Donnell, Bill Sent: Wednesday, May 06, 2009 11:54 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Charging for requested slides Greetings, >From time to time we get requests from law firms and other outside agencies to supply recuts and/or h&e slides. Sometimes these requests can be time-consuming. Is anyone out there charging for these services? William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GauchV <@t> mail.amc.edu Wed May 6 14:22:49 2009 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Wed May 6 14:23:29 2009 Subject: [Histonet] Charging for requested slides Message-ID: We charge for these requests also...we charge per slide and the stain they request determines the charge. Vicki Gauch,HTL (ASCP) Albany Medical Center Albany, NY >>> "O'Donnell, Bill" 5/6/2009 2:54 PM >>> Greetings, >From time to time we get requests from law firms and other outside agencies to supply recuts and/or h&e slides. Sometimes these requests can be time-consuming. Is anyone out there charging for these services? William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. From mucram11 <@t> comcast.net Wed May 6 14:27:55 2009 From: mucram11 <@t> comcast.net (Pamela Marcum) Date: Wed May 6 14:28:02 2009 Subject: [Histonet] Charging for requested slides In-Reply-To: <4A01A93A.7400.0077.1@harthosp.org> Message-ID: <1454328898.108691241638075437.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> I totally agree with getting the money upfront.? Think about it the lawyer is going to charge the client for getting the slides whether you do or not.? Your time is valuable and should be compensated by whatever hourly rate you feel you/your laboratory spent for the extra effort and reagents.? Don't give away your work and time. Pam Marcum UPENN Vet School New Bolton Center? ----- Original Message ----- From: "Richard Cartun" To: "Bill O'Donnell" , histonet@lists.utsouthwestern.edu Sent: Wednesday, May 6, 2009 3:14:02 PM GMT -05:00 US/Canada Eastern Subject: Re: [Histonet] Charging for requested slides Yes and, in addition, we collect payment before we do any of the work. ?Occasionally, after being informed of our charges, they cancel their request(s). Richard Richard W. Cartun, Ph.D. Director, Histology & Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT ?06102 (860) 545-1596 (860) 545-0174 Fax >>> "O'Donnell, Bill" 5/6/2009 2:54 PM >>> Greetings, >From time to time we get requests from law firms and other outside agencies to supply recuts and/or h&e slides. Sometimes these requests can be time-consuming. Is anyone out there charging for these services? William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 _______________________________________________ Histonet mailing list Histonet@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 tbritten <@t> aol.com Wed May 6 14:28:17 2009 From: tbritten <@t> aol.com (tbritten) Date: Wed May 6 14:33:42 2009 Subject: [Histonet] Charging for requested slides In-Reply-To: <1454328898.108691241638075437.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> References: <1454328898.108691241638075437.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> Message-ID: <55e0b058.a321.4a3a.9453.b7d09bdc89b0@aol.com> just an idea...has any one asked the lawyer what they charge their clients for these slides, etc...tom britten In a message dated 05/06/09 15:28:33 Eastern Daylight Time, mucram11@comcast.net writes: I totally agree with getting the money upfront. Think about it the lawyer is going to charge the client for getting the slides whether you do or not. Your time is valuable and should be compensated by whatever hourly rate you feel you/your laboratory spent for the extra effort and reagents. Don't give away your work and time. Pam Marcum UPENN Vet School New Bolton Center ----- Original Message ----- From: "Richard Cartun" To: "Bill O'Donnell" , histonet@lists.utsouthwestern.edu Sent: Wednesday, May 6, 2009 3:14:02 PM GMT -05:00 US/Canada Eastern Subject: Re: [Histonet] Charging for requested slides Yes and, in addition, we collect payment before we do any of the work. Occasionally, after being informed of our charges, they cancel their request(s). Richard Richard W. Cartun, Ph.D. Director, Histology & Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "O'Donnell, Bill" 5/6/2009 2:54 PM >>> Greetings, >From time to time we get requests from law firms and other outside agencies to supply recuts and/or h&e slides. Sometimes these requests can be time-consuming. Is anyone out there charging for these services? William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 _______________________________________________ Histonet mailing list Histonet@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 rjbuesa <@t> yahoo.com Wed May 6 15:05:02 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed May 6 15:05:08 2009 Subject: [Histonet] CAP Question In-Reply-To: <57BE698966D5C54EAE8612E8941D76830580B88B@EXCHANGE3.huntingtonhospital.com> Message-ID: <958871.85264.qm@web65710.mail.ac4.yahoo.com> That is a quantifying question by CAP and refers to keeping track of the number of blocks processed and slides stained either routine (H&E) or special procedures (HC, IHC)?daily to get to a monthly and annual figures. CAP does not care about the stocks prepared because how you use your stock solutions is another aspect of the QA. Ren? J. --- On Wed, 5/6/09, Laurie Colbert wrote: From: Laurie Colbert Subject: [Histonet] CAP Question To: histonet@lists.utsouthwestern.edu Date: Wednesday, May 6, 2009, 2:31 PM I would like to know how others interpret this CAP question ANP.21350: "Does the histology laboratory maintain records of the number of blocks, slides, and stains prepared" As far as the stains go - do you interpret this to mean do you keep track of the number of stains performed (as in, how many special stains do you do a year), or do you keep track of the number of stock stain solutions actually prepared? Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From billodonnell <@t> catholichealth.net Wed May 6 15:59:45 2009 From: billodonnell <@t> catholichealth.net (O'Donnell, Bill) Date: Wed May 6 15:59:57 2009 Subject: [Histonet] Charging for requested slides In-Reply-To: <55e0b058.a321.4a3a.9453.b7d09bdc89b0@aol.com> References: <1454328898.108691241638075437.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> <55e0b058.a321.4a3a.9453.b7d09bdc89b0@aol.com> Message-ID: Thank you al for your quick and informative responses. It seems to be universally accepted to charge for this service. You guys were a huge help. William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of tbritten Sent: Wednesday, May 06, 2009 2:28 PM To: Pamela Marcum; Richard Cartun Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Charging for requested slides just an idea...has any one asked the lawyer what they charge their clients for these slides, etc...tom britten In a message dated 05/06/09 15:28:33 Eastern Daylight Time, mucram11@comcast.net writes: I totally agree with getting the money upfront. Think about it the lawyer is going to charge the client for getting the slides whether you do or not. Your time is valuable and should be compensated by whatever hourly rate you feel you/your laboratory spent for the extra effort and reagents. Don't give away your work and time. Pam Marcum UPENN Vet School New Bolton Center ----- Original Message ----- From: "Richard Cartun" To: "Bill O'Donnell" , histonet@lists.utsouthwestern.edu Sent: Wednesday, May 6, 2009 3:14:02 PM GMT -05:00 US/Canada Eastern Subject: Re: [Histonet] Charging for requested slides Yes and, in addition, we collect payment before we do any of the work. Occasionally, after being informed of our charges, they cancel their request(s). Richard Richard W. Cartun, Ph.D. Director, Histology & Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "O'Donnell, Bill" 5/6/2009 2:54 PM >>> >>> Greetings, >From time to time we get requests from law firms and other outside agencies to supply recuts and/or h&e slides. Sometimes these requests can be time-consuming. Is anyone out there charging for these services? William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 _______________________________________________ Histonet mailing list Histonet@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 mwstarbu <@t> mdanderson.org Wed May 6 16:22:31 2009 From: mwstarbu <@t> mdanderson.org (Starbuck,Michael W) Date: Wed May 6 16:22:41 2009 Subject: [Histonet] TRAP staining and clast distribution Message-ID: <256363280A982F489DA7D9FDFD47DD8B170F4B26E7@DCPWVMBXC0VS3.mdanderson.edu> Hello, I'm doing a TRAP stain on MMA embedded undecalcified mouse tibia. I have noticed on a few groups of slides that all TRAP positive cells are localized on or near the cortex and growth plate and there are absolutely no positive cells in the mid-cancellous region of the proximal metaphasis. At the same time my controls work very well. Upon repeating the stain, the results come out the same. In my experience, I usually have TRAP positive cells show up throughout the cancellous bone. So, is the lack of TRAP positive cells in the cancellous bone of some slides a true distribution or could it possibly be due to inadequate penetration of fixative or a problem occurring during dehydration? Any help would be greatly appreciated. Thanks, Mike From mmcgraw <@t> phenopath.com Wed May 6 16:24:03 2009 From: mmcgraw <@t> phenopath.com (Medea McGraw) Date: Wed May 6 16:24:19 2009 Subject: [Histonet] wright-giemsa stain on tissue In-Reply-To: <1140541481.43fb4829095a0@webmail.tulane.edu> Message-ID: Renee, I was just looking in the histonet archives concerning doing a Wright Giemsa stain on FFPE tissue. We have a a FFPE cell pellet and was wondering the protocol on how you would stain this tissue? In the past, I have performed Wright Giemsa stains only on BAL?s / cytospins and therefore I am unsure of how to properly stain the researchers tissue. Any idea? Input on how to go about doing this? What if you were to rehydrate down to water and perform the test from there? Any suggestions would be greatly appreciated. Regards, Medea -- Medea J. McGraw BS, HT, HTL(ASCP) CM IHC Research Histotechnologist Phenopath Laboratories Phone: 206-374-9000 x1033 Fax: 206-374-9009 This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From SSCALISE <@t> beaumonthospitals.com Wed May 6 18:08:07 2009 From: SSCALISE <@t> beaumonthospitals.com (Sharon Scalise) Date: Wed May 6 18:24:00 2009 Subject: [Histonet] Soft LIS Message-ID: <4A01E017.1CA9.00C8.0@beaumonthospitals.com> Is anyone out there currently using Soft for their LIS? If so, are you utilizing slide and cassette labeling systems and/or specimen tracking at each step? I would be interested in speaking with anyone having experience with this system. Thank you! Sharon E. Scalise, HTL (ASCP) Histology Supervisor William Beaumont Hospital Royal Oak, MI 48073 248 898-5981 sscalise@beaumonthospitals.com From mburton1 <@t> bu.edu Wed May 6 18:35:36 2009 From: mburton1 <@t> bu.edu (mburton1@bu.edu) Date: Wed May 6 18:36:34 2009 Subject: [Histonet] Gram & AFB staining FNA smears In-Reply-To: <4A0156D6.2B7F.00C9.0@geisinger.edu> References: <4A0156D6.2B7F.00C9.0@geisinger.edu> Message-ID: <20090506193536.ez9kh85m3jswkc8c@www.bu.edu> Angela, The routine Gram stain procedure used in clinical micro labs is more simplified and is acceptable for staining thin smears but I don't believe it is suitable for tissue sections. So I would say the opposite is true of what the "doc" is saying: The stains used in the Histology lab would probably work fine for everything but I wouldn't send my tissue sections to the micro lab. Mark Burton HTL ASCP Mass General Hospital Boston, MA 02114 Quoting Angela Bitting : > The subject of staining FNA smears with AFB and Gram stains in > Histology vs in Micro came up today. Is there a reason that FNAs > can't be stained the same way we stain our tissue sections? One of > our docs was under the impression that it's not acceptable to use the > staining methods we use in our Histology lab. I don't know what > method Micro labs use, so I was hoping someone could shed some light > on this subject for me. > > Thanks, as always, > Angie > > Angela Bitting, HT(ASCP) > Technical Specialist, Histology > Geisinger Medical Center > 100 N Academy Ave. MC 23-00 > Danville, PA 17822 > phone 570-214-9634 > fax 570-271-5916 > > No trees were hurt in the sending of this email > However many electrons were severly inconvienienced! > > > > > IMPORTANT WARNING: The information in this message (and the documents > attached to it, if any) is confidential and may be legally > privileged. It is intended solely for the addressee. Access to this > message by anyone else is unauthorized. If you are not the intended > recipient, any disclosure, copying, distribution or any action taken, > or omitted to be taken, in reliance on it is prohibited and may be > unlawful. If you have received this message in error, please delete > all electronic copies of this message (and the documents attached to > it, if any), destroy any hard copies you may have created and notify > me immediately by replying to this email. Thank you. From Kemlo.Rogerson <@t> waht.swest.nhs.uk Thu May 7 02:07:09 2009 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu May 7 02:07:15 2009 Subject: [Histonet] Gram & AFB staining FNA smears Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06C0C5AD@wahtntex2.waht.swest.nhs.uk> Agree; I guess the difference is that FNAC and Micro samples haven't been subjected to processing like the tissue block sections. The tissue stains have been shown to work on something that has been fixed, dehydrated, set in hot wax, cut, rehydrated and then stained. The FNAC and Micro samples may or may not have been fixed (although I concede air drying is a form of fixation) and the stains used on them have been shown to work. The logic that these techniques are interchangeable is not only flawed but (oxy)moronic. Kemlo Rogerson e-mail kemlorogerson@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: 06 May 2009 17:24 To: histonet@lists.utsouthwestern.edu; Angela Bitting Subject: Re: [Histonet] Gram & AFB staining FNA smears The "doc" is wrong, otherwise your histology sections to be stained with Gram & AFB should also be sent to micro. Perhaps you will not have to do them anymore. Your "doc's" reasoning is purely oxymoronic. Ren? J. --- On Wed, 5/6/09, Angela Bitting wrote: From: Angela Bitting Subject: [Histonet] Gram & AFB staining FNA smears To: histonet@lists.utsouthwestern.edu Date: Wednesday, May 6, 2009, 9:22 AM The subject of staining FNA smears with AFB and Gram stains in Histology vs in Micro came up today. Is there a reason that FNAs can't be stained the same way we stain our tissue sections? One of our docs was under the impression that it's not acceptable to use the staining methods we use in our Histology lab. I don't know what method Micro labs use, so I was hoping someone could shed some light on this subject for me. Thanks, as always, Angie Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 No trees were hurt in the sending of this email However many electrons were severly inconvienienced! IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you._______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Thu May 7 07:53:33 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu May 7 07:53:39 2009 Subject: [Histonet] wright-giemsa stain on tissue In-Reply-To: Message-ID: <930170.81540.qm@web65704.mail.ac4.yahoo.com> Medea: Under separate cover I am sending an article I wrote on Giemsa for FFPE tissues. Ren? J. --- On Wed, 5/6/09, Medea McGraw wrote: From: Medea McGraw Subject: [Histonet] wright-giemsa stain on tissue To: histonet@lists.utsouthwestern.edu Date: Wednesday, May 6, 2009, 5:24 PM Renee, I was just looking in the histonet archives concerning doing a Wright Giemsa stain on FFPE tissue. We have a a FFPE cell pellet and was wondering the protocol on how you would stain this tissue? In the past, I have performed Wright Giemsa stains only on BAL?s / cytospins and therefore I am unsure of how to properly stain the researchers tissue. Any idea? Input on how to go about doing this? What if you were to rehydrate down to water and perform the test from there? Any suggestions would be greatly appreciated. Regards, Medea -- Medea J. McGraw BS, HT, HTL(ASCP) CM IHC Research Histotechnologist Phenopath Laboratories Phone: 206-374-9000 x1033 Fax: 206-374-9009 This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From arvidsonkristen <@t> yahoo.com Thu May 7 08:24:16 2009 From: arvidsonkristen <@t> yahoo.com (kristen arvidson) Date: Thu May 7 08:24:20 2009 Subject: [Histonet] Microtome Alignment Message-ID: <735163.62298.qm@web65716.mail.ac4.yahoo.com> Does anyone know where I can an alignment device? From jqb7 <@t> cdc.gov Thu May 7 08:30:25 2009 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine (CDC/CCID/NCZVED)) Date: Thu May 7 08:31:17 2009 Subject: [Histonet] Microtome Alignment In-Reply-To: <735163.62298.qm@web65716.mail.ac4.yahoo.com> References: <735163.62298.qm@web65716.mail.ac4.yahoo.com> Message-ID: <9A16CB5D55FC1648ADF11B63E72A1BE101AD3095@LTA3VS011.ees.hhs.gov> Newcomer sells them. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kristen arvidson Sent: Thursday, May 07, 2009 9:24 AM To: histonet Subject: [Histonet] Microtome Alignment Does anyone know where I can an alignment device? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akemiat3377 <@t> yahoo.com Thu May 7 08:40:25 2009 From: akemiat3377 <@t> yahoo.com (Akemi Allison-Tacha) Date: Thu May 7 08:40:29 2009 Subject: [Histonet] Microtome Alignment Message-ID: <376431.39678.qm@web31303.mail.mud.yahoo.com> David Davis from CO designed an alignment device and can be purchased from Newcomer supply or I think his e-mail is still the following:?daviddavis@uchsc.edu Good Luck, Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Ct. Los Gatos, CA 95032 Cell: 425.941.4287 W: E-Mail: aallison-tacha@apmglab.com P: E-Mail: akemiat3377@yahoo.com --- On Thu, 5/7/09, kristen arvidson wrote: From: kristen arvidson Subject: [Histonet] Microtome Alignment To: "histonet" Date: Thursday, May 7, 2009, 6:24 AM Does anyone know where I can an alignment device? ? ? ? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lmnichols <@t> shaw.ca Fri May 1 14:42:43 2009 From: lmnichols <@t> shaw.ca (Leah Nichols) Date: Thu May 7 08:49:35 2009 Subject: [Histonet] What are peoples feelings on pre labling slides? Message-ID: <49FB50B3.3000603@shaw.ca> From lmnichols <@t> shaw.ca Fri May 1 15:06:08 2009 From: lmnichols <@t> shaw.ca (Leah Nichols) Date: Thu May 7 08:49:37 2009 Subject: [Histonet] What are peoples feelings on pre labling slides? Message-ID: <49FB5630.7040904@shaw.ca> An independent QA study was completed in our AP department. One of the report's recommendations was that we do not pre-label our slides and develop a "just in time" slide labelling process. We are now looking into this and would appreciate your feedback/comments on "just in time" slide labelling. The questions I have for labs that has developed their own "just in time" slide labelling process are: * Who labels your slides? * When do you label your slides? * What is your process? * Has this decreased your error rate? * Off topic - when do you label your cassettes? Thanks, Leah Nichols Lean Facilitator Calgary Laboratory Services From kfeaster <@t> hsc.wvu.edu Mon May 4 08:55:35 2009 From: kfeaster <@t> hsc.wvu.edu (Kimberly Feaster) Date: Thu May 7 08:49:39 2009 Subject: [Histonet] New HTL Program! Message-ID: <49FEBB97.6676.0078.0@hsc.wvu.edu> West Virginia University School of Medicine Histotechnology Program is now accepting applications for fall 2009 enrollment! The WVU Histotechnology Program is offered as an area of emphasis within the Medical Laboratory Science major. Students graduate with a Bachelor?s of Science degree in Medical Laboratory Science. The first two years consist of a pre-professional curriculum and the last two years consist of the Histotechnology Program curriculum. The pre-professional curriculum can be completed at WVU main campus, one of the WVU regional campuses or any regionally accredited college or university. The Histotechnology Program curriculum is based at the WVU Robert C. Byrd Health Sciences Center. The first year consists of a didactic schedule focusing on routine laboratory procedures with incorporated laboratory sessions and the second year will focus on complex procedures with on-site and clinical rotations. Clinical rotations will be completed at the program?s affiliated clinical laboratories located in West Virginia and Pennsylvania. If anyone would like more information or know someone who does, please contact: Kimberly Feaster Histotechnology Program Director 304-293-7628 kfeaster@hsc.wvu.edu From jfinke <@t> unmc.edu Mon May 4 15:48:09 2009 From: jfinke <@t> unmc.edu (Jennifer S Finke) Date: Thu May 7 08:49:41 2009 Subject: [Histonet] Cryo question Message-ID: To whom it may concern: I have a question about OTC emb sometime ago and kept at -20C. Is there a and re-hydrate the tissue? Thank you, Jennifer Finke-Dwyer Department of Pharmacology and Experi University of Nebraska Medical Center 985880 Nebr Omaha, NE 68198-5880 Phone: (402)559-8925 From LSebree <@t> uwhealth.org Thu May 7 08:57:03 2009 From: LSebree <@t> uwhealth.org (Sebree Linda A) Date: Thu May 7 08:57:10 2009 Subject: [Histonet] New HTL Program! In-Reply-To: <49FEBB97.6676.0078.0@hsc.wvu.edu> Message-ID: <5998F3BDFF7AAC4091C7AE93A7A1A5892CDFC5@UWHC-MAIL01.uwhis.hosp.wisc.edu> This is great news for our field! If I were 30 years younger..... Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kimberly Feaster Sent: Monday, May 04, 2009 8:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New HTL Program! West Virginia University School of Medicine Histotechnology Program is now accepting applications for fall 2009 enrollment! The WVU Histotechnology Program is offered as an area of emphasis within the Medical Laboratory Science major. Students graduate with a Bachelor's of Science degree in Medical Laboratory Science. The first two years consist of a pre-professional curriculum and the last two years consist of the Histotechnology Program curriculum. The pre-professional curriculum can be completed at WVU main campus, one of the WVU regional campuses or any regionally accredited college or university. The Histotechnology Program curriculum is based at the WVU Robert C. Byrd Health Sciences Center. The first year consists of a didactic schedule focusing on routine laboratory procedures with incorporated laboratory sessions and the second year will focus on complex procedures with on-site and clinical rotations. Clinical rotations will be completed at the program's affiliated clinical laboratories located in West Virginia and Pennsylvania. If anyone would like more information or know someone who does, please contact: Kimberly Feaster Histotechnology Program Director 304-293-7628 kfeaster@hsc.wvu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Thu May 7 09:02:49 2009 From: mucram11 <@t> comcast.net (Pamela Marcum) Date: Thu May 7 09:02:53 2009 Subject: [Histonet] New HTL Program! In-Reply-To: <5998F3BDFF7AAC4091C7AE93A7A1A5892CDFC5@UWHC-MAIL01.uwhis.hosp.wisc.edu> Message-ID: <1227696573.35171241704969722.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> This is great news!!? Good luck and let's all wish this school success for many in years into the future.? It sounds like a very good program. Pam Marcum UPENN Vet School New Bolton Center ----- Original Message ----- From: "Sebree Linda A" To: "Kimberly Feaster" , histonet@lists.utsouthwestern.edu Sent: Thursday, May 7, 2009 9:57:03 AM GMT -05:00 US/Canada Eastern Subject: RE: [Histonet] New HTL Program! This is great news for our field! ?If I were 30 years younger..... Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kimberly Feaster Sent: Monday, May 04, 2009 8:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New HTL Program! West Virginia University School of Medicine Histotechnology Program is now accepting applications for fall 2009 enrollment! The WVU Histotechnology Program is offered as an area of emphasis within the Medical Laboratory Science major. ?Students graduate with a Bachelor's of Science degree in Medical Laboratory Science. ? The first two years consist of a pre-professional curriculum and the last two years consist of the Histotechnology Program curriculum. ? The pre-professional curriculum can be completed at WVU main campus, one of the WVU regional campuses or any regionally accredited college or university. ? The Histotechnology Program curriculum is based at the WVU Robert C. Byrd Health Sciences Center. ?The first year consists of a didactic schedule focusing on routine laboratory procedures with incorporated laboratory sessions and the second year will focus on complex procedures with on-site and clinical rotations. ?Clinical rotations will be completed at the program's affiliated clinical laboratories located in West Virginia and Pennsylvania. ? If anyone would like more information or know someone who does, please contact: Kimberly Feaster Histotechnology Program Director 304-293-7628 kfeaster@hsc.wvu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Shakun.Aswani <@t> acologix.com Thu May 7 09:08:54 2009 From: Shakun.Aswani <@t> acologix.com (Shakun Aswani) Date: Thu May 7 09:08:56 2009 Subject: [Histonet] New HTL Program! In-Reply-To: <49FEBB97.6676.0078.0@hsc.wvu.edu> References: <49FEBB97.6676.0078.0@hsc.wvu.edu> Message-ID: <777AB0DE519C8E46A6220E2287C5BAD301B3535F@EXCHANGE.acologix.com> Fantastic!!! I was waiting for this happen. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kimberly Feaster Sent: Monday, May 04, 2009 6:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New HTL Program! West Virginia University School of Medicine Histotechnology Program is now accepting applications for fall 2009 enrollment! The WVU Histotechnology Program is offered as an area of emphasis within the Medical Laboratory Science major. Students graduate with a Bachelor's of Science degree in Medical Laboratory Science. The first two years consist of a pre-professional curriculum and the last two years consist of the Histotechnology Program curriculum. The pre-professional curriculum can be completed at WVU main campus, one of the WVU regional campuses or any regionally accredited college or university. The Histotechnology Program curriculum is based at the WVU Robert C. Byrd Health Sciences Center. The first year consists of a didactic schedule focusing on routine laboratory procedures with incorporated laboratory sessions and the second year will focus on complex procedures with on-site and clinical rotations. Clinical rotations will be completed at the program's affiliated clinical laboratories located in West Virginia and Pennsylvania. If anyone would like more information or know someone who does, please contact: Kimberly Feaster Histotechnology Program Director 304-293-7628 kfeaster@hsc.wvu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ian.montgomery <@t> bio.gla.ac.uk Thu May 7 09:11:01 2009 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Thu May 7 09:13:51 2009 Subject: FW: [Histonet] Microtome Alignment Message-ID: <69829D0E45844D9188B51C22728DEA08@IBLS.GLA.AC.UK> What on the microtome do you want to align? Dr. Ian Montgomery, Histotechnology, I.B.L.S. Support Unit, Thomson Building, University of Glasgow, Glasgow, G12 8QQ. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kristen arvidson Sent: 07 May 2009 14:24 To: histonet Subject: [Histonet] Microtome Alignment Does anyone know where I can an alignment device? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> waht.swest.nhs.uk Thu May 7 09:17:34 2009 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu May 7 09:17:41 2009 Subject: [Histonet] New HTL Program! Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06C0C68B@wahtntex2.waht.swest.nhs.uk> Congratulations from the UK. As you know we've had this in place for many years and resulted in the profession having a higher profile; I hope the rest will follow for you. Kemlo Rogerson e-mail kemlorogerson@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pamela Marcum Sent: 07 May 2009 15:03 To: Sebree Linda A Cc: histonet@lists.utsouthwestern.edu; Kimberly Feaster Subject: Re: [Histonet] New HTL Program! This is great news!!? Good luck and let's all wish this school success for many in years into the future.? It sounds like a very good program. Pam Marcum UPENN Vet School New Bolton Center ----- Original Message ----- From: "Sebree Linda A" To: "Kimberly Feaster" , histonet@lists.utsouthwestern.edu Sent: Thursday, May 7, 2009 9:57:03 AM GMT -05:00 US/Canada Eastern Subject: RE: [Histonet] New HTL Program! This is great news for our field! ?If I were 30 years younger..... Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kimberly Feaster Sent: Monday, May 04, 2009 8:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New HTL Program! West Virginia University School of Medicine Histotechnology Program is now accepting applications for fall 2009 enrollment! The WVU Histotechnology Program is offered as an area of emphasis within the Medical Laboratory Science major. ?Students graduate with a Bachelor's of Science degree in Medical Laboratory Science. ? The first two years consist of a pre-professional curriculum and the last two years consist of the Histotechnology Program curriculum. ? The pre-professional curriculum can be completed at WVU main campus, one of the WVU regional campuses or any regionally accredited college or university. The Histotechnology Program curriculum is based at the WVU Robert C. Byrd Health Sciences Center. ?The first year consists of a didactic schedule focusing on routine laboratory procedures with incorporated laboratory sessions and the second year will focus on complex procedures with on-site and clinical rotations. ?Clinical rotations will be completed at the program's affiliated clinical laboratories located in West Virginia and Pennsylvania. ? If anyone would like more information or know someone who does, please contact: Kimberly Feaster Histotechnology Program Director 304-293-7628 kfeaster@hsc.wvu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From smcbride <@t> cs.cmu.edu Thu May 7 10:04:54 2009 From: smcbride <@t> cs.cmu.edu (Sean McBride) Date: Thu May 7 10:05:02 2009 Subject: [Histonet] Request for Staining Advice: Rabbit Calvaria & underlying Cerebral Hemishpere Message-ID: Colleagues, I have been charged with the task of evaluating an hydroxyapatite coated biomaterial implanted in the calvaria of a rabbit model. The calvaria and cerebral hemisphere were excised en bloc, and the objective of the experiment is to assess not only new bone formation and osseointegration but also any pathological effects on the dura mater, arachnoid, pia mater and underlying cerebral hemisphere. I have assessed both brain and bone individually in the past, but never en bloc. The specimens have been embedded in pmma and mounted for thick ground section (~30 ? 35 microns) evaluation. Does anyone have any suggestions for staining techniques that will show both bone & brain morphology in an en bloc specimen? Kind regards, ~Sean McBride Senior Researcher Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-901-7540 (m) 412-268-8641 (fax) smcbride@cs.cmu.edu From Shakun.Aswani <@t> acologix.com Thu May 7 10:13:15 2009 From: Shakun.Aswani <@t> acologix.com (Shakun Aswani) Date: Thu May 7 10:13:19 2009 Subject: [Histonet] Request for Staining Advice: Rabbit Calvaria & underlying Cerebral Hemishpere In-Reply-To: References: Message-ID: <777AB0DE519C8E46A6220E2287C5BAD301B35360@EXCHANGE.acologix.com> Try Goldner's Trichrome Stain. I have protocol if you want -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sean McBride Sent: Thursday, May 07, 2009 8:05 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Request for Staining Advice: Rabbit Calvaria & underlying Cerebral Hemishpere Colleagues, I have been charged with the task of evaluating an hydroxyapatite coated biomaterial implanted in the calvaria of a rabbit model. The calvaria and cerebral hemisphere were excised en bloc, and the objective of the experiment is to assess not only new bone formation and osseointegration but also any pathological effects on the dura mater, arachnoid, pia mater and underlying cerebral hemisphere. I have assessed both brain and bone individually in the past, but never en bloc. The specimens have been embedded in pmma and mounted for thick ground section (~30 - 35 microns) evaluation. Does anyone have any suggestions for staining techniques that will show both bone & brain morphology in an en bloc specimen? Kind regards, ~Sean McBride Senior Researcher Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-901-7540 (m) 412-268-8641 (fax) smcbride@cs.cmu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From b-frederick <@t> northwestern.edu Thu May 7 10:16:31 2009 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Thu May 7 10:16:46 2009 Subject: [Histonet] Microtome Alignment In-Reply-To: <735163.62298.qm@web65716.mail.ac4.yahoo.com> Message-ID: Microm sells a device called a histocollimeter, however, it only attaches to their microtomes to my knowledge. Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility ECOGPCO-RL 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kristen arvidson Sent: Thursday, May 07, 2009 8:24 AM To: histonet Subject: [Histonet] Microtome Alignment Does anyone know where I can an alignment device? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NMargaryan <@t> childrensmemorial.org Thu May 7 10:23:08 2009 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Thu May 7 10:24:30 2009 Subject: [Histonet] FW: prover' pojaluista Message-ID: Dear Histonetters, This is my question regarding IHC staining and water temperature: does IHC staining depend on water temperature in washing step? As always, I am using the same protocol for IHC. But one day it works perfectly, another day, on the same tissue, it does not work. Everything is same, solutions are fresh, and the only thing that is different is water temperature. I am using running distilled water before and after Antigen Retrieval step, after DAB, after counterstaining and after bluing reagent. Dd-water goes into autostainer: all blocking steps and after primary, secondary and tertiary. I am not using usual running water to exclude different Chlorine concentrations in different days. Have anybody of you ever paid attention to running distilled water temperature? What water temperature (cold or warm) you will recommend for washing steps? If this is critical, I can prepare my water up front. Hope to get an answer soon. Thanks, Naira From jrobertson <@t> pathologysciences.com Thu May 7 11:03:36 2009 From: jrobertson <@t> pathologysciences.com (Jodie Robertson) Date: Thu May 7 11:03:40 2009 Subject: [Histonet] Microtome Alignment In-Reply-To: <735163.62298.qm@web65716.mail.ac4.yahoo.com> References: <735163.62298.qm@web65716.mail.ac4.yahoo.com> Message-ID: <518CD6920AA7154193CBE5977CD880733A89B8@psmgsrv2.PSMG.local> Newcomer Supply. Jodie Robertson, HT (ASCP) QIHC Pathology Sciences Medical Group -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kristen arvidson Sent: Thursday, May 07, 2009 6:24 AM To: histonet Subject: [Histonet] Microtome Alignment Does anyone know where I can an alignment device? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From koswalt <@t> deltapathology.com Thu May 7 11:10:13 2009 From: koswalt <@t> deltapathology.com (koswalt@deltapathology.com) Date: Thu May 7 11:10:17 2009 Subject: [Histonet] how to unregister from histo-net Message-ID: <20090507111013.wkmxja2foc4gcggc@mail3.deltapathology.com> Could anyone help me. I am trying to un-listg with the histo-net services. Please let me know. Thanks, Kim From koswalt <@t> deltapathology.com Thu May 7 11:15:41 2009 From: koswalt <@t> deltapathology.com (koswalt@deltapathology.com) Date: Thu May 7 11:15:44 2009 Subject: [Histonet] Remove from e-mail Message-ID: <20090507111541.eu7fgeov4k404w84@mail3.deltapathology.com> Please remove me from your e-mail lists. Thank You, Kim From victor <@t> pathology.washington.edu Thu May 7 11:21:23 2009 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Thu May 7 11:21:28 2009 Subject: [Histonet] Microtome Alignment In-Reply-To: <518CD6920AA7154193CBE5977CD880733A89B8@psmgsrv2.PSMG.local> References: <735163.62298.qm@web65716.mail.ac4.yahoo.com> <518CD6920AA7154193CBE5977CD880733A89B8@psmgsrv2.PSMG.local> Message-ID: <4A030A83.30802@pathology.washington.edu> I just went to Newcomer Supply to see what you get for $700, not much. Go to Ace Hardware and get each employee one of these. It will work just as well and works on any microtome. Buy your staff lunch with what you save. http://www.acehardware.com/product/index.jsp?productId=1286367&cp=&fbc=1&pg=3&kw=level&lmdn=Price+Range&fbn=StorePrice%7CUnder+%2425.00&fr=StorePrice%2FACE%2F00000000%2F00002500&parentPage=search&searchId=37640061873 Victor Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. Jodie Robertson wrote: > Newcomer Supply. > > > Jodie Robertson, HT (ASCP) QIHC > Pathology Sciences Medical Group > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kristen > arvidson > Sent: Thursday, May 07, 2009 6:24 AM > To: histonet > Subject: [Histonet] Microtome Alignment > > Does anyone know where I can an alignment device? > > > > _______________________________________________ > Histonet mailing list > Histonet@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 ebreisch <@t> rchsd.org Thu May 7 11:23:39 2009 From: ebreisch <@t> rchsd.org (Breisch, Eric) Date: Thu May 7 11:23:45 2009 Subject: [Histonet] NADPH-diaphorase Message-ID: <43B97B4C402C2C44AAA2A8D2C86A88B303A52F72@e2k3backend1.RCHSD.org> Hi, hoping that someone might be able to share with us where they purchase NADPH-diaphorase for the NADPH stain. Thank you. EAB Eric A. Breisch, Ph.D. Clinical Anatomist Dept. of Pathology Rady Children's Hospital and Health Center Associate Clinical Professor of Anatomy Dept. of Surgery UCSD School of Medicine From portera <@t> msu.edu Thu May 7 11:32:00 2009 From: portera <@t> msu.edu (Amy Porter) Date: Thu May 7 11:32:07 2009 Subject: [Histonet] NADPH-diaphorase References: <43B97B4C402C2C44AAA2A8D2C86A88B303A52F72@e2k3backend1.RCHSD.org> Message-ID: <017BD9C91FC84ECEAF5D480A402ABA54@histolab> Try Sigma-Aldrich. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: portera@msu.edu Web: www.humanpathology.msu.edu ----- Original Message ----- From: "Breisch, Eric" To: Sent: Thursday, May 07, 2009 12:23 PM Subject: [Histonet] NADPH-diaphorase Hi, hoping that someone might be able to share with us where they purchase NADPH-diaphorase for the NADPH stain. Thank you. EAB Eric A. Breisch, Ph.D. Clinical Anatomist Dept. of Pathology Rady Children's Hospital and Health Center Associate Clinical Professor of Anatomy Dept. of Surgery UCSD School of Medicine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gmartin <@t> marshallmedical.org Thu May 7 11:37:52 2009 From: gmartin <@t> marshallmedical.org (Martin, Gary) Date: Thu May 7 11:38:53 2009 Subject: [Histonet] Microtome Alignment In-Reply-To: <4A030A83.30802@pathology.washington.edu> References: <735163.62298.qm@web65716.mail.ac4.yahoo.com><518CD6920AA7154193CBE5977CD880733A89B8@psmgsrv2.PSMG.local> <4A030A83.30802@pathology.washington.edu> Message-ID: <6ED9D4252F278841A0593D3D788AF24C0537C1F8@mailsvr.MARSHMED.local> As an old carpenter I quickly adapted a "speed square" to do pretty much the same thing as the $775.00 unit or the torpedo level. The speed square has the advantage of squaring the block holder to a true flat surface (the base of the microtome) it works quite well ... it is positive and quick. See the Ace website below. I also use the torpedo level as a double check. http://www.acehardware.com/search/index.jsp?kwCatId=&kw=speed%20square&o rigkw=speed%20square&sr=1 Gary (in California) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Victor Tobias Sent: Thursday, May 07, 2009 9:21 AM To: Jodie Robertson Cc: histonet Subject: Re: [Histonet] Microtome Alignment I just went to Newcomer Supply to see what you get for $700, not much. Go to Ace Hardware and get each employee one of these. It will work just as well and works on any microtome. Buy your staff lunch with what you save. http://www.acehardware.com/product/index.jsp?productId=1286367&cp=&fbc=1 &pg=3&kw=level&lmdn=Price+Range&fbn=StorePrice%7CUnder+%2425.00&fr=Store Price%2FACE%2F00000000%2F00002500&parentPage=search&searchId=37640061873 Victor Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. Jodie Robertson wrote: > Newcomer Supply. > > > Jodie Robertson, HT (ASCP) QIHC > Pathology Sciences Medical Group > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kristen > arvidson > Sent: Thursday, May 07, 2009 6:24 AM > To: histonet > Subject: [Histonet] Microtome Alignment > > Does anyone know where I can an alignment device? > > > > _______________________________________________ > Histonet mailing list > Histonet@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 rjbuesa <@t> yahoo.com Thu May 7 11:57:18 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu May 7 11:57:22 2009 Subject: [Histonet] FW: prover' pojaluista In-Reply-To: Message-ID: <766484.83301.qm@web65704.mail.ac4.yahoo.com> Strictly speaking, IHC reactions do not depend on water temperature temperature. What you describe does not include dist. water between the IHC steps therefore the water temp. should not be affecting your reactions. If you use dist water after HIER you have to remember placing the slides?into room temp. buffer before starting the IHC procedure, so if there was any effect due to the dist. water, that should have take care of it. The inconsistency in your staining I think is not due to the dist. water temp. Ren? J. --- On Thu, 5/7/09, Margaryan, Naira wrote: From: Margaryan, Naira Subject: [Histonet] FW: prover' pojaluista To: "histonet@lists.utsouthwestern.edu" Date: Thursday, May 7, 2009, 11:23 AM Dear Histonetters, This is my question regarding IHC staining and water temperature: does IHC staining depend on water temperature in washing step? As always, I am using the same protocol for IHC. But one day it works perfectly, another day, on the same tissue, it does not work. Everything is same, solutions are fresh, and the only thing that is different is water temperature. I am using running distilled water before and after Antigen Retrieval step, after DAB, after counterstaining and after bluing reagent. Dd-water goes into autostainer: all blocking steps and after primary, secondary and tertiary. I am not using usual running water to exclude different Chlorine concentrations in different days. Have anybody of you ever paid attention to running distilled water temperature? What water temperature (cold or warm) you will recommend for washing steps? If this is critical, I can prepare my water up front. Hope to get an answer soon. Thanks, Naira _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Timothy.Morken <@t> ucsfmedctr.org Thu May 7 12:10:12 2009 From: Timothy.Morken <@t> ucsfmedctr.org (Morken, Tim) Date: Thu May 7 12:10:25 2009 Subject: Make a Tiny URL of that thing...RE: [Histonet] Microtome Alignment In-Reply-To: <4A030A83.30802@pathology.washington.edu> References: <735163.62298.qm@web65716.mail.ac4.yahoo.com> <518CD6920AA7154193CBE5977CD880733A89B8@psmgsrv2.PSMG.local> <4A030A83.30802@pathology.washington.edu> Message-ID: <1AAF670737F193429070841C6B2ADD4C01C2F7B5@EXMBMCB15.ucsfmedicalcenter.org> Victor wrote: "Go to Ace Hardware and get each employee one of these... http://www.acehardware.com/product/index.jsp?productId=1286367&cp=&fbc=1&pg=3&kw=level&lmdn=Price+Range&fbn=StorePrice%7CUnder+%2425.00&fr=StorePrice%2FACE%2F00000000%2F00002500&parentPage=search&searchId=37640061873 " You all need to learn about TinyURL.com. You can paste in a large URL and it makes tiny one that is easy to use in emails. Sometimes the long ones get cut up in email and don't work Here is the same link as above, but after using tinyURL.com: http://tinyurl.com/d9y3s9 Tim Morken Supervisor, Histology / IPOX UCSF Medical Center San Francisco, CA From gmartin <@t> marshallmedical.org Thu May 7 12:17:58 2009 From: gmartin <@t> marshallmedical.org (Martin, Gary) Date: Thu May 7 12:18:54 2009 Subject: Make a Tiny URL of that thing...RE: [Histonet] Microtome Alignment In-Reply-To: <1AAF670737F193429070841C6B2ADD4C01C2F7B5@EXMBMCB15.ucsfmedicalcenter.org> References: <735163.62298.qm@web65716.mail.ac4.yahoo.com><518CD6920AA7154193CBE5977CD880733A89B8@psmgsrv2.PSMG.local><4A030A83.30802@pathology.washington.edu> <1AAF670737F193429070841C6B2ADD4C01C2F7B5@EXMBMCB15.ucsfmedicalcenter.org> Message-ID: <6ED9D4252F278841A0593D3D788AF24C0537C2F8@mailsvr.MARSHMED.local> Thank you Tim ... very cool! Gary -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim Sent: Thursday, May 07, 2009 10:10 AM To: histonet Subject: Make a Tiny URL of that thing...RE: [Histonet] Microtome Alignment Victor wrote: "Go to Ace Hardware and get each employee one of these... http://www.acehardware.com/product/index.jsp?productId=1286367&cp=&fbc=1 &pg=3&kw=level&lmdn=Price+Range&fbn=StorePrice%7CUnder+%2425.00&fr=Store Price%2FACE%2F00000000%2F00002500&parentPage=search&searchId=37640061873 " You all need to learn about TinyURL.com. You can paste in a large URL and it makes tiny one that is easy to use in emails. Sometimes the long ones get cut up in email and don't work Here is the same link as above, but after using tinyURL.com: http://tinyurl.com/d9y3s9 Tim Morken Supervisor, Histology / IPOX UCSF Medical Center San Francisco, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Nikki.Wahlberg <@t> bsci.com Thu May 7 12:38:02 2009 From: Nikki.Wahlberg <@t> bsci.com (Wahlberg, Nikki) Date: Thu May 7 12:38:10 2009 Subject: [Histonet] Turnbull's Blue Message-ID: <5BF2F9FB24C0DC499CD3C4BB6F7B4481013E23FF@MAPMAIL02.bsci.bossci.com> Is anyone performing the Turnbull's Blue Iron stain? If so, what type of control do you use. We are having a difficult time finding a good control in order to get the stain approved in our lab. Also, if anyone has some tips for the procedure they would be greatly appreciated. Thank you Nikki M Wahlberg Histotechnologist II Histo-Pathology Services Boston Scientific 763-694-5739 wahlbern@bsci.com CONFIDENTIALITY NOTICE: This e-mail transmission may contain confidential or legally privileged information that is intended only for the individual or entity named in the e-mail address. Any unauthorized distribution or copying of this transmittal or its attachments, if any, is prohibited. If you have received this e-mail transmission in error, please reply to the sender, so that Boston Scientific Corporation can arrange for proper delivery, then please delete the message from your inbox. From Bryan.Watson <@t> parkview.com Thu May 7 12:49:49 2009 From: Bryan.Watson <@t> parkview.com (Bryan Watson) Date: Thu May 7 12:50:18 2009 Subject: [Histonet] pinning specimens for fixation Message-ID: <4A02E6FC.5674.0085.1@parkview.com> We have a surgeon who likes to send us specimens that are pinned down to a board of some sort. He uses whatever makeshift board he can find and he wants us to order something for that purpose. Does anybody use anything like this something strong enough to hold a specimen pinned down without the pins being pulled out that is also disposable? If so, can you tell me what they're called and where to get them, thanks. I'll take any alternate ideas as well. Thanks, Bryan From secash <@t> gundluth.org Thu May 7 12:56:16 2009 From: secash <@t> gundluth.org (secash@gundluth.org) Date: Thu May 7 12:56:22 2009 Subject: [Histonet] Holding Cell Lines in Methanol Message-ID: We are doing a 5 day Hypoxia Chamber Experiment on Cancer Cell Lines Slides. They are fixed in methanol at -20c for 20 minutes when they come out. If I cannot do the experiment at that time (due to a funeral), can these slides be stored in the methanol at -20c overnight without it adversely harming the cells? Thanks for the help!! Steven Cash, BS, HT Supervisor Oncology Research Lab Gundersen Lutheran Medical Foundation 1300 Badger St LaCrosse, WI 54601 608-775-3548 Mail Stop UWL-002 From juditw <@t> u.washington.edu Thu May 7 13:05:00 2009 From: juditw <@t> u.washington.edu (Judith L. Williams) Date: Thu May 7 13:05:13 2009 Subject: [Histonet] pinning specimens for fixation In-Reply-To: <4A02E6FC.5674.0085.1@parkview.com> Message-ID: HI Bryan - In biology laboratories they use metal trays filled with a wax like product. You can make your own with a 9x12 metal baking dish filled half full with paraffin. Or, you can order the "professional" ones from Carolina Biological Supply, cheaper than from the anatomic path catalogs. They also have the large T-shape pins. good luck with the dissection and fixation! Judy On Thu, 7 May 2009, Bryan Watson wrote: > We have a surgeon who likes to send us specimens that are pinned down to a board of some sort. He uses whatever makeshift board he can find and he wants us to order something for that purpose. Does anybody use anything like this something strong enough to hold a specimen pinned down without the pins being pulled out that is also disposable? If so, can you tell me what they're called and where to get them, thanks. I'll take any alternate ideas as well. > Thanks, > Bryan > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Judith Williams, PhD, HT(ASCP) Research Scientist Department of Comparative Medicine University of Washington Seattle, WA 98195 From lori.garcia <@t> medtronic.com Thu May 7 13:10:20 2009 From: lori.garcia <@t> medtronic.com (Garcia, Lori, Sr. Scientist) Date: Thu May 7 13:10:26 2009 Subject: [Histonet] pinning specimens for fixation In-Reply-To: <4A02E6FC.5674.0085.1@parkview.com> References: <4A02E6FC.5674.0085.1@parkview.com> Message-ID: <0F844FC5FD7212428416B8565B47FA0202C85691@STSM1BMSGM03.ent.core.medtronic.com> Hi Bryan, Mopec makes disposable cutting boards that can be cut to the right size. Here is the link: http://www.mopec.com/product/626/small_disposable_cutting_boards/ Also, thick cork board works very well. Lori -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Watson Sent: Thursday, May 07, 2009 10:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pinning specimens for fixation We have a surgeon who likes to send us specimens that are pinned down to a board of some sort. He uses whatever makeshift board he can find and he wants us to order something for that purpose. Does anybody use anything like this something strong enough to hold a specimen pinned down without the pins being pulled out that is also disposable? If so, can you tell me what they're called and where to get them, thanks. I'll take any alternate ideas as well. Thanks, Bryan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet [CONFIDENTIALITY AND PRIVACY NOTICE] Information transmitted by this email is proprietary to Medtronic and is intended for use only by the individual or entity to which it is addressed, and may contain information that is private, privileged, confidential or exempt from disclosure under applicable law. If you are not the intended recipient or it appears that this mail has been forwarded to you without proper authority, you are notified that any use or dissemination of this information in any manner is strictly prohibited. In such cases, please delete this mail from your records. To view this notice in other languages you can either select the following link or manually copy and paste the link into the address bar of a web browser: http://emaildisclaimer.medtronic.com From Maxim_71 <@t> mail.ru Thu May 7 13:41:23 2009 From: Maxim_71 <@t> mail.ru (Maxim Peshkov) Date: Thu May 7 13:42:11 2009 Subject: [Histonet] Gram & AFB staining FNA smears Message-ID: <1082162638.20090507224123@mail.ru> Angela: We stains our cytology slides for Gram and AFB with the same reagents as histology. Only misses dewax and final dehydration, clearing and coverslip steps. Results are like both, the pathologists and cytopathologist. Maxim Peshkov, Russia, Taganrog. mailto:Maxim_71@mail.ru From llewllew <@t> shaw.ca Thu May 7 13:47:53 2009 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Thu May 7 13:49:02 2009 Subject: [Histonet] pinning specimens for fixation In-Reply-To: <4A02E6FC.5674.0085.1@parkview.com> References: <4A02E6FC.5674.0085.1@parkview.com> Message-ID: <0AB71D82DDEC4FFC95EB895BAB33A210@BryanPC> Try coroplast panels from your local home renovation/hardware store and cut to the size you want with a utility knife. Also, make sure that the surgeon uses plastic pins, not steel, to avoid rust. Since they will probably float, turn them over so the tissue is in the fixative. Bryan ----- Original Message ----- From: "Bryan Watson" To: Sent: Thursday, May 07, 2009 10:49 AM Subject: [Histonet] pinning specimens for fixation We have a surgeon who likes to send us specimens that are pinned down to a board of some sort. He uses whatever makeshift board he can find and he wants us to order something for that purpose. Does anybody use anything like this something strong enough to hold a specimen pinned down without the pins being pulled out that is also disposable? If so, can you tell me what they're called and where to get them, thanks. I'll take any alternate ideas as well. Thanks, Bryan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cbrya <@t> lexclin.com Thu May 7 14:38:22 2009 From: cbrya <@t> lexclin.com (Carol Bryant) Date: Thu May 7 14:38:29 2009 Subject: [Histonet] disposal of body parts Message-ID: <37A1F9CAB9E21C41B39F3653B620D13E093BA204@exchange2003.lc.local> Hello Histonetters! I work in a small multispecialty clinic laboratory where the majority of our specimens are derm, gi biopsies, etc. Today we had an amputated finger sent for gross only from our ambulatory surgery center and the patient wishes to have it back. Does CAP have any specific requirements in regard to disposal of tissue that would apply here? Don't some patients wish to have their amputated limbs back for religious reasons? If so, what is the proper protocol? Should they sign some type of release form? Thanks, Carol Carol Bryant, CT (ASCP) Cytology/Histology Manager Pathology Services Lexington Clinic Phone (859) 258-4082 Fax (859) 258-4081 cbrya@lexclin.com NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by the State of Kentucky and/or Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient and have received this communication in error, please contact the sender and destroy all copies of the original message. Thank you. From AnthonyH <@t> chw.edu.au Thu May 7 17:40:20 2009 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu May 7 17:40:33 2009 Subject: [Histonet] NADPH-diaphorase In-Reply-To: <43B97B4C402C2C44AAA2A8D2C86A88B303A52F72@e2k3backend1.RCHSD.org> Message-ID: Eab, We purchase our NADH (reduced form) Cat No. N8129 from Sigma-aldrich Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Breisch, Eric Sent: Friday, 8 May 2009 2:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] NADPH-diaphorase Hi, hoping that someone might be able to share with us where they purchase NADPH-diaphorase for the NADPH stain. Thank you. EAB Eric A. Breisch, Ph.D. Clinical Anatomist Dept. of Pathology Rady Children's Hospital and Health Center Associate Clinical Professor of Anatomy Dept. of Surgery UCSD School of Medicine _______________________________________________ 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 note 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 lpwenk <@t> sbcglobal.net Thu May 7 21:00:07 2009 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Thu May 7 21:00:17 2009 Subject: [Histonet] Turnbull's Blue In-Reply-To: <5BF2F9FB24C0DC499CD3C4BB6F7B4481013E23FF@MAPMAIL02.bsci.bossci.com> Message-ID: <6969E9D3AAFD4B34881A86ED4EA904FB@HPPav2> Turnball blue (i.e., Schmorl stain) demonstrates argentaffinic components, such as melanin or argentaffin cells in the GI tract. Prussian blue (i.e., the iron stain) demonstrates iron (hemosiderin). What are you trying to demonstrate? Peggy A. Wenk, HTL(ASCP)SLS Beaumont Hospitals Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Wahlberg, Nikki Sent: Thursday, May 07, 2009 1:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Turnbull's Blue Is anyone performing the Turnbull's Blue Iron stain? If so, what type of control do you use. We are having a difficult time finding a good control in order to get the stain approved in our lab. Also, if anyone has some tips for the procedure they would be greatly appreciated. Thank you Nikki M Wahlberg Histotechnologist II Histo-Pathology Services Boston Scientific 763-694-5739 wahlbern@bsci.com CONFIDENTIALITY NOTICE: This e-mail transmission may contain confidential or legally privileged information that is intended only for the individual or entity named in the e-mail address. Any unauthorized distribution or copying of this transmittal or its attachments, if any, is prohibited. If you have received this e-mail transmission in error, please reply to the sender, so that Boston Scientific Corporation can arrange for proper delivery, then please delete the message from your inbox. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> waht.swest.nhs.uk Fri May 8 02:17:11 2009 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Fri May 8 02:17:17 2009 Subject: [Histonet] disposal of body parts Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06C0C6DB@wahtntex2.waht.swest.nhs.uk> Indeed they do; I believe some of the Asian Faiths require body parts to be buried with the person from whence they came. Certainly whilst I worked in London we had to 'return' an amputated leg to its rightful owner precisely for that reason. I also had to return a uterus, blocks and slides to a non-Asian lady for a reason I never quite understood. In the UK we are led to believe that body parts ought to be returned I suppose unless there is a health risk. In the case of the uterus I had to point out that it had been fixed and that the fixative including formalin (the COSHH sheet was issued); in the case of the leg I'm not quite sure. Once we had been directed to release the leg, which had also been fixed, I don't know where it went until its rightful owner 'threw off these mortal coils'. Years ago, of course, there were scandals over body parts being stored in plastic jars with brains and stuff in buckets. Some of those I gather were returned but don't actually know all the details. You should return them (they're not yours) but explain the health risks and try and suggest that they are better disposed of by the Lab. Foetal parts obviously are outside of this and I've always treated them as if they were people (as best as one can). If you detect foetal material in RPC then it ought to be removed and disposed of with care; we've sent these parts to the local crematorium for disposal and scattering on the land. We have returned foetuses to the parents for burial and we have buried them ourselves. Out of interest in retained products of conception in my last few jobs we didn't sample them as long as we were sure that they were foetal not molar. Help? Kemlo Rogerson e-mail kemlorogerson@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Carol Bryant Sent: 07 May 2009 20:38 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] disposal of body parts Hello Histonetters! I work in a small multispecialty clinic laboratory where the majority of our specimens are derm, gi biopsies, etc. Today we had an amputated finger sent for gross only from our ambulatory surgery center and the patient wishes to have it back. Does CAP have any specific requirements in regard to disposal of tissue that would apply here? Don't some patients wish to have their amputated limbs back for religious reasons? If so, what is the proper protocol? Should they sign some type of release form? Thanks, Carol Carol Bryant, CT (ASCP) Cytology/Histology Manager Pathology Services Lexington Clinic Phone (859) 258-4082 Fax (859) 258-4081 cbrya@lexclin.com NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by the State of Kentucky and/or Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient and have received this communication in error, please contact the sender 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 LINDA.MARGRAF <@t> childrens.com Fri May 8 08:16:27 2009 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri May 8 08:16:34 2009 Subject: [Histonet] how to unregister from histo-net In-Reply-To: <20090507111013.wkmxja2foc4gcggc@mail3.deltapathology.com> References: <20090507111013.wkmxja2foc4gcggc@mail3.deltapathology.com> Message-ID: <4A03EA5A.F783.00DA.0@childrens.com> Dear Kim (and other Histonet members): If you go to the website listed below(it is on the bottom of every Histonet message) and follow the instructions at the bottom of the page you can unsubscribe yourself. For future reference, you can use this site to subscribe, change your membership to the digest mode (with the daily messages all delivered as one batch every evening) etc. I am about to work on this list so I'll take you off this time. Take care, Linda M Histonet administrator >>> 5/7/2009 11:10 AM >>> Could anyone help me. I am trying to un-listg with the histo-net services. Please let me know. Thanks, Kim _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Please consider the environment before printing this e-mail

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From Valerie.T.Sailes.1 <@t> nd.edu Thu May 7 10:30:48 2009 From: Valerie.T.Sailes.1 <@t> nd.edu (Valerie Sailes) Date: Fri May 8 08:17:53 2009 Subject: [Histonet] In Situ Gelatin Zymography Message-ID: Hi, I'm looking for help with a new staining technique for my lab. We are hoping to perform In Situ Gelatin Zymography on some frozen section samples. We did a trial run and at first it appeared to have worked but looking at what should have been a negative we think either we have a lot of background autofluorescence or noise. I followed the protocol that I was given but we are not sure of what went wrong. Anyone out there familiar with this technique? Valerie Sailes Histology/Research Tech 400 FLSC vsailes@nd.edu From yanglf <@t> EVMS.EDU Thu May 7 11:03:21 2009 From: yanglf <@t> EVMS.EDU (Yang, Li) Date: Fri May 8 08:17:54 2009 Subject: [Histonet] fomalin-fixed tissue lysate Message-ID: Dear colleagues, I want to perform protein analysis (proteomics anylysis) from formalin-fixed tissue and have some questions to bother you. . 1. Although I carefully take the tissue off the wax block with knive, there is still some wax stick to the tissue. Anybody has good experience about how to dewax? 2. For tissue lysate, do I still need to do antigen retrievel like histochemictry? 3. Due to the crosslink by formalin, is the lysate still suitable for 2D gel analysis? Did Anybody try it? Any tips and suggestions are greatly appreciated. Best, lifang :: From joseph-galbraith <@t> uiowa.edu Fri May 8 08:19:45 2009 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Fri May 8 08:19:58 2009 Subject: [Histonet] FW: prover' pojaluista In-Reply-To: <766484.83301.qm@web65704.mail.ac4.yahoo.com> References: <766484.83301.qm@web65704.mail.ac4.yahoo.com> Message-ID: We have found that the key to IHC staining consistency following HIER (esp. when using a pressure cooker) is consistent and SLOW cool down. Opening the cooker too quickly can cause flash boiling and is dangerous but removing the slides from the cooker too rapidly and plunging them into room temp buffer can also be problematic. We found that careful trickling of DI water into the cooker for a consistent amount of time until the buffer reaches ambient temp to be effective. We actually time the cooling process and check the temp to be sure the slides are gently and adequately cooled before transferring to buffer prior to staining. Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Thursday, May 07, 2009 11:57 AM To: histonet@lists.utsouthwestern.edu; Margaryan, Naira Subject: Re: [Histonet] FW: prover' pojaluista Strictly speaking, IHC reactions do not depend on water temperature temperature. What you describe does not include dist. water between the IHC steps therefore the water temp. should not be affecting your reactions. If you use dist water after HIER you have to remember placing the slides?into room temp. buffer before starting the IHC procedure, so if there was any effect due to the dist. water, that should have take care of it. The inconsistency in your staining I think is not due to the dist. water temp. Ren? J. --- On Thu, 5/7/09, Margaryan, Naira wrote: From: Margaryan, Naira Subject: [Histonet] FW: prover' pojaluista To: "histonet@lists.utsouthwestern.edu" Date: Thursday, May 7, 2009, 11:23 AM Dear Histonetters, This is my question regarding IHC staining and water temperature: does IHC staining depend on water temperature in washing step? As always, I am using the same protocol for IHC. But one day it works perfectly, another day, on the same tissue, it does not work. Everything is same, solutions are fresh, and the only thing that is different is water temperature. I am using running distilled water before and after Antigen Retrieval step, after DAB, after counterstaining and after bluing reagent. Dd-water goes into autostainer: all blocking steps and after primary, secondary and tertiary. I am not using usual running water to exclude different Chlorine concentrations in different days. Have anybody of you ever paid attention to running distilled water temperature? What water temperature (cold or warm) you will recommend for washing steps? If this is critical, I can prepare my water up front. Hope to get an answer soon. Thanks, Naira _______________________________________________ Histonet mailing list Histonet@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 Nikki.Wahlberg <@t> bsci.com Fri May 8 08:31:01 2009 From: Nikki.Wahlberg <@t> bsci.com (Wahlberg, Nikki) Date: Fri May 8 08:31:05 2009 Subject: [Histonet] Turnbull's Blue In-Reply-To: <6969E9D3AAFD4B34881A86ED4EA904FB@HPPav2> References: <5BF2F9FB24C0DC499CD3C4BB6F7B4481013E23FF@MAPMAIL02.bsci.bossci.com> <6969E9D3AAFD4B34881A86ED4EA904FB@HPPav2> Message-ID: <5BF2F9FB24C0DC499CD3C4BB6F7B4481013E2403@MAPMAIL02.bsci.bossci.com> The study directors would like to use both the Prussian Blue and Turnbull's Blue to demonstrate two different types of iron in the tissue, ferric and ferrous. We are having trouble finding a control that contains both types of iron, thus we can not prove to the pathologist that are stain is indeed working. We in fact are not sure if the stain is working. Nikki M Wahlberg Histotechnologist II Histo-Pathology Services Boston Scientific 763-694-5739 wahlbern@bsci.com -----Original Message----- From: Lee & Peggy Wenk [mailto:lpwenk@sbcglobal.net] Sent: Thursday, May 07, 2009 9:00 PM To: Wahlberg, Nikki; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Turnbull's Blue Turnball blue (i.e., Schmorl stain) demonstrates argentaffinic components, such as melanin or argentaffin cells in the GI tract. Prussian blue (i.e., the iron stain) demonstrates iron (hemosiderin). What are you trying to demonstrate? Peggy A. Wenk, HTL(ASCP)SLS Beaumont Hospitals Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Wahlberg, Nikki Sent: Thursday, May 07, 2009 1:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Turnbull's Blue Is anyone performing the Turnbull's Blue Iron stain? If so, what type of control do you use. We are having a difficult time finding a good control in order to get the stain approved in our lab. Also, if anyone has some tips for the procedure they would be greatly appreciated. Thank you Nikki M Wahlberg Histotechnologist II Histo-Pathology Services Boston Scientific 763-694-5739 wahlbern@bsci.com CONFIDENTIALITY NOTICE: This e-mail transmission may contain confidential or legally privileged information that is intended only for the individual or entity named in the e-mail address. Any unauthorized distribution or copying of this transmittal or its attachments, if any, is prohibited. If you have received this e-mail transmission in error, please reply to the sender, so that Boston Scientific Corporation can arrange for proper delivery, then please delete the message from your inbox. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joseph-galbraith <@t> uiowa.edu Fri May 8 08:59:55 2009 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Fri May 8 09:00:02 2009 Subject: [Histonet] disposal of body parts In-Reply-To: <37A1F9CAB9E21C41B39F3653B620D13E093BA204@exchange2003.lc.local> References: <37A1F9CAB9E21C41B39F3653B620D13E093BA204@exchange2003.lc.local> Message-ID: Carol: Native Americans also generally have a belief that body parts must be buried with the rest of the body at the time of death and will recover any severed limbs for temporary burial until the person passes. It is critical to have a release form that includes warnings regarding the hazards of handling human remains (biohazards and formaldehyde hazards for example) as well as an admonition that the recipient must follow all federal, state and local regulations that may apply (both in your state and the destination state if different - you do not have to spell out the regulations - just warn the patient that they must comply). As a further precaution, we do not release body parts directly to the patient or family, we release to the patient's in-house clinician who then reviews the warnings with the patient before releasing. Both the clinician and the patient sign the release form which then becomes part of the patient's records and a copy is retained in the lab for our records. Someone mentioned releasing fetal remains for burial. Be very careful here as the regulations vary greatly from state to state and sometimes even jurisdiction to jurisdiction with a state. In our state a properly executed fetal death certificate must be filed with the county of death and a burial transit permit issued whenever the fetus is more than 20 weeks of gestation before the remains can be transported by anyone other than a licensed funeral director. Have your institutional legal counsel check for your local regulation before proceeding. Good luck. Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Carol Bryant Sent: Thursday, May 07, 2009 2:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] disposal of body parts Hello Histonetters! I work in a small multispecialty clinic laboratory where the majority of our specimens are derm, gi biopsies, etc. Today we had an amputated finger sent for gross only from our ambulatory surgery center and the patient wishes to have it back. Does CAP have any specific requirements in regard to disposal of tissue that would apply here? Don't some patients wish to have their amputated limbs back for religious reasons? If so, what is the proper protocol? Should they sign some type of release form? Thanks, Carol Carol Bryant, CT (ASCP) Cytology/Histology Manager Pathology Services Lexington Clinic Phone (859) 258-4082 Fax (859) 258-4081 cbrya@lexclin.com NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by the State of Kentucky and/or Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient and have received this communication in error, please contact the sender 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 llewllew <@t> shaw.ca Fri May 8 09:35:25 2009 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri May 8 09:36:29 2009 Subject: [Histonet] Turnbull's Blue In-Reply-To: <5BF2F9FB24C0DC499CD3C4BB6F7B4481013E2403@MAPMAIL02.bsci.bossci.com> References: <5BF2F9FB24C0DC499CD3C4BB6F7B4481013E23FF@MAPMAIL02.bsci.bossci.com> <6969E9D3AAFD4B34881A86ED4EA904FB@HPPav2> <5BF2F9FB24C0DC499CD3C4BB6F7B4481013E2403@MAPMAIL02.bsci.bossci.com> Message-ID: <7BF9E721D36F4810AAEBDD8A3078B1DE@BryanPC> It was my understanding that there is not usually any ferrous iron that can be demonstrated in tissue, any of it present being securely bound up and masked in larger molecules. The method used (Tirmann Schmelzer) simply replaces the potassium ferrocyanide of Perls' method with potassium ferricyanide, and ferrous iron then forms Turnbull's blue. The control suggested is lung tissue from a miner who has inhaled iron ore dust, so both ferrous and ferric compounds would be expected to be present. Where you would get that I haven't a clue. Possibly you could make one by injecting a ferrous salt solution followed by a ferric salt solution into fresh lung, then fixing it. I suspect that a natural control will be difficult to come by. Bryan Llewellyn ----- Original Message ----- From: "Wahlberg, Nikki" To: Sent: Friday, May 08, 2009 6:31 AM Subject: RE: [Histonet] Turnbull's Blue The study directors would like to use both the Prussian Blue and Turnbull's Blue to demonstrate two different types of iron in the tissue, ferric and ferrous. We are having trouble finding a control that contains both types of iron, thus we can not prove to the pathologist that are stain is indeed working. We in fact are not sure if the stain is working. Nikki M Wahlberg Histotechnologist II Histo-Pathology Services Boston Scientific 763-694-5739 wahlbern@bsci.com -----Original Message----- From: Lee & Peggy Wenk [mailto:lpwenk@sbcglobal.net] Sent: Thursday, May 07, 2009 9:00 PM To: Wahlberg, Nikki; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Turnbull's Blue Turnball blue (i.e., Schmorl stain) demonstrates argentaffinic components, such as melanin or argentaffin cells in the GI tract. Prussian blue (i.e., the iron stain) demonstrates iron (hemosiderin). What are you trying to demonstrate? Peggy A. Wenk, HTL(ASCP)SLS Beaumont Hospitals Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Wahlberg, Nikki Sent: Thursday, May 07, 2009 1:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Turnbull's Blue Is anyone performing the Turnbull's Blue Iron stain? If so, what type of control do you use. We are having a difficult time finding a good control in order to get the stain approved in our lab. Also, if anyone has some tips for the procedure they would be greatly appreciated. Thank you Nikki M Wahlberg Histotechnologist II Histo-Pathology Services Boston Scientific 763-694-5739 wahlbern@bsci.com CONFIDENTIALITY NOTICE: This e-mail transmission may contain confidential or legally privileged information that is intended only for the individual or entity named in the e-mail address. Any unauthorized distribution or copying of this transmittal or its attachments, if any, is prohibited. If you have received this e-mail transmission in error, please reply to the sender, so that Boston Scientific Corporation can arrange for proper delivery, then please delete the message from your inbox. _______________________________________________ Histonet mailing list Histonet@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 putnamj <@t> ggclinic.com Fri May 8 09:44:57 2009 From: putnamj <@t> ggclinic.com (Putnam, Jodi) Date: Fri May 8 09:45:06 2009 Subject: [Histonet] Prostate bx processing time and fixation time Message-ID: Hi I have a question for everyone today. I have recently taken on prostate bx's as I have said in other postings. I had to adjust my processing time and think that's going much better. My question is, today at 10:45 am they are doing a prostate bx (12 specimens) on a patient and my pathologist would like it processed immediately upon receipt. I was wondering, with the size of the prostate biopsies, what is a standard fixation time prior to processing. I don't want to have the biopsy done at 10:45 and put them on the processor immediately for fear of them not being fixed. I am sure that the fixation time is not long as compared to my derm specimens, but what is a good standard to follow as far as fixation time on prostates prior to processing. Ideally this is to get the case out the same day but I don't want to process unfixed specimens and cause serious problems. I took out the formalin steps on the short processing run that I have set up on my Excelsior. Usually I receive the prostate case in the afternoon and gross it in, let is sit overnight and then run them as soon as I come in the next day. Any help would be appreciated. I do not want to cut corners and jeopardize specimen quality. So, what is the minimum time that you think is fair for fixation prior to me processing them today? It's Friday and I don't want to leave loose ends but also don't want to compromise the specimen quality. HELP!!!! :) Thanks so much for all of the help and advise that everyone has given me. This site is wonderful and has helped me in so many ways. I will anxiously await the replies. Its 9:43 am here so I have one hour to make a decision. :) Happy Friday :) (I hope.......) Jodi Putnam (HT,ASCP) Graves Gilbert Clinic Pathology Department 201 Park Street Bowling Green, KY 42102 (270) 393-2728 (voicemail) (270) 393-2795 Fax : (270) 393-2736 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 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 have received this email in error please notify the originator of the message. This footer also confirms that this email message has been scanned for the presence of computer viruses. Any views expressed in this message are those of the individual sender, except where the sender specifies and with authority, states them to be the views of Graves-Gilbert Clinic. Scanning of this message and addition of this footer is performed by Websense Email Security software in conjunction with virus detection software. From jcampbell <@t> vdxpathology.com Fri May 8 11:11:30 2009 From: jcampbell <@t> vdxpathology.com (Jennifer Campbell) Date: Fri May 8 11:11:36 2009 Subject: [Histonet] Toulidine blue stain Message-ID: <5658CBDB9EAE6545ABE50D2563D81BF821CEE2@VDXSERVER01.vdxpathology.local> Hi everyone, I'm trying to find a protocol for a toulidine blue stain for paraffin sections. I've looked in different text books and online and can't seem to find anything helpful. Does anyone have a good protocol for this? I need to order the stain rather soon but, first I want to know how much I'm going to need to run the stain. Thanks in advance, Jen C. From Kjones <@t> upei.ca Fri May 8 12:14:27 2009 From: Kjones <@t> upei.ca (Kathleen Jones) Date: Fri May 8 12:14:39 2009 Subject: [Histonet] t blue Message-ID: <4A043E44.BDAE.008B.0@groupwise.upei.ca> Hi Jennifer, I forgot to mention two important steps to a successful T. Blue stain...Always mix the t.blue solution very well just prior to use and make sure your alcohol, particularly the 95% is fresh. It gives great results. Have a great weekend Kathy From akbitting <@t> geisinger.edu Fri May 8 12:46:58 2009 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri May 8 12:47:07 2009 Subject: [Histonet] SMMS Message-ID: <4A0437D2.2B7F.00C9.0@geisinger.edu> We've been having trouble getting satisfactory staining for SMMS on our BenchmarkXTs. Is anyone getting a good result running these on the XT platform. If so, would you mind sharing your secret?? Thanks, Angie IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From syedj <@t> wyeth.com Fri May 8 13:09:24 2009 From: syedj <@t> wyeth.com (Jameel Syed) Date: Fri May 8 13:09:44 2009 Subject: [Histonet] Frozen placenta (Human, Cynomolgus monkey & Marmoset) Message-ID: <4A043D14020000F800017A61@gvl011m.gv.us.pri.wyeth.com> Hi all, Does anybody know of a good US commercial source for snap frozen placenta (Human, Cynomolgus monkey & Marmoset). Ideally, I am looking for samples that are OCT embedded and snap frozen. I have already checked with Asterand, Alphagenesis, Biomax, Bioserve and Lifespan. Thanks, Jameel Jameel Syed, BS, HT/HTL, QIHC (ASCP) Senior Research Scientist I Exploratory Drug Safety Wyeth Research 1 Burtt Road, Andover, MA 01810 Tel: 978-247-1348 Fax: 978-684-4248 syedj@wyeth.com From alyssa <@t> alliedsearchpartners.com Fri May 8 13:58:23 2009 From: alyssa <@t> alliedsearchpartners.com (Alyssa Peterson) Date: Fri May 8 13:58:27 2009 Subject: [Histonet] Histology Position Long Island, NY Message-ID: Hot Job For Talented Histotechnologist * **Please review the Job Description and call me immediately to schedule an interview as this job will not last long based on the competitive benefits and competitive pay.* *Please review the job description and send an updated resume with any other questions to Alyssa@alliedsearchpartners.com * *If you are not interested but you know someone who is, we offer $1000 as a referral bonus if we place one of your referrals in a position. Please forward this email to anyone fit for this position. * *If this is not the job for you, but you are currently looking for a position, please send me a description of what you are looking for and your updated resume, as we have positions in your area!* Location: Long Island: *Shirly**, **NY* FULL TIME HISTOTECHNOLOGIST 3-5 years experience as a Histotechnologist, with at least 1-2 years in leadership role preferred and knowledge/ experience in immunohistochemistry is a plus. Benefit package includes moving expences, healthcare coverage, 401K with profit sharing plan,CME expences and life insurance. Come work with a growing private pathology laboratory in Suffolk County in Long Island, NY established in 1998. We work as a team in a friendly and professional environment to provide highest quality service to our client. We use state of art pathology equipments. We have excellent compensation package based on experience and productivity. Please apply by sending resume to: Alyssa@alliedsearchpartners.com -- Alyssa Peterson Allied Search Partners O: 770.621.2639 ext. 4 F: 770.621.2640 From loftonjt <@t> holycrosshealth.org Fri May 8 14:43:26 2009 From: loftonjt <@t> holycrosshealth.org (Jimmy Lofton) Date: Fri May 8 14:43:42 2009 Subject: [Histonet] Liver biopsy concerns Message-ID: <4A04531D.9B70.0056.0@holycrosshealth.org> Has anyone had any concerns regarding the samples of Liver Biopsies from Ultra Sound that uses any anesthesia at the injection site? Air drying artifact, autolysis and shattering of the specimen. Thanks, Jimmy Jimmy Lofton, M.S., HT,CT(ASCP) Manager Histology Laboratory Holy Cross Hospital 1500 Forest Glen Road Silver Spring, MD 20910-1484 301-754-7353 (Phone) 301-754-8563 (Fax) loftonjt@holycrosshealth.org Trinity Health MailGate made the following annotations --------------------------------------------------------------------- CONFIDENTIALITY NOTICE: This e-mail, including any attachments, is for the sole use of the intended recipient and may contain information that is confidential and privileged under state and Federal privacy laws. If you received this e-mail in error, be aware that any unauthorized use; disclosure, copying, or distribution is strictly prohibited. Please contact the sender immediately and destroy all copies of this message. --------------------------------------------------------------------- From cbarone <@t> NEMOURS.ORG Fri May 8 14:56:57 2009 From: cbarone <@t> NEMOURS.ORG (Barone, Carol ) Date: Fri May 8 14:56:58 2009 Subject: [Histonet] Virginia Beach Meeting Message-ID: <37E4BAC017F57141AF64FAA5AEB04CE8033A728F@wlmmsx01.nemours.org> Learn, Earn, Eat and Network! Perhaps even find those new or better jobs! ...or, attend the "Open Forum"...and bring your questions to discuss with other tech's. We couldn't make it any easier or less expensive for you to Learn and Earn those needed CEU's... May 12th is the last day to pre-register for the Virginia Beach "Learn and Earn - Saturday Seminars", hosted by the regional sister states of DE, MD. PA and NJ in support of re-activating the Virginia State Society. 2 excellent workshops, 5 CEU's, 1 continental breakfast and a free lunch.... with every pre-registration by May 12th for full day attendees. $10 registration fee will be waived for all attendees who pre-register for both sessions by may 12th and a $5 discount will be given on each seminar to attendees with state memberships in DE, MD, PA and NJ...or... with a new, or renewed, membership in the Virginia Society. You can earn 5 CEU's with as little as $60...and get a free breakfast and lunch! So why are you staying home on May 16th? Register now!!! This is how every state society supports it's sister states....and tech's! Seminars: Antibody Panels in the Anatomic Pathology Lab Multiplex Stains for the Anatomic Pathology Lab For information on how to register: cbarone@nemours.org PS: Education is professional development! You can go to the beach on Sunday! See you there! From dcrippen <@t> buckinstitute.org Fri May 8 15:41:44 2009 From: dcrippen <@t> buckinstitute.org (Danielle Crippen) Date: Fri May 8 15:41:49 2009 Subject: [Histonet] Paraffin processing schedule-Cynamologus monkey tibia Message-ID: <69142A827D247F45877B305E991DB61F18E28B@inverness.buckcenter.org> Dear Histo-experts, Can you share your processing schedules for monkey (or human) bone samples. I need to paraffin embed a cynamologus monkey tibia. It's decalcified with the endpoint confirmed (with ammonium oxalate). I've had good success with mouse bones with the following schedule, but am not sure it's long enough for this larger sample: - 70% EtOH 1.5 hours - 80% EtOH 1.5 hours - 95% EtOH 2x 1.5 hours - 100% EtOH 2x 1.5 hours - Clearite 3 2x1.5 hours - Tissue Prep 2 paraffin 4x1.5 hours 60 degrees under vacuum - Embed in tissue prep 2 We're hand processing, so any/all advice is welcome!! Many thanks in advance! d Danielle Crippen Morphology and Imaging Core Manager Buck Institute for Age Research From stamptrain <@t> yahoo.com Fri May 8 17:13:10 2009 From: stamptrain <@t> yahoo.com (Roger Moretz) Date: Fri May 8 17:13:12 2009 Subject: [Histonet] Paraffin processing schedule-Cynamologus monkey tibia In-Reply-To: <69142A827D247F45877B305E991DB61F18E28B@inverness.buckcenter.org> References: <69142A827D247F45877B305E991DB61F18E28B@inverness.buckcenter.org> Message-ID: <318207.26632.qm@web55806.mail.re3.yahoo.com> Your schedule should be fine for any large tissue, and actually seems a bit overlong for mouse.? But if it works, it works. Roger Moretz, ret. ----- Original Message ---- From: Danielle Crippen To: histonet@lists.utsouthwestern.edu Sent: Friday, May 8, 2009 4:41:44 PM Subject: [Histonet] Paraffin processing schedule-Cynamologus monkey tibia Dear Histo-experts, Can you share your processing schedules for monkey (or human) bone samples.? I need to paraffin embed a cynamologus monkey tibia.? It's decalcified with the endpoint confirmed (with ammonium oxalate).? I've had good success with mouse bones with the following schedule, but am not sure it's long enough for this larger sample: -? ? ? ? 70% EtOH 1.5 hours -? ? ? ? 80% EtOH 1.5 hours -? ? ? ? 95% EtOH 2x 1.5 hours -? ? ? ? 100% EtOH 2x 1.5 hours -? ? ? ? Clearite 3 2x1.5 hours -? ? ? ? Tissue Prep 2 paraffin 4x1.5 hours 60 degrees under vacuum -? ? ? ? Embed in tissue prep 2 We're hand processing, so any/all advice is welcome!! Many thanks in advance! d Danielle Crippen Morphology and Imaging Core Manager Buck Institute for Age Research _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From amosbrooks <@t> gmail.com Fri May 8 17:24:17 2009 From: amosbrooks <@t> gmail.com (Amos Brooks) Date: Fri May 8 17:24:21 2009 Subject: [Histonet] Toulidine blue stain Message-ID: <582736990905081524n53396db7ucf8df74f19b0e3d2@mail.gmail.com> Hi, I've tried this procedure and it works very well, as do most of those found on this site. http://www.ihcworld.com/_protocols/special_stains/toluidine_blue.htm (Nice work those of you that put this site togetherby the way!) Good luck, Amos Date: Fri, 8 May 2009 09:11:30 -0700 From: "Jennifer Campbell" Subject: [Histonet] Toulidine blue stain To: Message-ID: <5658CBDB9EAE6545ABE50D2563D81BF821CEE2@VDXSERVER01. vdxpathology.local> Content-Type: text/plain; charset="US-ASCII" Hi everyone, I'm trying to find a protocol for a toulidine blue stain for paraffin sections. I've looked in different text books and online and can't seem to find anything helpful. Does anyone have a good protocol for this? I need to order the stain rather soon but, first I want to know how much I'm going to need to run the stain. Thanks in advance, Jen C. From ccrowder <@t> vetmed.lsu.edu Fri May 8 22:56:26 2009 From: ccrowder <@t> vetmed.lsu.edu (Cheryl Crowder) Date: Fri May 8 22:57:30 2009 Subject: [Histonet] Toluidine Blue stain Message-ID: Jen - We do T=blue staining almost daily. If you will contact me directly I will send you our directions. It is a very inexpensive stain, and can be reused. You will need to pH it often. 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 gu.lang <@t> gmx.at Sat May 9 13:34:39 2009 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat May 9 13:34:51 2009 Subject: [Histonet] KI67 test for fixation? Message-ID: <8E55BCF760E745548BEF92830AE0FA54@dielangs.at> Hi all! Can anybody explain the correlation of KI-67 stainability and the status of formaldehyde-fixation? I cannot remember, if an under- or overfixed tissue shows the positive KI67 nuclei. And I don't remember, if this test is still valuable. Please help, Gudrun From dermpathsy <@t> gmail.com Sat May 9 14:16:46 2009 From: dermpathsy <@t> gmail.com (Sate Hamza) Date: Sat May 9 14:16:50 2009 Subject: [Histonet] Human herpesvirus 7 Message-ID: <8854ff80905091216p291f46d9nbe5db84e9e16cbc5@mail.gmail.com> Dear Histonetters, A colleague dermatopathologist posted the inquiry below on another forum regarding immunohistochemistry for human herpesvirus 7. Does anybody know a place that does this? "Does anyone have the ability to check for HH7 on paraffin blocks (for pityriasis rosea) as a routine IHC test?" Thank you in advance for any information. Sate -- Sate Hamza, MD, FRCPC Dermatopathologist Winnipeg, Canada From c.m.vanderloos <@t> amc.uva.nl Mon May 11 01:34:27 2009 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Mon May 11 01:34:38 2009 Subject: [Histonet] RE: KI67 test for fixation? Message-ID: Gudrun,Ki67 as stained by SP6 rabbit monoclonal antibody survives quite long formaldehyde fixation. Up to 2 months of fixation, the staining intensity is as strong as after 1 day of formaldehyde fixation. Even after one year in formalin there is still staining of this marker. I don't know what the status of Ki67 will be after too short fixation. Cheers,ChrisChris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The NetherlandsDate: Sat, 9 May 2009 20:34:39 +0200 From: "Gudrun Lang" Subject: [Histonet] KI67 test for fixation? To: histonet@lists.utsouthwestern.edu Hi all! Can anybody explain the correlation of KI-67 stainability and the status of formaldehyde-fixation? I cannot remember, if an under- or overfixed tissue shows the positive KI67 nuclei. And I don't remember, if this test is still valuable. Please help, Gudrun From annigyg <@t> gmail.com Mon May 11 02:02:26 2009 From: annigyg <@t> gmail.com (Anne van Binsbergen) Date: Mon May 11 02:02:33 2009 Subject: [Histonet] Liver biopsy concerns In-Reply-To: <4A04531D.9B70.0056.0@holycrosshealth.org> References: <4A04531D.9B70.0056.0@holycrosshealth.org> Message-ID: We have mysterious artefact in our Renal biopsies (shattering and strange cellular 'cracking') which i am pretty sure come from a similar scenario the pathologists are dead keen to blame my techs - but i have conducted several detailed investigations of all our processes and found NOTHING lab-related to convince me that they are to blame have been on several procedures myself and am very suspicious of the interventional radiologists and particularly the local anaesthetic being pumped into the op-site but alas, no-one listens - they just carry on and do their own thing 2009/5/8 Jimmy Lofton > Has anyone had any concerns regarding the samples of Liver Biopsies from > Ultra Sound that uses any anesthesia at the injection site? Air drying > artifact, autolysis and shattering of the specimen. > > Thanks, > > Jimmy > > > Jimmy Lofton, M.S., HT,CT(ASCP) > Manager Histology Laboratory > Holy Cross Hospital > 1500 Forest Glen Road > Silver Spring, MD 20910-1484 > 301-754-7353 (Phone) > 301-754-8563 (Fax) > loftonjt@holycrosshealth.org > > > Trinity Health MailGate made the following annotations > --------------------------------------------------------------------- > CONFIDENTIALITY NOTICE: This e-mail, including any attachments, is for the > sole use of the intended recipient and may contain information that is > confidential and privileged under state and Federal privacy laws. If you > received this e-mail in error, be aware that any unauthorized use; > disclosure, copying, or distribution is strictly prohibited. Please contact > the sender immediately and destroy all copies of this message. > --------------------------------------------------------------------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Anne van Binsbergen (Hope) Abu Dhabi UAE From Kemlo.Rogerson <@t> waht.swest.nhs.uk Mon May 11 02:09:26 2009 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon May 11 02:09:31 2009 Subject: [Histonet] Liver biopsy concerns Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06C0C7C7@wahtntex2.waht.swest.nhs.uk> Why don't you do a scrimp slide? Just drag the biopsy over a clean glass slide to dislodge as many cells as possible and then fix the cells on the slide with spray fix. You stain the cells and if they are similarly disrupted it isn't your Techs; the problem is therefore in vitro. Kemlo Rogerson e-mail kemlorogerson@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne van Binsbergen Sent: 11 May 2009 08:02 To: Jimmy Lofton Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Liver biopsy concerns We have mysterious artefact in our Renal biopsies (shattering and strange cellular 'cracking') which i am pretty sure come from a similar scenario the pathologists are dead keen to blame my techs - but i have conducted several detailed investigations of all our processes and found NOTHING lab-related to convince me that they are to blame have been on several procedures myself and am very suspicious of the interventional radiologists and particularly the local anaesthetic being pumped into the op-site but alas, no-one listens - they just carry on and do their own thing 2009/5/8 Jimmy Lofton > Has anyone had any concerns regarding the samples of Liver Biopsies > from Ultra Sound that uses any anesthesia at the injection site? Air > drying artifact, autolysis and shattering of the specimen. > > Thanks, > > Jimmy > > > Jimmy Lofton, M.S., HT,CT(ASCP) > Manager Histology Laboratory > Holy Cross Hospital > 1500 Forest Glen Road > Silver Spring, MD 20910-1484 > 301-754-7353 (Phone) > 301-754-8563 (Fax) > loftonjt@holycrosshealth.org > > > Trinity Health MailGate made the following annotations > --------------------------------------------------------------------- > CONFIDENTIALITY NOTICE: This e-mail, including any attachments, is for > the sole use of the intended recipient and may contain information > that is confidential and privileged under state and Federal privacy > laws. If you received this e-mail in error, be aware that any > unauthorized use; disclosure, copying, or distribution is strictly > prohibited. Please contact the sender immediately and destroy all copies of this message. > --------------------------------------------------------------------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Anne van Binsbergen (Hope) Abu Dhabi UAE _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tifei <@t> foxmail.com Mon May 11 06:41:51 2009 From: tifei <@t> foxmail.com (TF) Date: Mon May 11 06:42:09 2009 Subject: [Histonet] Anti -rat CD31 Message-ID: <200905111941463478449@foxmail.com> Dear All Just wonder whether you know any anti-rat CD31 that work on PFA-fixed tissues - frozen section ? It seems all of them just work on acetone fixed tissues! The tissue quality can not be promised ! Even you try antigen retrieval, it wont work? 2009-05-11 TF From tifei <@t> foxmail.com Mon May 11 08:43:36 2009 From: tifei <@t> foxmail.com (TF) Date: Mon May 11 08:43:54 2009 Subject: [Histonet] 0.2M borate acid pH 7.0 Message-ID: <200905112143309190095@foxmail.com> My recipe for 0.2M borate acid is: Sodium Borate acid.10 H2O (molecular weight 381.42, Na4B4O4.10H2O) 19.07 gram H2O 1 Litre Adjust pH = 7.0 with 5M HCl Is this fine? 2009-05-11 TF From anh2006 <@t> med.cornell.edu Mon May 11 08:56:26 2009 From: anh2006 <@t> med.cornell.edu (anh2006@med.cornell.edu) Date: Mon May 11 08:59:15 2009 Subject: [Histonet] Anti -rat CD31 Message-ID: <820446333-1242050347-cardhu_decombobulator_blackberry.rim.net-737702447-@bxe1087.bisx.prod.on.blackberry> Clone Mec13.3 (BD Pharmingen sells it among others) works great on PFA fixed frozen tissues but you must use trypsin as an antigen retrieval step. I do 30 min at 37 deg C. Depending on your tissue that may need some adjusting. ------Original Message------ From: TF Sender: histonet-bounces@lists.utsouthwestern.edu To: histonet ReplyTo: tifei@foxmail.com Sent: May 11, 2009 7:41 AM Subject: [Histonet] Anti -rat CD31 Dear All Just wonder whether you know any anti-rat CD31 that work on PFA-fixed tissues - frozen section ? It seems all of them just work on acetone fixed tissues! The tissue quality can not be promised ! Even you try antigen retrieval, it wont work? 2009-05-11 TF _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Mon May 11 09:52:27 2009 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Mon May 11 09:52:36 2009 Subject: [Histonet] rush kidney biopsies Message-ID: Hi all! Until now we were lucky to have no short-processing-protocols to deal with. Now we start with transplantation kidney biopsies and a two-hour-protocol on a VIP. Receiving biopsy ? 30 min NBF ? then VIP: 30 min NBF 40?C, 15 min 70% alk, 15 min 80% alk, 15 min 96% alk, 15 min 100% alk, 30 min paraffin We do HE and specialstains ( Jones, PAS, EvG, Congored, Trichrom SFOG) and immunofluorescenc (Ig) and immunohistochemistry (C4d, HLA, Polyomavirus). If anyone have bad experiences with the short protocols, would you mind to share those with me? Regarding cutting and staining. What are the points, I have to take care of? Good experiences are also welcome. Bye Gudrun From Rcartun <@t> harthosp.org Mon May 11 10:33:21 2009 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Mon May 11 10:33:34 2009 Subject: [Histonet] rush kidney biopsies In-Reply-To: References: Message-ID: <4A080D00.7400.0077.1@harthosp.org> Yes, we have had problems with our IP stains (primarily C4d) when performed on rushed transplant biopsies. The paraffin-embedded tissue does not hold up well to enzyme digestion when it's not properly fixed. After educating our transplant surgeons they are perfectly happy to wait until the next day. In situations where an immediate read is needed, two cores can be taken; one for rush morphology and one for overnight fixation and subsequent immunostaining. Richard Richard W. Cartun, Ph.D. Director, Histology & Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Gudrun Lang" 5/11/2009 10:52 AM >>> Hi all! Until now we were lucky to have no short-processing-protocols to deal with. Now we start with transplantation kidney biopsies and a two-hour-protocol on a VIP. Receiving biopsy ? 30 min NBF ? then VIP: 30 min NBF 40?C, 15 min 70% alk, 15 min 80% alk, 15 min 96% alk, 15 min 100% alk, 30 min paraffin We do HE and specialstains ( Jones, PAS, EvG, Congored, Trichrom SFOG) and immunofluorescenc (Ig) and immunohistochemistry (C4d, HLA, Polyomavirus). If anyone have bad experiences with the short protocols, would you mind to share those with me? Regarding cutting and staining. What are the points, I have to take care of? Good experiences are also welcome. Bye Gudrun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Mon May 11 10:52:55 2009 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Mon May 11 10:53:03 2009 Subject: AW: [Histonet] rush kidney biopsies In-Reply-To: <4A080D00.7400.0077.1@harthosp.org> References: <4A080D00.7400.0077.1@harthosp.org> Message-ID: <170DEF69E5F94C2380CA7FB4AFD4520E@dielangs.at> Thank you Richard for the input. Our C4d protocol is with HIER pH 9 (CC1, Ventana). I will have a look at it, maybe the retrieval step has to be shortened. Everybody knows, that the outcome isn't worth the effort, but nobody wants to disappoint the new clinician. Best regards Gudrun -----Urspr?ngliche Nachricht----- Von: Richard Cartun [mailto:Rcartun@harthosp.org] Gesendet: Montag, 11. Mai 2009 17:33 An: gu.lang@gmx.at; histonet@lists.utsouthwestern.edu Betreff: Re: [Histonet] rush kidney biopsies Yes, we have had problems with our IP stains (primarily C4d) when performed on rushed transplant biopsies. The paraffin-embedded tissue does not hold up well to enzyme digestion when it's not properly fixed. After educating our transplant surgeons they are perfectly happy to wait until the next day. In situations where an immediate read is needed, two cores can be taken; one for rush morphology and one for overnight fixation and subsequent immunostaining. Richard Richard W. Cartun, Ph.D. Director, Histology & Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Gudrun Lang" 5/11/2009 10:52 AM >>> Hi all! Until now we were lucky to have no short-processing-protocols to deal with. Now we start with transplantation kidney biopsies and a two-hour-protocol on a VIP. Receiving biopsy ? 30 min NBF ? then VIP: 30 min NBF 40?C, 15 min 70% alk, 15 min 80% alk, 15 min 96% alk, 15 min 100% alk, 30 min paraffin We do HE and specialstains ( Jones, PAS, EvG, Congored, Trichrom SFOG) and immunofluorescenc (Ig) and immunohistochemistry (C4d, HLA, Polyomavirus). If anyone have bad experiences with the short protocols, would you mind to share those with me? Regarding cutting and staining. What are the points, I have to take care of? Good experiences are also welcome. Bye Gudrun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From smcbride <@t> cs.cmu.edu Mon May 11 12:07:14 2009 From: smcbride <@t> cs.cmu.edu (Sean McBride) Date: Mon May 11 12:07:24 2009 Subject: [Histonet] Request for Owner's Manual: Leica SM2500E Message-ID: Colleagues, I have a Leica SM2500E sliding sledge microtome for which I am searching for a copy of the owner's manual. If anyone has one that they could share with me, I would be most indebted. Best regards, ~Sean McBride Senior Researcher Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-901-7540 (m) 412-268-8641 (fax) smcbride@cs.cmu.edu From estellamireles <@t> gmail.com Mon May 11 12:56:29 2009 From: estellamireles <@t> gmail.com (Stella Mireles) Date: Mon May 11 12:56:33 2009 Subject: [Histonet] Phosphofuctokinase Staining Procedure Message-ID: Would anyone be willing to send me their procedure for Phosphofuctokinase, or a site where I can pull this up myself. The reference book that I was supplied with it not enough for me to pull what I need. Thanks From gayle.callis <@t> bresnan.net Mon May 11 13:01:30 2009 From: gayle.callis <@t> bresnan.net (gayle callis) Date: Mon May 11 13:01:49 2009 Subject: [Histonet] Re: Direct conjugates for immunofluorescence staining Message-ID: <000401c9d262$84927af0$8db770d0$@callis@bresnan.net> Jennifer, You Wrote: I work with a scientist who insists on using primary antibodies directly conjugated with FITC or Texas Red for IHC. In my past experience these directly conjugated antibodies didn't give a strong enough signal for use in IHC (I've seen them used only for FACS analysis). I would appreciate your professional comments. Jennifer M. Anderson, Scientist Halozyme Therapeutics, Inc. 11404 Sorrento Valley Road San Diego, CA 92121 858-704-8333 If your antibody is conjugated to a fluorophore, as you described, you may run into the problem where the FITC and Texas Red molecules are in too close proximation to each other and will "quench" due to a trading of electrons between the FITC molecules when these are excited. This is a problem we had with murine CD4-FITC and murine CD8-FITC. The result is the antibody binds to the antigen, a protein, but the quenching causes the FITC to not fluoresce. Quenching is a NOT photobleaching aka "fading", but a different physical chemical happening that "reduces the excited state lifetime, and the quantum yield of the affected fluorophore." You can go to www.olumpusconfocal.com/theory/fluorphoresintro.html and read about this under Quenching and Photobleaching (two different things). After long discussions with my Physical Chemist husband who works with quenching studies on fluorescent molecules, we do not use direct fluorophore conjugated antibodies in the lab for staining tissue sections anymore. The Jablonski Diagram also shows this. It will not happen with all antibodies that are conjugated to fluorophores due to their spatial relationships (in terms of nanometers) but if it does, Ray Koellings suggestion to come back with an antiFITC or Anti Texas red should solve the problem. We are better served by having the primary antibodies biotinylated, then come back with a Streptavidin Alexa fluor dye (488 for FITC, and 594 for Texas Red). From Dorothy.L.Webb <@t> HealthPartners.Com Tue May 12 09:48:07 2009 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Tue May 12 09:48:16 2009 Subject: [Histonet] General data cassette printer Message-ID: <65365F35C0F2EF4D846EC3CA73E49C43682629347A@HPEMX3.HealthPartners.int> Is anyone in Histoland using the General Data cassette printer or trying it out?? If so, please let me know your thoughts on the printer and process!! Much appreciated!! Dorothy Webb, HT Regions Histology Technical Supervisor 651-254-2962 ________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 From eca9 <@t> georgetown.edu Tue May 12 10:44:27 2009 From: eca9 <@t> georgetown.edu (Eva Permaul) Date: Tue May 12 10:44:30 2009 Subject: [Histonet] EGFR antibody for staining mouse tissue? Message-ID: <4A09995B.9040607@georgetown.edu> Hello, Does anybody use an EGFR antibody for staining on mouse tissue? Is it a published antibody? Would you be willing to share your protocol? Thanks, Eva Georgetown University From cbarone <@t> NEMOURS.ORG Tue May 12 11:34:59 2009 From: cbarone <@t> NEMOURS.ORG (Barone, Carol ) Date: Tue May 12 11:35:27 2009 Subject: [Histonet] Meeting in Virginia May 16th. Message-ID: <37E4BAC017F57141AF64FAA5AEB04CE8033A72A1@wlmmsx01.nemours.org> Hello-o-o-o Histonetter' s. I have had several calls and e-mails from those wishing to attend this Saturday's Seminar's at Old Dominion Higher Education Center, Virginia Beach ....but, can't get their checks in time to get the $10 registration fee waived. If you have an approval..please just fax your approval: cbarone 302-651-5010...you may bring the check with you, or pay yourself at the door with personal check, that is fine. No credit cards, unfortunately...but we would love to see you there. There will be open registration at the door...and you do not need to be any state member to attend. Pre-registration closes tomorrow...but discounts are still available to the state members at the door. Even if you have no state affiliation and need to pay at the door...you will receive 5 CEU's with a full day registration... or you can register at the door for a half day session and earn 2 or 3 CEU's depending on the workshop you choose....for as little as $50. We would especially like to see you Virginians....and have the time to to meet you and help the VA Society to become activated as a state society, whether or not you wish to become a constituent state of NSH or member. It is your society, and you will choose. We offer only the support to help you suceed, as sister states in the region...But, remember, you can always attend just to receive the CEU's...with no strings! Call if you have questions: 302-563-4392! remember there will be a continental breakfast for all and lunch included for full day attendees! Call or e-mail to sign up soon!!!! Meeting info: We couldn't make it any easier or less expensive for you to Learn and Earn those needed CEU's... May 16th "Learn and Earn - Saturday Seminars", hosted by the regional sister states of DE, MD. PA and NJ in support of re-activating the Virginia State Society. 2 excellent workshops, 5 CEU's, 1 continental breakfast and a free lunch will full day registration.... registration is accepted at the door and $5 discount will be given on each seminar to attendees with state memberships in DE, MD, PA and NJ...or. with a new, or renewed, membership in the Virginia Society. You can earn 5 CEU's in a single day with lunch provided. So why are you staying home on May 16th? Register now!!! This is how every state society supports it's sister states....and tech's! Seminars: Antibody Panels in the Anatomic Pathology Lab Multiplex Stains for the Anatomic Pathology Lab For more information on how to register: cbarone@nemours.org PS: Education is professional development! A requirement for ASCP and and investment in yourself durning competitive times!! From eric.halpern <@t> comphealth.com Tue May 12 12:15:48 2009 From: eric.halpern <@t> comphealth.com (eric.halpern@comphealth.com) Date: Tue May 12 12:15:57 2009 Subject: [Histonet] Job Openings Message-ID: Good Afternoon- How do I review the job openings on the Histonet site? Eric Halpern Sr.Search Consultant, Lab Sciences and Imaging Division Permanent Placement Division CompHealth Associates, Inc. 6451 North Federal Highway, Suite 702 Fort Lauderdale, Florida 33308 Phone: 866-782-9029, ext. 5852 Cell 954-336-8913 Fax: 800-420-2329 email: eric.halpern@comphealth.com www.comphealth.com Please note: "This is a commercial email from CompHealth Associates, Inc. If you do not want to receive future emails from CompHealth Associates, Inc. , please reply to the sender of this email and ask to be removed from our list." Customer service is the key to success. Are you satisfied with your experience? Send your comments to my manager at valerie.patterson@comphealth.com. From wilson_c <@t> ricerca.com Tue May 12 14:00:59 2009 From: wilson_c <@t> ricerca.com (Wilson, Carol) Date: Tue May 12 14:01:06 2009 Subject: [Histonet] Lab to do Possible EM as well as preferred fixative Message-ID: <9D443EB9D0270143B5AAF190CB1A58A309746E46@dogwood.ricerca.com> Hi All, We are going to be collecting rat ileum for possible EM after completion of our study. I am looking for a lab to possibly process these specimens as well as what most would consider the preferred choice of fixative and storage while we are holding the specimens until the decision is made to process them. Thanks in advance for replies. Carol Carol Wilson, HT(ASCP) Lead Technician/Histology From athorn <@t> bmhvt.org Tue May 12 14:15:39 2009 From: athorn <@t> bmhvt.org (Anita Thorn) Date: Tue May 12 14:14:05 2009 Subject: [Histonet] Lean Consultants Message-ID: <602863D272B56749A70CBA315D7DC70203B328E2@bmhexch.bmhvt.org> Are there any companies out there that go around helping set up LEAN Histology laboratories? If anybody can supply some company names I'd appreciate it. _______________________________________________________________ The information contained in, or attached to, this e-mail, may contain confidential information and is intended solely for the use of the individual or entity to whom it is addressed and may be subject to legal privilege. If you have received this e-mail in error you should notify the sender immediately by reply e-mail, delete the message from your system and notify your system manager. Please do not copy it for any purpose, or disclose its contents to any other person. The views or opinions presented in this e-mail are solely those of the author and do not necessarily represent those of the company. The recipient should check this e-mail and any attachments for the presence of viruses. The company accepts no liability for any damage caused, directly or indirectly, by any virus transmitted in this email. _______________________________________________________________ From rcharles <@t> state.pa.us Tue May 12 14:35:05 2009 From: rcharles <@t> state.pa.us (Charles, Roger) Date: Tue May 12 14:35:09 2009 Subject: [Histonet] Looking for Jan Shivers Message-ID: <3809C163DC1DA54AA534B3C7794D07B633CEA2FAD9@ENHBGMBX01.PA.LCL> Please contact me off list. I have questions on toxoplasma gondii IHC testing. Thanks Roger Roger Charles Microbiologist Pennsylvania Veterinary Laboratory 2305 N Cameron St Harrisburg, PA 17110 717-787-8808 rcharles@state.pa.us No trees were hurt in the sending of this email, However many electrons were severely inconvenienced! From mikael.niku <@t> helsinki.fi Tue May 12 16:04:14 2009 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Tue May 12 16:04:22 2009 Subject: [Histonet] Basement membrane "counterstain" for IHC? References: <4A080D00.7400.0077.1@harthosp.org> Message-ID: <9A207AD48BD646C2A5EC0E56B2CC6BDC@Lentomato2> Dear Histonetters, I'm doing IHC for intraepithelial lymphocytes using paraffin sections of murine small intestines. To make counting the cells easier and more reliable, I'd like to add a basement membrane stain to the protocol. Is there a simple and compatible histological basement membrane stain you would like to recommend here? For IHC, I'm using a standard biotin and peroxidase based detection system and the DAB substrate (brown signal). The tissues are PFA fixed material, and the antibodies require heat-induced epitope retrieval in microwave oven. The immunostaining is performed either manually (with ThermoShandon coverplates) or using a LabVision autostainer. Normally, I'm using a hematoxylin counterstain (just in water, no eosin) and a water-soluble embedding medium. With best regards, Mikael Niku University of Helsinki, Finland From jcampbell <@t> vdxpathology.com Wed May 13 07:51:33 2009 From: jcampbell <@t> vdxpathology.com (Jennifer Campbell) Date: Wed May 13 07:51:44 2009 Subject: [Histonet] Question about DAB waste Message-ID: <5658CBDB9EAE6545ABE50D2563D81BF821CFC4@VDXSERVER01.vdxpathology.local> We just purchased a Dako Autostainer and are in the process of figuring out how to get rid of the haz waste that we will be generating. The machine separates the haz from non-haz, thankfully. I've been communicating with our waste facility and have been having some difficulty. They want to know a lot of specifics like, how much waste we will be brining to them at a time and how often, what sort of containers we will be bringing it in etc. Do you let the haz waste carboy get to a certain level before emptying it? Is it ok to pour this waste into plastic containers of some sort (we were going to use empty distilled water bottles) to transport them to the facility? I would appreciate any insight. Thank you. Jennifer C. From asmith <@t> mail.barry.edu Wed May 13 07:57:02 2009 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Wed May 13 07:59:08 2009 Subject: [Histonet] RE: Question about DAB waste In-Reply-To: <5658CBDB9EAE6545ABE50D2563D81BF821CFC4@VDXSERVER01.vdxpathology.local> References: <5658CBDB9EAE6545ABE50D2563D81BF821CFC4@VDXSERVER01.vdxpathology.local> Message-ID: We keep our DAB waste in a plastic container and give it to our waste disposal company every 6 months. No one has objected to this procedure. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Podiatric Medicine Miami Shores, FL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jennifer Campbell Sent: Wednesday, May 13, 2009 8:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Question about DAB waste We just purchased a Dako Autostainer and are in the process of figuring out how to get rid of the haz waste that we will be generating. The machine separates the haz from non-haz, thankfully. I've been communicating with our waste facility and have been having some difficulty. They want to know a lot of specifics like, how much waste we will be brining to them at a time and how often, what sort of containers we will be bringing it in etc. Do you let the haz waste carboy get to a certain level before emptying it? Is it ok to pour this waste into plastic containers of some sort (we were going to use empty distilled water bottles) to transport them to the facility? I would appreciate any insight. Thank you. Jennifer C. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akbitting <@t> geisinger.edu Wed May 13 08:08:12 2009 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Wed May 13 08:08:23 2009 Subject: [Histonet] Lean Consultants In-Reply-To: <602863D272B56749A70CBA315D7DC70203B328E2@bmhexch.bmhvt.org> References: <602863D272B56749A70CBA315D7DC70203B328E2@bmhexch.bmhvt.org> Message-ID: <4A0A8DFC.2B7F.00C9.0@geisinger.edu> Here's the website for consulting group we used: www.sprickstegall.com Angie Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 No trees were hurt in the sending of this email However many electrons were severly inconvienienced! >>> "Anita Thorn" 5/12/2009 3:15 PM >>> Are there any companies out there that go around helping set up LEAN Histology laboratories? If anybody can supply some company names I'd appreciate it. _______________________________________________________________ The information contained in, or attached to, this e-mail, may contain confidential information and is intended solely for the use of the individual or entity to whom it is addressed and may be subject to legal privilege. If you have received this e-mail in error you should notify the sender immediately by reply e-mail, delete the message from your system and notify your system manager. Please do not copy it for any purpose, or disclose its contents to any other person. The views or opinions presented in this e-mail are solely those of the author and do not necessarily represent those of the company. The recipient should check this e-mail and any attachments for the presence of viruses. The company accepts no liability for any damage caused, directly or indirectly, by any virus transmitted in this email. _______________________________________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Bitting, Angela TEL;WORK:570-271-6844 ORG:;Histology EMAIL;WORK;PREF;NGW:AKBITTING@geisinger.edu N:Bitting;Angela END:VCARD From cbrya <@t> lexclin.com Wed May 13 08:12:20 2009 From: cbrya <@t> lexclin.com (Carol Bryant) Date: Wed May 13 08:12:32 2009 Subject: [Histonet] RE: Question about DAB waste In-Reply-To: References: <5658CBDB9EAE6545ABE50D2563D81BF821CFC4@VDXSERVER01.vdxpathology.local> Message-ID: <37A1F9CAB9E21C41B39F3653B620D13E093BA254@exchange2003.lc.local> On this subject at our recent CAP inspection a question came up about the labeling of the waste containers that held the DAB waste from our Ventana stainer. We had the container labeled as "Hazardous Chemical Waste". We also keep it in a large plastic carboy which our maintenance dept. then empties into a large metal can that is picked up by a hazardous waste disposal company as the cans become full. The inspector insisted that the waste container itself from our Ventana stainer should have listed every chemical that could possibly be in the waste. How are the DAB waste containers labeled at other facilities? Thank you, Carol -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Smith, Allen Sent: Wednesday, May 13, 2009 8:57 AM To: 'Jennifer Campbell' Cc: 'Histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: Question about DAB waste We keep our DAB waste in a plastic container and give it to our waste disposal company every 6 months. No one has objected to this procedure. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Podiatric Medicine Miami Shores, FL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jennifer Campbell Sent: Wednesday, May 13, 2009 8:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Question about DAB waste We just purchased a Dako Autostainer and are in the process of figuring out how to get rid of the haz waste that we will be generating. The machine separates the haz from non-haz, thankfully. I've been communicating with our waste facility and have been having some difficulty. They want to know a lot of specifics like, how much waste we will be brining to them at a time and how often, what sort of containers we will be bringing it in etc. Do you let the haz waste carboy get to a certain level before emptying it? Is it ok to pour this waste into plastic containers of some sort (we were going to use empty distilled water bottles) to transport them to the facility? I would appreciate any insight. Thank you. Jennifer C. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by the State of Kentucky and/or Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient and have received this communication in error, please contact the sender and destroy all copies of the original message. Thank you. From JWeems <@t> sjha.org Wed May 13 08:17:04 2009 From: JWeems <@t> sjha.org (Weems, Joyce) Date: Wed May 13 08:17:07 2009 Subject: [Histonet] RE: Question about DAB waste In-Reply-To: <37A1F9CAB9E21C41B39F3653B620D13E093BA254@exchange2003.lc.local> References: <5658CBDB9EAE6545ABE50D2563D81BF821CFC4@VDXSERVER01.vdxpathology.local> <37A1F9CAB9E21C41B39F3653B620D13E093BA254@exchange2003.lc.local> Message-ID: <5D64396A0D4A5346BEBC759022AAEAA5505E91@ITSSSXM01V6.one.ads.che.org> We had ours analyzed by our Hazardous Waste Disposal Co. They provide labels for the drums. We have a drum for the waste from the special stains, the DAB waste, and the xylene waste from the recycler - labels all provided by the disposal company. j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Carol Bryant Sent: Wednesday, May 13, 2009 9:12 AM To: Smith, Allen; Jennifer Campbell Cc: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Question about DAB waste On this subject at our recent CAP inspection a question came up about the labeling of the waste containers that held the DAB waste from our Ventana stainer. We had the container labeled as "Hazardous Chemical Waste". We also keep it in a large plastic carboy which our maintenance dept. then empties into a large metal can that is picked up by a hazardous waste disposal company as the cans become full. The inspector insisted that the waste container itself from our Ventana stainer should have listed every chemical that could possibly be in the waste. How are the DAB waste containers labeled at other facilities? Thank you, Carol Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. From akbitting <@t> geisinger.edu Wed May 13 08:25:49 2009 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Wed May 13 08:26:02 2009 Subject: [Histonet] Job Openings In-Reply-To: References: Message-ID: <4A0A921D.2B7F.00C9.0@geisinger.edu> We have a part-time dayshift and a full-time night shift opening for a HT. If you're interested here's our web address. www.geisinger.org or call me directly. Angie Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 No trees were hurt in the sending of this email However many electrons were severly inconvienienced! >>> 5/12/2009 1:15 PM >>> Good Afternoon- How do I review the job openings on the Histonet site? Eric Halpern Sr.Search Consultant, Lab Sciences and Imaging Division Permanent Placement Division CompHealth Associates, Inc. 6451 North Federal Highway, Suite 702 Fort Lauderdale, Florida 33308 Phone: 866-782-9029, ext. 5852 Cell 954-336-8913 Fax: 800-420-2329 email: eric.halpern@comphealth.com www.comphealth.com Please note: "This is a commercial email from CompHealth Associates, Inc. If you do not want to receive future emails from CompHealth Associates, Inc. , please reply to the sender of this email and ask to be removed from our list." Customer service is the key to success. Are you satisfied with your experience? Send your comments to my manager at valerie.patterson@comphealth.com. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Bitting, Angela TEL;WORK:570-271-6844 ORG:;Histology EMAIL;WORK;PREF;NGW:AKBITTING@geisinger.edu N:Bitting;Angela END:VCARD From cheastys <@t> svm.vetmed.wisc.edu Wed May 13 09:17:35 2009 From: cheastys <@t> svm.vetmed.wisc.edu (Sandra Cheasty) Date: Wed May 13 09:17:41 2009 Subject: [Histonet] IHC - PGDFR reactive in Canines Message-ID: Hello, Does anyone have experience using PGDFR in canines and if so, is there a canine species control that is known to react with PGDFR? Thank you, Sandy Sandra Cheasty Histology Supervisor UW-Madison School of Veterinary Medicine 608 263-1680 From njoydobro <@t> aol.com Wed May 13 09:25:23 2009 From: njoydobro <@t> aol.com (njoydobro@aol.com) Date: Wed May 13 09:25:51 2009 Subject: [Histonet] xylene substitues Message-ID: <8CBA1EDC951C9EF-7D0-310@webmail-mh26.sysops.aol.com> just wanting to see what everyone's favorite xylene substitute is for clearing slides before coverslipping?? Gene Cleveland Clinic From gmartin <@t> marshallmedical.org Wed May 13 09:50:11 2009 From: gmartin <@t> marshallmedical.org (Martin, Gary) Date: Wed May 13 09:51:09 2009 Subject: [Histonet] xylene substitutes In-Reply-To: <8CBA1EDC951C9EF-7D0-310@webmail-mh26.sysops.aol.com> References: <8CBA1EDC951C9EF-7D0-310@webmail-mh26.sysops.aol.com> Message-ID: <6ED9D4252F278841A0593D3D788AF24C05439AB8@mailsvr.MARSHMED.local> We use Formula 83 ... we have had no problems in all phases. Gary -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of njoydobro@aol.com Sent: Wednesday, May 13, 2009 7:25 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] xylene substitues just wanting to see what everyone's favorite xylene substitute is for clearing slides before coverslipping?? Gene Cleveland Clinic _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histonet.nospam <@t> vneubert.com Wed May 13 09:58:50 2009 From: histonet.nospam <@t> vneubert.com (V. Neubert) Date: Wed May 13 09:58:54 2009 Subject: [Histonet] xylene substitues In-Reply-To: <8CBA1EDC951C9EF-7D0-310@webmail-mh26.sysops.aol.com> References: <8CBA1EDC951C9EF-7D0-310@webmail-mh26.sysops.aol.com> Message-ID: <4A0AE02A.6050602@vneubert.com> N-butyl acetate aka butyl ethanoate. njoydobro@aol.com wrote: > just wanting to see what everyone's favorite xylene substitute is for clearing slides before coverslipping?? > > Gene > Cleveland Clinic > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From lblazek <@t> digestivespecialists.com Wed May 13 10:01:12 2009 From: lblazek <@t> digestivespecialists.com (Blazek, Linda) Date: Wed May 13 10:00:27 2009 Subject: [Histonet] xylene substitutes In-Reply-To: <6ED9D4252F278841A0593D3D788AF24C05439AB8@mailsvr.MARSHMED.local> References: <8CBA1EDC951C9EF-7D0-310@webmail-mh26.sysops.aol.com> <6ED9D4252F278841A0593D3D788AF24C05439AB8@mailsvr.MARSHMED.local> Message-ID: <5A2BD13465E061429D6455C8D6B40E390885D6AA16@IBMB7Exchange.digestivespecialists.com> I second Gary's comments. We use Formula 83 in all phases also. Linda -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martin, Gary Sent: Wednesday, May 13, 2009 10:50 AM To: njoydobro@aol.com; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] xylene substitutes We use Formula 83 ... we have had no problems in all phases. Gary -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of njoydobro@aol.com Sent: Wednesday, May 13, 2009 7:25 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] xylene substitues just wanting to see what everyone's favorite xylene substitute is for clearing slides before coverslipping?? Gene Cleveland Clinic _______________________________________________ Histonet mailing list Histonet@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 hej01 <@t> health.state.ny.us Wed May 13 10:11:54 2009 From: hej01 <@t> health.state.ny.us (Helen E Johnson) Date: Wed May 13 10:12:01 2009 Subject: [Histonet] blood spots Message-ID: <3254_1242227517_n4DFBv5U007555_OFBF84757F.D1D3AFCE-ON852575B5.0052E714-852575B5.00537C5B@notes.health.state.ny.us> Does anyone have protocols for doing histology and hematology on blood spots? Helen Johnson (hej01@health.state.ny.us) IMPORTANT NOTICE: This e-mail and any attachments may contain confidential or sensitive information which is, or may be, legally privileged or otherwise protected by law from further disclosure. It is intended only for the addressee. If you received this in error or from someone who was not authorized to send it to you, please do not distribute, copy or use it or any attachments. Please notify the sender immediately by reply e-mail and delete this from your system. Thank you for your cooperation. From lpaveli1 <@t> hurleymc.com Wed May 13 10:46:53 2009 From: lpaveli1 <@t> hurleymc.com (Lynette Pavelich) Date: Wed May 13 10:47:06 2009 Subject: [Histonet] xylene substitues Message-ID: <4A0AB32D020000EE0002923C@smtp-gw.hurleymc.com> We use Propar by Anatech. It works great. Lynette >>> 05/13/09 10:25 AM >>> just wanting to see what everyone's favorite xylene substitute is for clearing slides before coverslipping?? Gene Cleveland Clinic _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tifei <@t> foxmail.com Wed May 13 10:53:16 2009 From: tifei <@t> foxmail.com (TF) Date: Wed May 13 10:53:36 2009 Subject: [Histonet] xylene substitues References: <4A0AB32D020000EE0002923C@smtp-gw.hurleymc.com> Message-ID: <200905132353106663880@foxmail.com> dHVsYW5lICEhIQ0KDQoNCjIwMDktMDUtMTMgDQoNCg0KDQpURiANCg0KDQoNCreivP7Iy6O6IEx5 bmV0dGUgUGF2ZWxpY2ggDQq3osvNyrG85KO6IDIwMDktMDUtMTMgIDIzOjQ5OjU5IA0KytW8/sjL o7ogbmpveWRvYnJvQGFvbC5jb207IEhpc3RvbmV0QGxpc3RzLnV0c291dGh3ZXN0ZXJuLmVkdSAN CrOty82juiANCtb3zOKjuiBSZTogW0hpc3RvbmV0XSB4eWxlbmUgc3Vic3RpdHVlcyANCiANCldl IHVzZSBQcm9wYXIgYnkgQW5hdGVjaC4gSXQgd29ya3MgZ3JlYXQuDQpMeW5ldHRlDQo+Pj4gPG5q b3lkb2Jyb0Bhb2wuY29tPiAwNS8xMy8wOSAxMDoyNSBBTSA+Pj4NCmp1c3Qgd2FudGluZyB0byBz ZWUgd2hhdCBldmVyeW9uZSdzIGZhdm9yaXRlIHh5bGVuZSBzdWJzdGl0dXRlIGlzIGZvcg0KY2xl YXJpbmcgc2xpZGVzIGJlZm9yZSBjb3ZlcnNsaXBwaW5nPz8gDQpHZW5lDQpDbGV2ZWxhbmQgQ2xp bmljDQpfX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fXw0KSGlz dG9uZXQgbWFpbGluZyBsaXN0DQpIaXN0b25ldEBsaXN0cy51dHNvdXRod2VzdGVybi5lZHUNCmh0 dHA6Ly9saXN0cy51dHNvdXRod2VzdGVybi5lZHUvbWFpbG1hbi9saXN0aW5mby9oaXN0b25ldA0K X19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX18NCkhpc3RvbmV0 IG1haWxpbmcgbGlzdA0KSGlzdG9uZXRAbGlzdHMudXRzb3V0aHdlc3Rlcm4uZWR1DQpodHRwOi8v bGlzdHMudXRzb3V0aHdlc3Rlcm4uZWR1L21haWxtYW4vbGlzdGluZm8vaGlzdG9uZXQNCg== From tifei <@t> foxmail.com Wed May 13 10:53:35 2009 From: tifei <@t> foxmail.com (TF) Date: Wed May 13 10:53:46 2009 Subject: [Histonet] 0.2M borate acid pH 7.0 Message-ID: <200905132353300422228@foxmail.com> My recipe for 0.2M borate acid is: Sodium Borate acid.10 H2O (molecular weight 381.42, Na4B4O4.10H2O) 19.07 gram H2O 1 Litre Adjust pH = 7.0 with 5M HCl Is this fine? 2009-05-11 TF From njoydobro <@t> aol.com Wed May 13 10:54:55 2009 From: njoydobro <@t> aol.com (njoydobro@aol.com) Date: Wed May 13 10:55:33 2009 Subject: [Histonet] peroxidase block Message-ID: <8CBA1FA4B2E3050-7D0-8D2@webmail-mh26.sysops.aol.com> good morning, ???? Is there anyone out there who has eliminated using Peroxidase Block on either IHC or ICC? Thanks, Gene Cleveland Clinic From lpaveli1 <@t> hurleymc.com Wed May 13 11:10:05 2009 From: lpaveli1 <@t> hurleymc.com (Lynette Pavelich) Date: Wed May 13 11:10:19 2009 Subject: [Histonet] Question about DAB waste Message-ID: <4A0AB89D020000EE00029245@smtp-gw.hurleymc.com> We detoxify our DAB using this method from: HAZARDOUS MATERIALS IN THE HISTOPATHOLOGY LABORATORY, by Dapson & Dapson, fourth edition, pg 184. 1. Prepare the following aqeous stock solutions: a. 0.2M potassium permanganate (3.16% or 31.6g KMnO4/liter) b. 2.0M sulfuric acid (11.2% or 112 mL concentrated acid/liter) 2. Dilute DAB solution of necessary so that its concentration does not exceed 0.9 mg/ml. 3. For each 10ml of DAB solution, add: a. 5ml 0.2 M potassium permanganate (3.16% or 31.6 g/liter) b. 5ml 2.0 M sulfuric acid (196 g/liter, 107 ml/liter or 10.7%) 4. Allow mixture to stand for at least 10 hours. It is now non-mutagenic. 5. Decolorize the mixture with ascorbic acid (add powder until color disappears). This too is an oxidation/reduction reaction. 6. Neutralize the decolorized mixture with sodium bicarbonate (test with pH meter, pH paper or dipsticks). Note: we recommend using magnesium hydroxide/oxide instead of sodium bicarbonate. 7. Discard the solution down the drain provided that local wastewater authorities have given their approval. Hope this helps, Lynette >>> "Jennifer Campbell" 05/13/09 8:51 AM >>> We just purchased a Dako Autostainer and are in the process of figuring out how to get rid of the haz waste that we will be generating. The machine separates the haz from non-haz, thankfully. I've been communicating with our waste facility and have been having some difficulty. They want to know a lot of specifics like, how much waste we will be brining to them at a time and how often, what sort of containers we will be bringing it in etc. Do you let the haz waste carboy get to a certain level before emptying it? Is it ok to pour this waste into plastic containers of some sort (we were going to use empty distilled water bottles) to transport them to the facility? I would appreciate any insight. Thank you. Jennifer C. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From esulkosky <@t> gmail.com Wed May 13 11:26:16 2009 From: esulkosky <@t> gmail.com (Eric Sulkosky) Date: Wed May 13 11:26:20 2009 Subject: [Histonet] Question about DAB Waste Message-ID: <6c3840890905130926x5b9974a7yadfc10c064846f43@mail.gmail.com> Have any of you tried the Dab/Out? It is a system that extracts DAB from liquid waste. The liquid passes through a cartridge and captures the DAB, then you can dispose cartridge as a dry solid waste and pour the rest of the liquid down the drain. It is known to be cost effective and environmentally safe. Just an alternative to think of. Hope this helps Eric From asmith <@t> mail.barry.edu Wed May 13 11:52:23 2009 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Wed May 13 11:54:29 2009 Subject: [Histonet] Question about DAB waste In-Reply-To: <4A0AB89D020000EE00029245@smtp-gw.hurleymc.com> References: <4A0AB89D020000EE00029245@smtp-gw.hurleymc.com> Message-ID: We were told that, in Miami-Dade County, we would have to apply for a license to do this. We gave up on the cumbersome licensing procedure. Allen A. Smith, Ph.D. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lynette Pavelich Sent: Wednesday, May 13, 2009 12:10 PM To: histonet@lists.utsouthwestern.edu; jcampbell@vdxpathology.com Subject: Re: [Histonet] Question about DAB waste We detoxify our DAB using this method from: HAZARDOUS MATERIALS IN THE HISTOPATHOLOGY LABORATORY, by Dapson & Dapson, fourth edition, pg 184. 1. Prepare the following aqeous stock solutions: a. 0.2M potassium permanganate (3.16% or 31.6g KMnO4/liter) b. 2.0M sulfuric acid (11.2% or 112 mL concentrated acid/liter) 2. Dilute DAB solution of necessary so that its concentration does not exceed 0.9 mg/ml. 3. For each 10ml of DAB solution, add: a. 5ml 0.2 M potassium permanganate (3.16% or 31.6 g/liter) b. 5ml 2.0 M sulfuric acid (196 g/liter, 107 ml/liter or 10.7%) 4. Allow mixture to stand for at least 10 hours. It is now non-mutagenic. 5. Decolorize the mixture with ascorbic acid (add powder until color disappears). This too is an oxidation/reduction reaction. 6. Neutralize the decolorized mixture with sodium bicarbonate (test with pH meter, pH paper or dipsticks). Note: we recommend using magnesium hydroxide/oxide instead of sodium bicarbonate. 7. Discard the solution down the drain provided that local wastewater authorities have given their approval. Hope this helps, Lynette >>> "Jennifer Campbell" 05/13/09 8:51 AM >>> We just purchased a Dako Autostainer and are in the process of figuring out how to get rid of the haz waste that we will be generating. The machine separates the haz from non-haz, thankfully. I've been communicating with our waste facility and have been having some difficulty. They want to know a lot of specifics like, how much waste we will be brining to them at a time and how often, what sort of containers we will be bringing it in etc. Do you let the haz waste carboy get to a certain level before emptying it? Is it ok to pour this waste into plastic containers of some sort (we were going to use empty distilled water bottles) to transport them to the facility? I would appreciate any insight. Thank you. Jennifer C. _______________________________________________ Histonet mailing list Histonet@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 annigyg <@t> gmail.com Wed May 13 12:09:28 2009 From: annigyg <@t> gmail.com (Anne van Binsbergen) Date: Wed May 13 12:09:33 2009 Subject: [Histonet] vacant post Message-ID: Hello Histonetters My Immuno lead tech has left and I urgently need a replacement Its a full-time position and requires good IHC experience as well as routine Histo experience - an all-rounder Need someone with at least 7-10 years experience as its a Senior post Anyone interested? Send me your CVs -- Anne van Binsbergen (Hope) Abu Dhabi UAE From jcox90 <@t> yahoo.com Wed May 13 13:03:22 2009 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Wed May 13 13:03:24 2009 Subject: [Histonet] Looking for part-time Histology work in Phoenix AZ Message-ID: <344068.5780.qm@web56803.mail.re3.yahoo.com> Hi All, I am looking?for part-time Histology work in Phoenix AZ. I am available for almost any shift, my current employer is very flexible with my hours. I have been in field for 17 years and know all aspects of Histology. Please respond to this email if you have any questions. Thanks!!! Jill Cox HT (ASCP) ? ? ? ? From anh2006 <@t> med.cornell.edu Wed May 13 14:27:02 2009 From: anh2006 <@t> med.cornell.edu (Andrea Hooper) Date: Wed May 13 14:27:08 2009 Subject: [Histonet] mouse CD11c Message-ID: Is anyone working on CD11c in murine tissues? If so, can we exchange emails? ... I have some questions. Andrea Hooper anh2006@med.cornell.edu -- From anh2006 <@t> med.cornell.edu Wed May 13 14:32:08 2009 From: anh2006 <@t> med.cornell.edu (Andrea Hooper) Date: Wed May 13 14:32:17 2009 Subject: [Histonet] mouse CD115 Message-ID: Anyone working with CD115 in mouse? If so, may I contact you as I have a few questions? Andrea Hooper anh2006 @ med.cornell.edu -- From Montina.VanMeter <@t> pbrc.edu Wed May 13 14:49:11 2009 From: Montina.VanMeter <@t> pbrc.edu (Montina Van Meter) Date: Wed May 13 14:49:17 2009 Subject: [Histonet] 2009 LSH Symposium/Convention-Additional Information Message-ID: <4FE7FB862E90E448AE32388E759220E50140A98D@pbrcas31.pbrc.edu> Fellow Histonetters, The Louisiana Society for Histotechnology is pleased to announce the 26th Annual Symposium/Convention: "Your Histeaux Surplus Package" June 12 & 13, 2009 at the Bourbon Orleans Hotel 717 Orleans St. New Orleans, LA 70117 www.bourbonorleans.com The LSH room rates will be honored as long as rooms are available. For those attendees who may want to visit the beautiful sights and sounds of New Orleans, the Bourbon Orleans Hotel will honor the room rates for three days prior and three days after the meeting. Special on-site parking rates for LSH attendees are available as well as public parking around Jackson Square. For reservations call 1-504-523-2222 or 1-866-513-9744 and mention you are with the LSH group. Do to relocation of many of our members we would ask everyone who would like to receive a brochure/membership form in the mail, email or fax, to please contact Tina Van Meter at 225-603-0953, vanmetmj@pbrc.edu or Dixie Benoit at 337-233-1951, dixiehistochick@bellsouth.net. Walk-ins are always welcome! If you are planning on registering the day of the meeting we would appreciate it if you would send us an email or call Dixie or Tina prior to the meeting which will allow the LSH to make arrangements for workshop handouts and food reservations. Pre-registrants will automatically receive a complimentary buffet lunch each day! Please make additional copies of your registration form for co-workers in your lab or facility that might be interested in attending. The LSH would also like to extend the invitation to our fellow technologists and pathologists from surrounding states. We encourage everyone to attend in order to build our networking potential, earn those valuable CEU's and enjoy the beautiful city of New Orleans. There are many local attractions going on during the week of our symposium such as: The Cajun - Zydeco Festival, Seafood Festival and the Creole Tomato Festival all in the French Quarter/Jackson Square area. Free admission to the festivals. Check out the links below: http://www.neworleansonline.com/news/2009/May/vieuxtodo.html http://ww.neworleansonline.com/neworleans/tours/ 2009 LSH State Meeting Workshops: WS#1 - Am I Really Ready for this CAP Inspection? WS#2 - Mouse to Horse: It's Not Human WS#3 - Breast Cancer and the Standardization of HER2/neu Testing WS#4 - Contemporary Trends in Immunohistochemistry WS#5 - Troubleshooting Routine Special Stains WS#6 - Which Code Do I Use? (CPT Coding) WS#7 - Are You REALLY Ready for the Next Catastrophic Event That Will Affect Your Hospital or City? WS#8 - 2 Crazy Cat Ladies Give Their Opinions of Life, Liberty, and Lab Management We have a variety of topics presented by experienced speakers that promises to benefit everyone. The attendees will have access to several scientific vendor exhibits during the entire symposium. The LSH would love to have y'all come on down to the Bourbon Orleans Hotel in the French Quarter! Hope to see you in June! The LSH Committee Montina J. Van Meter, HT (ASCP) Lab Manager Autonomic Neuroscience Pennington Biomedical Research Center 6400 Perkins Rd. Baton Rouge, LA 70791 225-763-2564 From janet.dertien <@t> ttuhsc.edu Wed May 13 15:20:32 2009 From: janet.dertien <@t> ttuhsc.edu (Dertien, Janet) Date: Wed May 13 15:20:36 2009 Subject: [Histonet] Leica cryostat Message-ID: Hello everyone, We have decided to purchase a refurbished Leica cryostat and are trying to decide which model to choose; does anyone have experience with the 1510, the CM 1800 or the 1800? Any feedback would be appreciated. Thanks! Janet From gmartin <@t> marshallmedical.org Wed May 13 15:40:42 2009 From: gmartin <@t> marshallmedical.org (Martin, Gary) Date: Wed May 13 15:40:45 2009 Subject: [Histonet] Leica cryostat In-Reply-To: References: Message-ID: <6ED9D4252F278841A0593D3D788AF24C05478935@mailsvr.MARSHMED.local> We have an 1850 and like very much, but see if you can find one with the ultra violet light for sterilization. It would save a lot of hassle. Gary -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dertien, Janet Sent: Wednesday, May 13, 2009 1:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Leica cryostat Hello everyone, We have decided to purchase a refurbished Leica cryostat and are trying to decide which model to choose; does anyone have experience with the 1510, the CM 1800 or the 1800? Any feedback would be appreciated. Thanks! Janet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Wed May 13 16:09:10 2009 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Wed May 13 16:07:45 2009 Subject: [Histonet] Toulidine blue stain In-Reply-To: <5658CBDB9EAE6545ABE50D2563D81BF821CEE2@VDXSERVER01.vdxpathology.local> References: <5658CBDB9EAE6545ABE50D2563D81BF821CEE2@VDXSERVER01.vdxpathology.local> Message-ID: <4A0B36F6.4050608@umdnj.edu> I have always liked a very simple method from an older edition of Humanson's book. 1. Bring paraffin sections to water. 2. Stain in 0.1% Toluidine Blue in distilled water, 2-5 minutes 3. Rinse thoroughly in distilled water. 4. Dehydrate in 2 changes of acetone, 5 minutes each. Dehydrating in alcohol will wash all of the stain out. 5. Clear in xylene as usual. Use fresh xylene, not solutions containing carry-over alcohol from other procedures. The second advantage of acetone dehydration is that metachromasia of cartilage, mast cells of some species, etc. is preserved. Geoff Jennifer Campbell wrote: > Hi everyone, > > I'm trying to find a protocol for a toulidine blue stain for paraffin > sections. I've looked in different text books and online and can't seem > to find anything helpful. Does anyone have a good protocol for this? I > need to order the stain rather soon but, first I want to know how much > I'm going to need to run the stain. > > Thanks in advance, > > Jen C. > _______________________________________________ > 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 mcauliff@umdnj.edu ********************************************** From shive003 <@t> umn.edu Wed May 13 16:48:22 2009 From: shive003 <@t> umn.edu (Jan Shivers) Date: Wed May 13 16:48:27 2009 Subject: [Histonet] oxytocin receptor IHC fixation and storage Message-ID: <351524BB1F7340A0B63B06B3B8F25C3D@auxs.umn.edu> I'm sending this for a researcher here. He's got a lot of tissue in storage that's still 'wet' (still in various fixatives and bagged up). The tissues have been in fixative for over 10 years. He wants to know if he can still hang onto the tissue for future IHC use, or has it been in fixative too long for the antigen in question to be retrieved successfully. (I don't need a protocol for oxytocin receptor IHC as of yet; I just need an answer to his storage question - because he's moving his materials from one lab space to another.) I've never done oxytocin receptor IHC, so I don't know how difficult it is to retrieve it on the best of days. Is there anyone out there who might know the answer to this question? Thanks in advance. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu From jclark <@t> pcnm.com Wed May 13 18:11:26 2009 From: jclark <@t> pcnm.com (Joanne Clark) Date: Wed May 13 18:11:30 2009 Subject: [Histonet] RE: Histonet Digest, Vol 66, Issue 15 Message-ID: <0CDA5E1E01301F4880A8A7A8BCBDA39C01024966@mail.pcnm.com> Sprick, Stegall & Associates from Salem OH. Their website is www.sprickstegall.com. I attended a conference where they did a one day workshop on how to set up LEAN and they seemed very experienced. Joanne Clark HT Histology Supervisor Pathology Consultants of New Mexico ------------------------------ Message: 3 Date: Tue, 12 May 2009 15:15:39 -0400 From: "Anita Thorn" Subject: [Histonet] Lean Consultants To: Message-ID: <602863D272B56749A70CBA315D7DC70203B328E2@bmhexch.bmhvt.org> Content-Type: text/plain; charset="us-ascii" Are there any companies out there that go around helping set up LEAN Histology laboratories? If anybody can supply some company names I'd appreciate it. _______________________________________________________________ From annigyg <@t> gmail.com Wed May 13 23:35:49 2009 From: annigyg <@t> gmail.com (Anne van Binsbergen) Date: Wed May 13 23:35:55 2009 Subject: [Histonet] Re: vacant post In-Reply-To: References: Message-ID: Dear ALL My signature (see the very bottom of my email) has my location !! ABU DHABI is the capital of the United Arab Emirates - about 160kms from Dubai I am not permitted to discuss salaries but the current monthly salary of a Senior Medical Technologist in this hospital is around 11 500 AED (thats the currency of the UAE) - which is probably somewhere in the region of $3000 a month Remember that I need current proven IHC skills as well as general all-round Histo lab experience Your qualifications should be at least a 3 year Diploma level with a valid licence to practice in Histology Please send me your CVs asap and I will pass them on to our Recruiment department - include your contact details with an email address cheers Anne 2009/5/13 Anne van Binsbergen > Hello Histonetters > My Immuno tech has left and I urgently need a replacement > Its a full-time position and requires good IHC experience as well as > routine Histo experience - an all-rounder > Need someone with at least 7-10 years experience as its a Senior post > Anyone interested? > Send me your CVs > -- -- Anne van Binsbergen (Hope) Abu Dhabi UAE From kimtournear <@t> yahoo.com Thu May 14 10:01:02 2009 From: kimtournear <@t> yahoo.com (Kim Tournear) Date: Thu May 14 10:01:06 2009 Subject: [Histonet] Leica cryostat Message-ID: <920391.60351.qm@web54204.mail.re2.yahoo.com> We have 2 of the CM1850...no problems....keep up the routine and yearly maintenance and they will out live us all....LOL ~Kim Tournear?~ HT (ASCP), QIHC (ASCP) Histology Supervisor Tucson Medical Center Tucson,? AZ ~Don't?let your life end before it begins~ OU Rocks!!!! --- On Wed, 5/13/09, Dertien, Janet wrote: From: Dertien, Janet Subject: [Histonet] Leica cryostat To: histonet@lists.utsouthwestern.edu Date: Wednesday, May 13, 2009, 1:20 PM Hello everyone, We have decided to purchase a refurbished Leica cryostat and are trying to decide which model to choose; does anyone have experience with the 1510, the CM 1800 or the 1800? Any feedback would be appreciated. Thanks! Janet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From billodonnell <@t> catholichealth.net Thu May 14 10:12:10 2009 From: billodonnell <@t> catholichealth.net (O'Donnell, Bill) Date: Thu May 14 10:12:25 2009 Subject: [Histonet] Phenol-Formalin In-Reply-To: <920391.60351.qm@web54204.mail.re2.yahoo.com> References: <920391.60351.qm@web54204.mail.re2.yahoo.com> Message-ID: Happy Friday! Can anyone send me their formulation for a Penol-Formalin fixative? Thanks. William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 - From billodonnell <@t> catholichealth.net Thu May 14 10:31:52 2009 From: billodonnell <@t> catholichealth.net (O'Donnell, Bill) Date: Thu May 14 10:32:02 2009 Subject: [Histonet] Phenol-Formalin In-Reply-To: References: <920391.60351.qm@web54204.mail.re2.yahoo.com> Message-ID: Oops, my bad, just seems like my Friday! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of O'Donnell, Bill Sent: Thursday, May 14, 2009 10:12 AM To: Histonet Subject: [Histonet] Phenol-Formalin Happy Friday! Can anyone send me their formulation for a Penol-Formalin fixative? Thanks. William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 - _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Thu May 14 11:00:05 2009 From: mucram11 <@t> comcast.net (Pamela Marcum) Date: Thu May 14 11:00:08 2009 Subject: [Histonet] Phenol-Formalin In-Reply-To: Message-ID: <1523111073.205501242316805125.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> Oh if only it were Friday.? Got my hopes up I had lost a day.? Pam Marcum ----- Original Message ----- From: "Bill O'Donnell" To: "Bill O'Donnell" , "Histonet" Sent: Thursday, May 14, 2009 11:31:52 AM GMT -05:00 US/Canada Eastern Subject: RE: [Histonet] Phenol-Formalin Oops, my bad, just seems like my Friday! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of O'Donnell, Bill Sent: Thursday, May 14, 2009 10:12 AM To: Histonet Subject: [Histonet] Phenol-Formalin ? Happy Friday! Can anyone send me their formulation for a Penol-Formalin fixative? Thanks. William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 - _______________________________________________ Histonet mailing list Histonet@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 bhewlett <@t> cogeco.ca Thu May 14 11:17:49 2009 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Thu May 14 11:17:46 2009 Subject: [Histonet] Phenol-Formalin References: <920391.60351.qm@web54204.mail.re2.yahoo.com> Message-ID: <0E04B9C6859443EF9BE3C74F7B17A090@mainbox> Hi Bill, Phenol-formalin fixative consists of the addition of 2% w/v phenol to 4% formaldehyde in phosphate buffer. It has been recommended that sequential fixation in phenol-formalin at pH7.0 followed by phenol-formalin at pH5.5, provides superior results. I am attaching the reference in a separate e-mail offline. I used this fixative extensively in the past with good results. Regards, Bryan ----- Original Message ----- From: "O'Donnell, Bill" To: "Histonet" Sent: Thursday, May 14, 2009 11:12 AM Subject: [Histonet] Phenol-Formalin Happy Friday! Can anyone send me their formulation for a Penol-Formalin fixative? Thanks. William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 - _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pbaldwin <@t> theadvisoryboardprogram.com Thu May 14 11:32:08 2009 From: pbaldwin <@t> theadvisoryboardprogram.com (Peter Baldwin) Date: Thu May 14 11:32:42 2009 Subject: [Histonet] xylene substitues Message-ID: Most of the xylene substitutes are considered hazardous waste by the EPA and local municipalities, and must be treated accordingly. Check the MSDS for flash points below 140 F; OSHA regs classify liquids as "flammable" if flash points are 100 F or below. EPA's regulations for all "solid" hazardous waste (which includes liquids) produced by healthcare facilities and others require that they be handled (including monitoring, storage, and disposal) in accordance with EPA's requirements, and specifically state that hazardous wastes generated in the (healthcare) laboratory "cannot be disposed into drains." If being environmentally and "green chemistry" are important, consider using environmentally and employee safe products in order to reduce the toxic and cost impact on both the environment and personnel, as well as to satisfy EPA's hazardous waste reduction requirements. Peter Peter G. Baldwin Director of Sales, Marketing & Business Development pbaldwin@MicronEnvironmental.com Micron Environmental Industries, Inc. Green Chemistry for LifeSM www.MicronEnvironmental.com 1221 Cameron Street Alexandria, VA 22314 703-548-2776 703-548-7988/Fax From cbarone <@t> NEMOURS.ORG Thu May 14 12:12:08 2009 From: cbarone <@t> NEMOURS.ORG (Barone, Carol ) Date: Thu May 14 12:12:18 2009 Subject: [Histonet] Re: VA Beach - Learn and Earn this Saturday!!!! Message-ID: <37E4BAC017F57141AF64FAA5AEB04CE8033A72AF@wlmmsx01.nemours.org> Virginia Histonetter' s: Spread the word!!!! Walk-ins will be welcome! Learn and earn 5 CEU's in a single day at Old Dominion Higher Education Center, Virginia Beach, VA this Saturday , May 16th. Pre-registrations has closed...but, same-day registrations will be held on site for walk-ins...and state discounts will still apply!. So hey Virginians, you can re-activate your old VA membership or sign on as a new member on Saturday...and still receive the discount for the seminars. or just come to the seminars...no strings attached! Just register at the door. This event has been planned for you and all tech's in the region! Again, 5 NSH CEU's have never been so easy for you to accrue for your ASCP requirements or personal development...we all need to stay ahead of the job market in such "economically trying" times. Come, Learn and Network! Truly, we couldn't make it any easier or less expensive for you to Learn and Earn. One Saturday, two seminars, 5 CEU's. Hosted by your regional sister states of DE, MD. PA and NJ in support of Virginina tech's and technologists. Biocare will sponsor the continental breakfast, and we will add a free lunch with every full day registration. So why are you staying home on May 16th? Come on over for a great day of education. Greak topics, great speakers, great to see old friends, and get CEU's! !Seminars: Antibody Panels in the Anatomic Pathology Lab - speaker Tara Kennedy (3 CEU's) $40 Multiplex Stains for the Anatomic Pathology Lab - speaker Kathy Bowden (2 CEU's) $30 $10 on-site registration...but, a $5 discount on each seminar for attendees with state memberhips...it's a wash!!!! For more information on how to register: cbarone@nemours.org ...or just come on over on Satuday! Where else can you have personal development education come right to your door! From jlhowery <@t> yrmc.org Thu May 14 14:33:55 2009 From: jlhowery <@t> yrmc.org (jeff) Date: Thu May 14 14:34:07 2009 Subject: [Histonet] Pricing Message-ID: <000601c9d4ca$ebd1b040$3394640a@yrmc.org> Can anyone tell me what a private lab would charge for special stains to be done? Also a price for Immuno's Thanks Jeff Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From jstaruk <@t> masshistology.com Thu May 14 15:40:54 2009 From: jstaruk <@t> masshistology.com (jstaruk) Date: Thu May 14 15:41:04 2009 Subject: [Histonet] Pricing In-Reply-To: <000601c9d4ca$ebd1b040$3394640a@yrmc.org> Message-ID: All of our prices are posted on our website. Jim _______________________ James E. Staruk HT(ASCP) www.masshistology.com www.nehorselabs.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of jeff Sent: Thursday, May 14, 2009 3:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pricing Can anyone tell me what a private lab would charge for special stains to be done? Also a price for Immuno's Thanks Jeff 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Marilyn.A.Weiss <@t> kp.org Thu May 14 18:01:20 2009 From: Marilyn.A.Weiss <@t> kp.org (Marilyn.A.Weiss@kp.org) Date: Thu May 14 18:00:16 2009 Subject: [Histonet] marilyn weiss will be out of the office Message-ID: I will be out of the office starting 05/14/2009 and will not return until 05/20/2009. I will respond to your message when I return.In my absence please ask for Mary Campbell or Laurie at 619-528-6801 if this is urgent they can contact me. From neil.macintyre <@t> btinternet.com Fri May 15 03:02:48 2009 From: neil.macintyre <@t> btinternet.com (NEIL MACINTYRE) Date: Fri May 15 03:03:00 2009 Subject: [Histonet] CD25 Message-ID: <149570.81748.qm@web86304.mail.ird.yahoo.com> Hi everyone I have a researcher wishing to look at CD25 in canine tissue. Has anyone any information that could help? Many thanks Neil MacIntyre From tomasz.bonda <@t> gmail.com Fri May 15 05:19:47 2009 From: tomasz.bonda <@t> gmail.com (Tomasz Bonda) Date: Fri May 15 05:19:54 2009 Subject: [Histonet] Embeeding frozen tissue in paraffin Message-ID: <35cd78770905150319vdf0ddcbr790a7dc1b4b88cb8@mail.gmail.com> Hello, is there any good method for embeeding fresh frozen tissue (without any fixation) in paraffin? I would be grateful for ANY suggestions. T. Bonda From louise.renton <@t> gmail.com Fri May 15 06:36:25 2009 From: louise.renton <@t> gmail.com (louise renton) Date: Fri May 15 06:36:29 2009 Subject: [Histonet] Embeeding frozen tissue in paraffin In-Reply-To: <35cd78770905150319vdf0ddcbr790a7dc1b4b88cb8@mail.gmail.com> References: <35cd78770905150319vdf0ddcbr790a7dc1b4b88cb8@mail.gmail.com> Message-ID: why would you want to do this? - the water contained in the fresh tissue will not "mix" with the paraffin wax. There would be little purpose in trying. On 5/15/09, Tomasz Bonda wrote: > > Hello, > is there any good method for embeeding fresh frozen tissue (without any > fixation) in paraffin? > I would be grateful for ANY suggestions. > > T. Bonda > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "There are nights when the wolves are silent and only the moon howls". George Carlin No trees were killed in the sending of this message. However, many electrons were terribly inconvenienced. From april.showers.12345 <@t> gmail.com Fri May 15 09:58:01 2009 From: april.showers.12345 <@t> gmail.com (APRIL SHOWERS) Date: Fri May 15 09:58:05 2009 Subject: [Histonet] Embeeding frozen tissue in paraffin In-Reply-To: <35cd78770905150319vdf0ddcbr790a7dc1b4b88cb8@mail.gmail.com> References: <35cd78770905150319vdf0ddcbr790a7dc1b4b88cb8@mail.gmail.com> Message-ID: > is there any good method for embeeding fresh frozen tissue (without any fixation) in paraffin? Wouldn't the molten paraffin would melt the frozen tissue? LOL.... From april.showers.12345 <@t> gmail.com Fri May 15 09:59:21 2009 From: april.showers.12345 <@t> gmail.com (APRIL SHOWERS) Date: Fri May 15 09:59:25 2009 Subject: [Histonet] CD25 In-Reply-To: <149570.81748.qm@web86304.mail.ird.yahoo.com> References: <149570.81748.qm@web86304.mail.ird.yahoo.com> Message-ID: Google is always a good first step: http://www.google.com/#hl=en&q=CD25+in+canine+tissue&btnG=Google+Search&aq=f&oq=CD25+in+canine+tissue&fp=-dx2DTn7Pl4 On Fri, May 15, 2009 at 4:02 AM, NEIL MACINTYRE < neil.macintyre@btinternet.com> wrote: > Hi everyone > > I have a researcher wishing to look at CD25 in canine tissue. Has anyone > any information that could help? > > Many thanks > > Neil MacIntyre > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From tifei <@t> foxmail.com Fri May 15 10:23:57 2009 From: tifei <@t> foxmail.com (TF) Date: Fri May 15 10:24:15 2009 Subject: [Histonet] 70% alcohol fixation of brain Message-ID: <200905152323518852255@foxmail.com> Hi, i dont want to use PFA for the brain fixation (rat). Now I tried to perfuse the rat with saline, followed with 70% alcohol. Then I do post-fixation at room temperature for 24 hours. I also tried saline perfusion, then I directly put the whole brain into 70% alcohol. Is this fine? Also, for dehydration before cutting frozen sections on a cryostat/microtome, should I use 95% alcohol? I am now using 30% sucrose~ 2009-05-15 TF From mpence <@t> grhs.net Fri May 15 10:44:29 2009 From: mpence <@t> grhs.net (Mike Pence) Date: Fri May 15 10:44:34 2009 Subject: [Histonet] Customary fees Message-ID: <661949901A768E4F9CC16D8AF8F2838C017A3B4B@IS-E2K3.grhs.net> Hi All, I am just wanting to get an idea of how many of you charge law firms or legal gathering sites for making copies of pathology reports and sending slides and blocks. It seems that people are suing for any and everything they can these days. I would also like to know what people are charging for their copying and time with these cases. Please respond off line if you don't want to "publish" your fees. Thanks, Mike Pence AP Supervisor GRMC West Burlington, Iowa From mcauliff <@t> umdnj.edu Fri May 15 10:56:40 2009 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri May 15 10:55:16 2009 Subject: [Histonet] 70% alcohol fixation of brain In-Reply-To: <200905152323518852255@foxmail.com> References: <200905152323518852255@foxmail.com> Message-ID: <4A0D90B8.7060002@umdnj.edu> Greetings TF: Good luck freezing a brain that is full of alcohol! Have you checked the freezing point of alcohol? Why are you doing this? Immersing a whole brain in 70% alcohol, why?? 30% sucrose is a cryoprotectant so the brain is not full of holes from ice crystals.Alcohol defeats this purpose. Geoff TF wrote: > Hi, i dont want to use PFA for the brain fixation (rat). > Now I tried to perfuse the rat with saline, followed with 70% alcohol. Then I do post-fixation at room temperature for 24 hours. > I also tried saline perfusion, then I directly put the whole brain into 70% alcohol. > Is this fine? > > Also, for dehydration before cutting frozen sections on a cryostat/microtome, should I use 95% alcohol? I am now using 30% sucrose~ > > 2009-05-15 > > > > TF > _______________________________________________ > 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 mcauliff@umdnj.edu ********************************************** From april.showers.12345 <@t> gmail.com Fri May 15 10:58:32 2009 From: april.showers.12345 <@t> gmail.com (APRIL SHOWERS) Date: Fri May 15 10:58:37 2009 Subject: [Histonet] Customary fees In-Reply-To: <661949901A768E4F9CC16D8AF8F2838C017A3B4B@IS-E2K3.grhs.net> References: <661949901A768E4F9CC16D8AF8F2838C017A3B4B@IS-E2K3.grhs.net> Message-ID: > It seems that people are suing for any and everything they can these days I would too if I was misdiagnosed or otherwise the victim of malpractice involving incorrect/erroneous histopathology. Don't disrespect such victims by lumping them in the "damn litigious people these days!" crowd. On Fri, May 15, 2009 at 11:44 AM, Mike Pence wrote: > Hi All, > > I am just wanting to get an idea of how many of you charge law firms or > legal gathering sites for making copies of pathology reports and sending > slides and blocks. It seems that people are suing for any and everything > they can these days. I would also like to know what people are charging > for their copying and time with these cases. Please respond off line if > you don't want to "publish" your fees. > > Thanks, > Mike Pence > AP Supervisor > GRMC > West Burlington, Iowa > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From april.showers.12345 <@t> gmail.com Fri May 15 11:03:44 2009 From: april.showers.12345 <@t> gmail.com (APRIL SHOWERS) Date: Fri May 15 11:03:47 2009 Subject: [Histonet] 70% alcohol fixation of brain In-Reply-To: <4A0D90B8.7060002@umdnj.edu> References: <200905152323518852255@foxmail.com> <4A0D90B8.7060002@umdnj.edu> Message-ID: > Good luck freezing a brain that is full of alcohol! Have you checked the freezing point of alcohol? > Why are you doing this? I think that moire often than not, people ask before they think ;-) On Fri, May 15, 2009 at 11:56 AM, Geoff McAuliffe wrote: > Greetings TF: > > Good luck freezing a brain that is full of alcohol! Have you checked the > freezing point of alcohol? > Why are you doing this? > Immersing a whole brain in 70% alcohol, why?? > 30% sucrose is a cryoprotectant so the brain is not full of holes from ice > crystals.Alcohol defeats this purpose. > > Geoff > > > > TF wrote: > >> Hi, i dont want to use PFA for the brain fixation (rat). >> Now I tried to perfuse the rat with saline, followed with 70% alcohol. >> Then I do post-fixation at room temperature for 24 hours. >> I also tried saline perfusion, then I directly put the whole brain into >> 70% alcohol. >> Is this fine? >> >> Also, for dehydration before cutting frozen sections on a >> cryostat/microtome, should I use 95% alcohol? I am now using 30% sucrose~ >> >> 2009-05-15 >> >> >> TF _______________________________________________ >> 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 mcauliff@umdnj.edu > ********************************************** > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From kenneth.metzger <@t> aruplab.com Fri May 15 11:12:47 2009 From: kenneth.metzger <@t> aruplab.com (Metzger, Kenneth) Date: Fri May 15 11:12:55 2009 Subject: [Histonet] KP Marker Plus Message-ID: Does anyone know who sells the KP Marker PLUS? I can't seem to find them. Thanks for any help. Ken Kenneth G Metzger HTL(ASCP) ARUP Labs Histology Supervisor 500 Chipeta Way Salt Lake City, Utah 84108-1221 kenneth.metzger@aruplab.com (801) 583-2787 x 3101 - ------------------------------------------------------------------ The information transmitted by this e-mail and any included attachments are from ARUP Laboratories and are intended only for the recipient. The information contained in this message is confidential and may constitute inside or non-public information under international, federal, or state securities laws, or protected health information and is intended only for the use of the recipient. Unauthorized forwarding, printing, copying, distributing, or use of such information is strictly prohibited and may be unlawful. If you are not the intended recipient, please promptly delete this e-mail and notify the sender of the delivery error or you may call ARUP Laboratories Compliance Hot Line in Salt Lake City, Utah USA at (+1 (800) 522-2787 ext. 2100 From BMolinari <@t> heart.thi.tmc.edu Fri May 15 11:16:16 2009 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri May 15 11:16:23 2009 Subject: [Histonet] KP Marker Plus In-Reply-To: References: Message-ID: Mercedes Medical 800-331-2716 Betsy Molinari HT (ASCP) Texas heart Institute Cardiovascular Pathology Houston,TX 77090 832-355-6524 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Metzger, Kenneth Sent: Friday, May 15, 2009 11:13 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] KP Marker Plus Does anyone know who sells the KP Marker PLUS? I can't seem to find them. Thanks for any help. Ken Kenneth G Metzger HTL(ASCP) ARUP Labs Histology Supervisor 500 Chipeta Way Salt Lake City, Utah 84108-1221 kenneth.metzger@aruplab.com (801) 583-2787 x 3101 - ------------------------------------------------------------------ The information transmitted by this e-mail and any included attachments are from ARUP Laboratories and are intended only for the recipient. The information contained in this message is confidential and may constitute inside or non-public information under international, federal, or state securities laws, or protected health information and is intended only for the use of the recipient. Unauthorized forwarding, printing, copying, distributing, or use of such information is strictly prohibited and may be unlawful. If you are not the intended recipient, please promptly delete this e-mail and notify the sender of the delivery error or you may call ARUP Laboratories Compliance Hot Line in Salt Lake City, Utah USA at (+1 (800) 522-2787 ext. 2100 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Fri May 15 11:41:04 2009 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri May 15 11:41:21 2009 Subject: [Histonet] Embeeding frozen tissue in paraffin In-Reply-To: <35cd78770905150319vdf0ddcbr790a7dc1b4b88cb8@mail.gmail.com> Message-ID: <4EBFF65383B74D49995298C4976D1D5E03835CC3@LSRIEXCH1.lsmaster.lifespan.org> I assume you mean the tissue was frozen without fixation, not that you want to put it into paraffin without fixation? No tissue will withstand paraffin processing without fixation. The best way to turn a frozen tissue into a paraffin embedded tissue is to drop the frozen tissue (either plain or in embedding medium, whichever is the case) into formalin while still frozen. Let it thaw and fix in the formalin, using a typical fixation time for a tissue that size, then just process and embed it as you would any tissue. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tomasz Bonda > Sent: Friday, May 15, 2009 6:19 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Embeeding frozen tissue in paraffin > > Hello, > is there any good method for embeeding fresh frozen tissue (without any > fixation) in paraffin? > I would be grateful for ANY suggestions. > > T. Bonda > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From djohnson <@t> mercedesmedical.com Fri May 15 12:09:06 2009 From: djohnson <@t> mercedesmedical.com (Dave Johnson) Date: Fri May 15 12:09:12 2009 Subject: FW: [Histonet] KP Marker Plus Message-ID: Ken I am sorry that our contact info isnt on the pens themselves. We are the U.S. source for the Klinipath KP Pen. Thanks Dave Johnson Mercedes Medical 7590 Commerce Court Sarasota, FL 34243 800-331-2716 www.mercedesmedical.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Metzger, Kenneth Sent: Friday, May 15, 2009 12:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] KP Marker Plus Does anyone know who sells the KP Marker PLUS? I can't seem to find them. Thanks for any help. Ken Kenneth G Metzger HTL(ASCP) ARUP Labs Histology Supervisor 500 Chipeta Way Salt Lake City, Utah 84108-1221 kenneth.metzger@aruplab.com (801) 583-2787 x 3101 - ------------------------------------------------------------------ The information transmitted by this e-mail and any included attachments are from ARUP Laboratories and are intended only for the recipient. The information contained in this message is confidential and may constitute inside or non-public information under international, federal, or state securities laws, or protected health information and is intended only for the use of the recipient. Unauthorized forwarding, printing, copying, distributing, or use of such information is strictly prohibited and may be unlawful. If you are not the intended recipient, please promptly delete this e-mail and notify the sender of the delivery error or you may call ARUP Laboratories Compliance Hot Line in Salt Lake City, Utah USA at (+1 (800) 522-2787 ext. 2100 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From leiker <@t> buffalo.edu Fri May 15 12:17:39 2009 From: leiker <@t> buffalo.edu (Merced M Leiker) Date: Fri May 15 12:17:47 2009 Subject: [Histonet] KP Marker Plus In-Reply-To: References: Message-ID: <5B8662F7AB73B2900665DFD2@CDYwxp1931.ad.med.buffalo.edu> Indeed! My box of 12 pens just arrived today. Cat# KLI KPPEN. www.mercedesmedical.com Merced --On Friday, May 15, 2009 11:16 AM -0500 "Molinari, Betsy" wrote: > Mercedes Medical 800-331-2716 > > Betsy Molinari HT (ASCP) > Texas heart Institute > Cardiovascular Pathology > Houston,TX 77090 > 832-355-6524 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Metzger, > Kenneth > Sent: Friday, May 15, 2009 11:13 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] KP Marker Plus > > Does anyone know who sells the KP Marker PLUS? I can't seem to find > them. Thanks for any help. > > > > Ken > > > > Kenneth G Metzger HTL(ASCP) > > ARUP Labs > > Histology Supervisor > > 500 Chipeta Way > > Salt Lake City, Utah 84108-1221 > > kenneth.metzger@aruplab.com > > (801) 583-2787 x 3101 > > > > > - ------------------------------------------------------------------ > The information transmitted by this e-mail and any included > attachments are from ARUP Laboratories and are intended only for the > recipient. The information contained in this message is confidential > and may constitute inside or non-public information under > international, federal, or state securities laws, or protected health > information and is intended only for the use of the recipient. > Unauthorized forwarding, printing, copying, distributing, or use of > such information is strictly prohibited and may be unlawful. If you > are not the intended recipient, please promptly delete this e-mail > and notify the sender of the delivery error or you may call ARUP > Laboratories Compliance Hot Line in Salt Lake City, Utah USA at (+1 > (800) 522-2787 ext. 2100 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 leiker@buffalo.edu 716-829-6118 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. From leiker <@t> buffalo.edu Fri May 15 12:22:45 2009 From: leiker <@t> buffalo.edu (Merced M Leiker) Date: Fri May 15 12:22:50 2009 Subject: [Histonet] KP Marker Plus In-Reply-To: <5B8662F7AB73B2900665DFD2@CDYwxp1931.ad.med.buffalo.edu> References: <5B8662F7AB73B2900665DFD2@CDYwxp1931.ad.med.buffalo.edu> Message-ID: By the way, they are only $55/box... :-) --On Friday, May 15, 2009 1:17 PM -0400 Merced M Leiker wrote: > Indeed! My box of 12 pens just arrived today. Cat# KLI KPPEN. > www.mercedesmedical.com > > Merced > > --On Friday, May 15, 2009 11:16 AM -0500 "Molinari, Betsy" > wrote: > >> Mercedes Medical 800-331-2716 >> >> Betsy Molinari HT (ASCP) >> Texas heart Institute >> Cardiovascular Pathology >> Houston,TX 77090 >> 832-355-6524 >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Metzger, >> Kenneth >> Sent: Friday, May 15, 2009 11:13 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] KP Marker Plus >> >> Does anyone know who sells the KP Marker PLUS? I can't seem to find >> them. Thanks for any help. >> >> >> >> Ken >> >> >> >> Kenneth G Metzger HTL(ASCP) >> >> ARUP Labs >> >> Histology Supervisor >> >> 500 Chipeta Way >> >> Salt Lake City, Utah 84108-1221 >> >> kenneth.metzger@aruplab.com >> >> (801) 583-2787 x 3101 >> >> >> >> >> - ------------------------------------------------------------------ >> The information transmitted by this e-mail and any included >> attachments are from ARUP Laboratories and are intended only for the >> recipient. The information contained in this message is confidential >> and may constitute inside or non-public information under >> international, federal, or state securities laws, or protected health >> information and is intended only for the use of the recipient. >> Unauthorized forwarding, printing, copying, distributing, or use of >> such information is strictly prohibited and may be unlawful. If you >> are not the intended recipient, please promptly delete this e-mail >> and notify the sender of the delivery error or you may call ARUP >> Laboratories Compliance Hot Line in Salt Lake City, Utah USA at (+1 >> (800) 522-2787 ext. 2100 >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > Merced M Leiker > Cardiovascular Medicine > 348 Biomedical Research Building > State University of New York at Buffalo > 3435 Main St, Buffalo, NY 14214 > leiker@buffalo.edu > 716-829-6118 > > No trees were harmed in the sending of this email. > However, many electrons were severely inconvenienced. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 leiker@buffalo.edu 716-829-6118 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. From april.showers.12345 <@t> gmail.com Fri May 15 12:38:08 2009 From: april.showers.12345 <@t> gmail.com (APRIL SHOWERS) Date: Fri May 15 12:38:12 2009 Subject: [Histonet] KP Marker Plus In-Reply-To: References: Message-ID: > Does anyone know who sells the KP Marker PLUS? I can't seem to find them. Thanks for any help. Did you try Googling it? http://www.google.com/#hl=en&q=KP+Marker+Plus&btnG=Google+Search&aq=f&oq=KP+Marker+Plus&fp=-dx2DTn7Pl4 On Fri, May 15, 2009 at 12:12 PM, Metzger, Kenneth < kenneth.metzger@aruplab.com> wrote: > Does anyone know who sells the KP Marker PLUS? I can't seem to find > them. Thanks for any help. > > > > Ken > > > > Kenneth G Metzger HTL(ASCP) > > ARUP Labs > > Histology Supervisor > > 500 Chipeta Way > > Salt Lake City, Utah 84108-1221 > > kenneth.metzger@aruplab.com > > (801) 583-2787 x 3101 > > > > > - ------------------------------------------------------------------ > The information transmitted by this e-mail and any included > attachments are from ARUP Laboratories and are intended only for the > recipient. The information contained in this message is confidential > and may constitute inside or non-public information under > international, federal, or state securities laws, or protected health > information and is intended only for the use of the recipient. > Unauthorized forwarding, printing, copying, distributing, or use of > such information is strictly prohibited and may be unlawful. If you > are not the intended recipient, please promptly delete this e-mail > and notify the sender of the delivery error or you may call ARUP > Laboratories Compliance Hot Line in Salt Lake City, Utah USA at (+1 > (800) 522-2787 ext. 2100 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From pkarlisch <@t> hmc.psu.edu Fri May 15 13:01:25 2009 From: pkarlisch <@t> hmc.psu.edu (Patricia Karlisch) Date: Fri May 15 13:01:46 2009 Subject: [Histonet] Embedding Frozen Tissue in Paraffin/ Message-ID: <4A0D75B5.07B7.008C.0@hmc.psu.edu> I agree with Paul Monfils, there needs to be fixation prior to embedding in paraffin plus the tissue needs to be infiltrated or the paraffin will not penetrate the frozen tissue. Even if you are not looking for infiltration, the hot wax will eventually harden and the wet tissue will fall out or shrink. Over long term storage the tissue will decay or grow lots of fungus and mold. Pat Karlisch Supervisor, Histology, Pathology and Laboratory Medicine Penn State Milton S. Hershey Medical Center Mail Code H179 Hershey, PA 17033 Phone (717) 531-6072 Fax: (717) 531- 7741 email: pkarlisch@psu.edu *****E-Mail Confidentiality Notice***** This message (including any attachments) contains information intended for a specific individual(s) and purpose that may be privileged, confidential or otherwise protected from disclosure pursuant to applicable law. Any inappropriate use, distribution or copying of the message is strictly prohibited and may subject you to criminal or civil penalty. If you have received this transmission in error, please reply to the sender indicating this error and delete the transmission from your system immediately. From tifei <@t> foxmail.com Fri May 15 13:14:56 2009 From: tifei <@t> foxmail.com (=?utf-8?B?VEY=?=) Date: Fri May 15 13:15:20 2009 Subject: =?utf-8?B?UmU6IFJlOiBbSGlzdG9uZXRdIDcwJSBhbGNvaG9sIGZpeGF0aW9uIG9mIGJyYWlu?= References: <200905152323518852255@foxmail.com>, <4A0D90B8.7060002@umdnj.edu> Message-ID: <200905160214514154292@foxmail.com> thanks to the reply. some antigens will be damaged by PFA fixation and can not be retreieval. the shock frozen section with acetone is of quite bad tissue quality. therefore I am trying alcohol! 2009-05-16 TF ???? Geoff McAuliffe ????? 2009-05-15 23:55:15 ???? tifei ??? Histonet@lists.utsouthwestern.edu ??? Re: [Histonet] 70% alcohol fixation of brain Greetings TF: Good luck freezing a brain that is full of alcohol! Have you checked the freezing point of alcohol? Why are you doing this? Immersing a whole brain in 70% alcohol, why?? 30% sucrose is a cryoprotectant so the brain is not full of holes from ice crystals.Alcohol defeats this purpose. Geoff TF wrote: > Hi, i dont want to use PFA for the brain fixation (rat). > Now I tried to perfuse the rat with saline, followed with 70% alcohol. Then I do post-fixation at room temperature for 24 hours. > I also tried saline perfusion, then I directly put the whole brain into 70% alcohol. > Is this fine? > > Also, for dehydration before cutting frozen sections on a cryostat/microtome, should I use 95% alcohol? I am now using 30% sucrose~ > > 2009-05-15 > > > > TF > _______________________________________________ > 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 mcauliff@umdnj.edu ********************************************** From carl.hobbs <@t> kcl.ac.uk Fri May 15 13:40:33 2009 From: carl.hobbs <@t> kcl.ac.uk (Hobbs, Carl) Date: Fri May 15 13:40:57 2009 Subject: [Histonet] Re: Embeeding frozen tissue in paraffin Message-ID: <11D9615B89C10747B1C985966A63D7CA2961FE0E0C@KCL-MAIL04.kclad.ds.kcl.ac.uk> Embedding frozen tissue in Pwax is very possible, Tomasz. If your tissue has been frozen NOT using OCT, just place the frozen tissue into acetone that has been cooled to -20C. x3 changes of -20C acetone , then immediately place tissue into molten pwax x3 changes. Agitate frequently, gently. I assume that your specimens are small: you will have more problems with morphology if you have BIG pieces of tissue. I have done this with whole mouse cerebellum and the pwax H&Es looked OK...I never went further due to a job change. (I used x3 acetone changes over 24Hrs, then x3 Pwax over a period of 3Hrs) Very good Q...please keep experimenting and asking. However, it is always best to give a reason for your Q, then you might not get questions to your questions ;-) So, why do you want to do this? Carl. From kris_dee <@t> verizon.net Fri May 15 14:30:15 2009 From: kris_dee <@t> verizon.net (Krista Jackson) Date: Fri May 15 14:30:19 2009 Subject: [Histonet] HT training in Virginia Message-ID: <596916.17416.qm@web84411.mail.ac2.yahoo.com> I am a MLT registry eligible technician. I received my MLT training in the military. I would like to get into histology. I am looking for an opportunity to gain experience, either through working or and internship. I have no prior histology training but I am eager to learn and I am a fast learner. I live in the Richmond area and I am willing to travel no more than 2 hours. If given an opportunity to work, this training would be my main priority. Anyway, if anyone can help me out it would be greatly appreciated. Thanks! Krista kris_dee@verizon.net From arvidsonkristen <@t> yahoo.com Fri May 15 15:04:41 2009 From: arvidsonkristen <@t> yahoo.com (kristen arvidson) Date: Fri May 15 15:04:44 2009 Subject: [Histonet] QA Manual Question Message-ID: <174282.82780.qm@web65712.mail.ac4.yahoo.com> Hi All...Again.? I guess I have a lot of questions lately.? I have asked this before but I would like to ask it again.? What are people doing for QA/QC?? I have been asked by management to write a QA manual.? I have some ideas but not really sure where to start.? Thanks in advance for any input. From tifei <@t> foxmail.com Sat May 16 11:48:19 2009 From: tifei <@t> foxmail.com (TF) Date: Sat May 16 11:48:37 2009 Subject: [Histonet] Chemicals that inactivate the primary antibody Message-ID: <200905170048141643286@foxmail.com> Hi all, just wonder what kind of treatments/chemicals can complete block the binding of 2nd antibody to binded primary antibody on antigen? I tried HCl , but does not damage all the primaryantibody binding sites - i can still see staining pattern finally. 2009-05-17 TF From rjbuesa <@t> yahoo.com Sat May 16 11:54:20 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat May 16 11:54:24 2009 Subject: [Histonet] Chemicals that inactivate the primary antibody Message-ID: <895404.47773.qm@web65707.mail.ac4.yahoo.com> Try bleach and you will destroy everything! Ren? J. --- On Sat, 5/16/09, TF wrote: From: TF Subject: [Histonet] Chemicals that inactivate the primary antibody To: "Histonet@lists.utsouthwestern.edu" Date: Saturday, May 16, 2009, 12:48 PM Hi all, just wonder what kind of treatments/chemicals can complete block the binding of 2nd antibody to binded primary antibody on antigen? I tried HCl , but does not damage all the primaryantibody binding sites - i can still see staining pattern finally. 2009-05-17 TF _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Sat May 16 12:19:02 2009 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Sat May 16 12:19:09 2009 Subject: [Histonet] 70% alcohol fixation of brain In-Reply-To: <200905160214514154292@foxmail.com> References: <200905152323518852255@foxmail.com>, <4A0D90B8.7060002@umdnj.edu> <200905160214514154292@foxmail.com> Message-ID: U would be better off freezing the brain unfixed than trying to fix in alcohol and freeze it, alcohol fixation is not recommended for freezing tissue. Just freeze slices of the brain or a whole rat or mouse brain and then make sections and fix the sections in alcohol/acetone mixture. Sectioning is not easy but this is a better approach than fixing in alcohol and then trying to freeze. I would try to get some frozen sections from the unfixed frozen brain for use with antibodies not formalin friendly and then thaw fix the frozen block in formalin for several day, infiltrate it in 30% sucrose and refreeze, you should be able to get better sections then, but the tissue has to be fixed before you can cryoprotect in sucrose. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 email pruegg@ihctech.net website www.ihctech.net IHC Resource Group www.ihcrg.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of TF Sent: Friday, May 15, 2009 12:15 PM To: Geoff McAuliffe Cc: Histonet@lists.utsouthwestern.edu Subject: Re: Re: [Histonet] 70% alcohol fixation of brain thanks to the reply. some antigens will be damaged by PFA fixation and can not be retreieval. the shock frozen section with acetone is of quite bad tissue quality. therefore I am trying alcohol! 2009-05-16 TF ???? Geoff McAuliffe ????? 2009-05-15 23:55:15 ???? tifei ??? Histonet@lists.utsouthwestern.edu ??? Re: [Histonet] 70% alcohol fixation of brain Greetings TF: Good luck freezing a brain that is full of alcohol! Have you checked the freezing point of alcohol? Why are you doing this? Immersing a whole brain in 70% alcohol, why?? 30% sucrose is a cryoprotectant so the brain is not full of holes from ice crystals.Alcohol defeats this purpose. Geoff TF wrote: > Hi, i dont want to use PFA for the brain fixation (rat). > Now I tried to perfuse the rat with saline, followed with 70% alcohol. Then I do post-fixation at room temperature for 24 hours. > I also tried saline perfusion, then I directly put the whole brain into 70% alcohol. > Is this fine? > > Also, for dehydration before cutting frozen sections on a cryostat/microtome, should I use 95% alcohol? I am now using 30% sucrose~ > > 2009-05-15 > > > > TF > _______________________________________________ > 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 mcauliff@umdnj.edu ********************************************** From pruegg <@t> ihctech.net Sat May 16 12:21:56 2009 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Sat May 16 12:22:00 2009 Subject: [Histonet] Embeeding frozen tissue in paraffin In-Reply-To: <4EBFF65383B74D49995298C4976D1D5E03835CC3@LSRIEXCH1.lsmaster.lifespan.org> References: <35cd78770905150319vdf0ddcbr790a7dc1b4b88cb8@mail.gmail.com> <4EBFF65383B74D49995298C4976D1D5E03835CC3@LSRIEXCH1.lsmaster.lifespan.org> Message-ID: <89DA385F0C0C498099333D1225112462@ihctechq9h2qof> No you need to thaw/fix in formalin or alcoholic fixative in order to process and embed in paraffin. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 email pruegg@ihctech.net website www.ihctech.net IHC Resource Group www.ihcrg.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Friday, May 15, 2009 10:41 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Embeeding frozen tissue in paraffin I assume you mean the tissue was frozen without fixation, not that you want to put it into paraffin without fixation? No tissue will withstand paraffin processing without fixation. The best way to turn a frozen tissue into a paraffin embedded tissue is to drop the frozen tissue (either plain or in embedding medium, whichever is the case) into formalin while still frozen. Let it thaw and fix in the formalin, using a typical fixation time for a tissue that size, then just process and embed it as you would any tissue. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tomasz Bonda > Sent: Friday, May 15, 2009 6:19 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Embeeding frozen tissue in paraffin > > Hello, > is there any good method for embeeding fresh frozen tissue (without any > fixation) in paraffin? > I would be grateful for ANY suggestions. > > T. Bonda > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Sat May 16 12:25:04 2009 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Sat May 16 12:25:09 2009 Subject: [Histonet] CD25 In-Reply-To: References: <149570.81748.qm@web86304.mail.ird.yahoo.com> Message-ID: <1BDE2FB8ECA64FFBB66E44505B82A6CC@ihctechq9h2qof> The NSH IHC Resource Group is also a good source for info on IHC, NSH members can join the group online at www.ihcrg.org Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 email pruegg@ihctech.net website www.ihctech.net IHC Resource Group www.ihcrg.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of APRIL SHOWERS Sent: Friday, May 15, 2009 8:59 AM To: NEIL MACINTYRE Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] CD25 Google is always a good first step: http://www.google.com/#hl=en&q=CD25+in+canine+tissue&btnG=Google+Search&aq=f &oq=CD25+in+canine+tissue&fp=-dx2DTn7Pl4 On Fri, May 15, 2009 at 4:02 AM, NEIL MACINTYRE < neil.macintyre@btinternet.com> wrote: > Hi everyone > > I have a researcher wishing to look at CD25 in canine tissue. Has anyone > any information that could help? > > Many thanks > > Neil MacIntyre > _______________________________________________ > Histonet mailing list > Histonet@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 tifei <@t> foxmail.com Sat May 16 12:37:58 2009 From: tifei <@t> foxmail.com (=?utf-8?B?VEY=?=) Date: Sat May 16 12:38:07 2009 Subject: =?utf-8?B?UmU6IFJlOiBbSGlzdG9uZXRdIENoZW1pY2FscyB0aGF0IGluYWN0aXZhdGUgdGhlIHByaW1hcnkgYW50aWJvZHk=?= References: <895404.47773.qm@web65707.mail.ac4.yahoo.com> Message-ID: <200905170137532225288@foxmail.com> Hi, but I dont want to destory everything.... I am trying to see if I can prevent the binding of another 2nd antibody to the primary antibody for the first antigen! There fore you can perform double staining from same sepecies-derived primary antibodies! 2009-05-17 TF ???? Rene J Buesa ????? 2009-05-17 00:54:26 ???? Histonet@lists.utsouthwestern.edu; tifei ??? ??? Re: [Histonet] Chemicals that inactivate the primary antibody Try bleach and you will destroy everything! Ren? J. --- On Sat, 5/16/09, TF wrote: From: TF Subject: [Histonet] Chemicals that inactivate the primary antibody To: "Histonet@lists.utsouthwestern.edu" Date: Saturday, May 16, 2009, 12:48 PM Hi all, just wonder what kind of treatments/chemicals can complete block the binding of 2nd antibody to binded primary antibody on antigen? I tried HCl , but does not damage all the primaryantibody binding sites - i can still see staining pattern finally. 2009-05-17 TF _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tifei <@t> foxmail.com Sat May 16 12:42:16 2009 From: tifei <@t> foxmail.com (TF) Date: Sat May 16 12:42:29 2009 Subject: =?utf-8?B?UmU6IFJFOiBSZTogW0hpc3RvbmV0XSA3MCUgYWxjb2hvbCBmaXhhdGlvbiBvZiBicmFpbg==?= References: <200905152323518852255@foxmail.com>, <4A0D90B8.7060002@umdnj.edu>, <200905160214514154292@foxmail.com>, Message-ID: <200905170142114499354@foxmail.com> I did try the frozen section from fresh tissue...and fix them in cold acetone for 10 min, transfer to cold PBS for 30 min, then wash in PBS at RT, begin IHC. But the tissue quality is really tooooooooo bad. You always saw pieces of tissues floating, fall in to small pieces and gone when you washing the slide... Under the microscope...the background is high, and you can only see granule-like existences filled by small cavities. Actually I frozed the OCT-embedded tissue with liquid nitrogen..dont know why the tissue quality is still so bad! 2009-05-17 TF ???? Patsy Ruegg ????? 2009-05-17 01:19:09 ???? tifei@foxmail.com; 'Geoff McAuliffe' ??? Histonet@lists.utsouthwestern.edu ??? RE: Re: [Histonet] 70% alcohol fixation of brain U would be better off freezing the brain unfixed than trying to fix in alcohol and freeze it, alcohol fixation is not recommended for freezing tissue. Just freeze slices of the brain or a whole rat or mouse brain and then make sections and fix the sections in alcohol/acetone mixture. Sectioning is not easy but this is a better approach than fixing in alcohol and then trying to freeze. I would try to get some frozen sections from the unfixed frozen brain for use with antibodies not formalin friendly and then thaw fix the frozen block in formalin for several day, infiltrate it in 30% sucrose and refreeze, you should be able to get better sections then, but the tissue has to be fixed before you can cryoprotect in sucrose. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 email pruegg@ihctech.net website www.ihctech.net IHC Resource Group www.ihcrg.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of TF Sent: Friday, May 15, 2009 12:15 PM To: Geoff McAuliffe Cc: Histonet@lists.utsouthwestern.edu Subject: Re: Re: [Histonet] 70% alcohol fixation of brain thanks to the reply. some antigens will be damaged by PFA fixation and can not be retreieval. the shock frozen section with acetone is of quite bad tissue quality. therefore I am trying alcohol! 2009-05-16 TF ???? Geoff McAuliffe ????? 2009-05-15 23:55:15 ???? tifei ??? Histonet@lists.utsouthwestern.edu ??? Re: [Histonet] 70% alcohol fixation of brain Greetings TF: Good luck freezing a brain that is full of alcohol! Have you checked the freezing point of alcohol? Why are you doing this? Immersing a whole brain in 70% alcohol, why?? 30% sucrose is a cryoprotectant so the brain is not full of holes from ice crystals.Alcohol defeats this purpose. Geoff TF wrote: > Hi, i dont want to use PFA for the brain fixation (rat). > Now I tried to perfuse the rat with saline, followed with 70% alcohol. Then I do post-fixation at room temperature for 24 hours. > I also tried saline perfusion, then I directly put the whole brain into 70% alcohol. > Is this fine? > > Also, for dehydration before cutting frozen sections on a cryostat/microtome, should I use 95% alcohol? I am now using 30% sucrose~ > > 2009-05-15 > > > > TF > _______________________________________________ > 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 mcauliff@umdnj.edu ********************************************** From gu.lang <@t> gmx.at Sat May 16 13:00:13 2009 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat May 16 13:00:17 2009 Subject: AW: [Histonet] Chemicals that inactivate the primary antibody In-Reply-To: <200905170048141643286@foxmail.com> References: <200905170048141643286@foxmail.com> Message-ID: For doublestaining the primary undergoes denaturation through a second HIER-step. The second secondary ab doesn't bind to the first primary. Therefore the binding sites must have been destroyed by heat in retrieval-buffer. Gudrun -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von TF Gesendet: Samstag, 16. Mai 2009 18:48 An: Histonet@lists.utsouthwestern.edu Betreff: [Histonet] Chemicals that inactivate the primary antibody Hi all, just wonder what kind of treatments/chemicals can complete block the binding of 2nd antibody to binded primary antibody on antigen? I tried HCl , but does not damage all the primaryantibody binding sites - i can still see staining pattern finally. 2009-05-17 TF _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tifei <@t> foxmail.com Sat May 16 13:16:57 2009 From: tifei <@t> foxmail.com (=?utf-8?B?VEY=?=) Date: Sat May 16 13:19:06 2009 Subject: =?utf-8?B?UmU6IEFXOiBbSGlzdG9uZXRdIENoZW1pY2FscyB0aGF0IGluYWN0aXZhdGUgdGhlIHByaW1hcnkgYW50aWJvZHk=?= References: <200905170048141643286@foxmail.com>, Message-ID: <200905170216518708740@foxmail.com> But the heat also damage the fluorescnece... u need specific amplication kit anyway. 2009-05-17 TF ???? Gudrun Lang ????? 2009-05-17 02:00:18 ???? tifei@foxmail.com ??? histonet@lists.utsouthwestern.edu ??? AW: [Histonet] Chemicals that inactivate the primary antibody For doublestaining the primary undergoes denaturation through a second HIER-step. The second secondary ab doesn't bind to the first primary. Therefore the binding sites must have been destroyed by heat in retrieval-buffer. Gudrun -----Urspr?gliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von TF Gesendet: Samstag, 16. Mai 2009 18:48 An: Histonet@lists.utsouthwestern.edu Betreff: [Histonet] Chemicals that inactivate the primary antibody Hi all, just wonder what kind of treatments/chemicals can complete block the binding of 2nd antibody to binded primary antibody on antigen? I tried HCl , but does not damage all the primaryantibody binding sites - i can still see staining pattern finally. 2009-05-17 TF _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From angrodoronar <@t> gmail.com Sun May 17 13:06:33 2009 From: angrodoronar <@t> gmail.com (Mark H) Date: Sun May 17 13:06:38 2009 Subject: [Histonet] Looking for employment in Tucson or Phoenix AZ Message-ID: <16c76c910905171106h33a08a1asbc8e4f090df7cd9c@mail.gmail.com> I?m a recent Arizona Pima Community College graduate in histology seeking full time employment in either the Tucson or Phoenix area. Current member of ASH and NSH, as well as 1 year experience in co-ops at Ventana Medical Systems and North West Medical Center in Tucson. Mark 520-904-8005 or respond to this E-mail Thank You From mari_ferrarini <@t> hotmail.com Sun May 17 15:04:38 2009 From: mari_ferrarini <@t> hotmail.com (Mari Ferrarini) Date: Sun May 17 15:04:42 2009 Subject: [Histonet] TRAP and ALP staining Message-ID: Hello all, I'm currently trying to do TRAP and ALP staining in MMA (Technovit 9100) and GMA (cold embedding) embedded sections. The thing is I don't have a lot of experience and don't really know what could go wrong. I'm doing the stainings proposed by Chappard and Liu for TRAP and the staining proposed by Liu for ALP, as follows: TRAP LIU: (1 mL : 1 mL) 4% NaNO2 + 4% Pararosaniline; 23 mL 0,1M Acetate Buffer; 8 mg Naphtol AS-TR Phosphate 0,3 mL Dimethylformamide; 57,5 mg Sodium Tartrate pH: 5; 37oC/1 h; ALP LIU: 24 mg Fast Blue BB 30 mL 0,1M Tris Buffer; 8 mg naphtol AS-TR Phosphate 0,3 mL Dimethylformamide 10 mg MgCl2 pH: 9; 37oC/30 min; TRAP CHAPPARD: 100 mg Sodium alfa-naphtyl phosphate 200 mg Fast Violet B; 15 mg Sodium Tartrate; 100 mL 0,1M Acetate Buffer, pH 5; 37oC/30 min Is there a problem if I change the phosphate? I only have a naphthol AS-MX phosphate. Is it equivalent for these procedures? I'm not really sure either on the kind of mounting media I'm supposed to use. Aqueous mounting for both MMA and GMA? I'm also not sure of how to attach the sections to the slides prior to staining in a way that doesn't affect much the enzyme activity. Thank you very much for the time and patience, any help would be appreciated. Sincerely, Mariana Ferrarini Lyon, France. _________________________________________________________________ Emoticons e Winks super diferentes para o Messenger. Baixe agora, ? gr?tis! http://specials.br.msn.com/ilovemessenger/pacotes.aspx From zafarparkview <@t> yahoo.com Mon May 18 06:16:07 2009 From: zafarparkview <@t> yahoo.com (shama zafar) Date: Mon May 18 06:16:17 2009 Subject: [Histonet] Immunohistochemistry Message-ID: <704480.95667.qm@web55601.mail.re4.yahoo.com> I am working in immuno Lab doing hgh volume and i need resource where i can have some continuing education.Thanks if any one can help. From mtighe <@t> trudeauinstitute.org Mon May 18 08:08:25 2009 From: mtighe <@t> trudeauinstitute.org (Mike Tighe) Date: Mon May 18 08:08:37 2009 Subject: [Histonet] Cytoseal XYL Message-ID: <4A112585.26E4.00EE.0@trudeauinstitute.org> Has anyone compared Cytoseal 60 with Cytoseal XYL? I clear with Xylene and use cytoseal 60 (Toluene)to coverslip. Seems like it would make sense to use Cytoseal XYL (Xylene) but they only sell in a case and I can't get a sample. I would be interested to know if anyone has a preference. Thanks!! Mike From leiker <@t> buffalo.edu Mon May 18 08:42:30 2009 From: leiker <@t> buffalo.edu (Merced M Leiker) Date: Mon May 18 08:42:36 2009 Subject: AW: [Histonet] Chemicals that inactivate the primary antibody In-Reply-To: <200905170216518708740@foxmail.com> References: <200905170048141643286@foxmail.com> , <200905170216518708740@foxmail.com> Message-ID: <553BB3DB58FAE828B383F988@CDYwxp1931.ad.med.buffalo.edu> Hi TF and all interested, I think I know what you want, but unfortunately I don't know how to answer your question (it is something I'd like answered myself!!) To re-word for the sake of all interested: You want to perform double-immunofluorescent staining using 2 primaries that were raised in the same species. An additional question for clarification is: Do you want to do this on paraffin or frozen sections? Maybe someone can help figure it out... Merced --On Sunday, May 17, 2009 2:16 AM +0800 TF wrote: > But the heat also damage the fluorescnece... > u need specific amplication kit anyway. > > > 2009-05-17 > > > > TF > > > > ???? Gudrun Lang > ????? 2009-05-17 02:00:18 > ???? tifei@foxmail.com > ??? histonet@lists.utsouthwestern.edu > ??? AW: [Histonet] Chemicals that inactivate the primary antibody > > For doublestaining the primary undergoes denaturation through a second > HIER-step. The second secondary ab doesn't bind to the first primary. > Therefore the binding sites must have been destroyed by heat in > retrieval-buffer. > Gudrun > -----Urspr?gliche Nachricht----- > Von: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von TF > Gesendet: Samstag, 16. Mai 2009 18:48 > An: Histonet@lists.utsouthwestern.edu > Betreff: [Histonet] Chemicals that inactivate the primary antibody > Hi all, just wonder what kind of treatments/chemicals can complete block > the binding of 2nd antibody to binded primary antibody on antigen? > I tried HCl , but does not damage all the primaryantibody binding sites - > i can still see staining pattern finally. > 2009-05-17 > TF > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 leiker@buffalo.edu 716-829-6118 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. From sjohnson <@t> neogenomics.org Fri May 8 12:53:36 2009 From: sjohnson <@t> neogenomics.org (Saundra Johnson) Date: Mon May 18 09:25:47 2009 Subject: [Histonet] Slide quality Message-ID: Hi Bernice, I noticed a post in the histonet archives about using ammonia water to soak. Could I ask your opinion about how ammonia water could effect the IHC staining? I have come to work here and the two techs here soak in 50% ammonia water. That seems way to strong to me. What do you think? Saundra Johnson, HT(ASCP)QIHC Histology Supervisor NeoGenomics Laboratories "When Time Matters...And Results Count" 12701 Commonwealth Drive, Suite 9 Fort Myers, FL 33913 Phone: (239) 768-0600 Ext. 2229 Fax: (239) 768-0711 ________________________________ This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. E-mail transmission cannot be guaranteed to be secured or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The sender therefore does not accept liability for any errors or omissions in the contents of this message, which arise as a result of e-mail transmission. If verification is required please request a hard-copy version. NeoGenomics Laboratories, Suite 5, 12701 Commonwealth Dr, Fort Myers, FL 33913, http://www.neogenomics.org From nmac <@t> staffmail.ed.ac.uk Mon May 11 07:23:12 2009 From: nmac <@t> staffmail.ed.ac.uk (Neil Macintyre) Date: Mon May 18 09:25:49 2009 Subject: [Histonet] Canine CD25 Message-ID: <20090511132312362.00000002372@VCS-127075> Hi everyone. I have a researcher that would like to look at CD25 in canine tissue. Has anyone ever used this antibody? Any information would be much appreciated Neil MacIntyre CSci FIBMS Laboratory Manager Veterinary Pathology Unit Easter Bush Veterinary Centre Nr Roslin Midlothian EH25 9RG phone:0131 650 6403/8802 Fax: 0131 445 5770 The University of Edinburgh is a charitable body, registered in Scotland, with registration number SC005336. From LMurphy2 <@t> aultman.com Mon May 11 09:33:01 2009 From: LMurphy2 <@t> aultman.com (Leann M. Murphy) Date: Mon May 18 09:25:52 2009 Subject: [Histonet] Thermo-Fisher Products Message-ID: <6DD7EFAF15E66C4583A82119CAF1590C0A449A3E@exch04.ahf2000.aultman.com> I was just wondering what everyone's thoughts were on thermo products. I have been demo-ing an Exlcelsior Processor and was wondering if anyone had any problems. Let me know. LeAnn Murphy From nmac <@t> staffmail.ed.ac.uk Wed May 13 11:15:03 2009 From: nmac <@t> staffmail.ed.ac.uk (Neil Macintyre) Date: Mon May 18 09:25:53 2009 Subject: [Histonet] CD25 Message-ID: <20090513171503042.00000003196@VCS-127075> Hi everyone. I have a researcher that would like to look at CD25 in canine tissue. Has anyone ever used this antibody? Any information would be much appreciated Neil MacIntyre CSci FIBMS Laboratory Manager Veterinary Pathology Unit Easter Bush Veterinary Centre Nr Roslin Midlothian EH25 9RG phone:0131 650 6403/8802 Fax: 0131 445 5770 The University of Edinburgh is a charitable body, registered in Scotland, with registration number SC005336. From Karen.Kay <@t> albertahealthservices.ca Wed May 13 12:55:49 2009 From: Karen.Kay <@t> albertahealthservices.ca (Karen Kay) Date: Mon May 18 09:25:55 2009 Subject: [Histonet] DAB WASTE In-Reply-To: References: Message-ID: Hello Jennifer, We are a hospital site and operate 2 Dako Autostainers. We collect our DAB waste into empty plastic carboys from Coulter Diluent used in our Hematology Department. These containers are encased within a cardboard cover. They are very convenient as we can place the hose directly into the waste container from the Dako itself. Our volume is such that we discard the containers on a weekly basis. Hope this information is useful to you Karen K Histopathology Dept Lethbridge Regional Hospital Lethbridge,Alberta, CANADA ------------------------------ Message: 6 Date: Wed, 13 May 2009 05:51:33 -0700 From: "Jennifer Campbell" Subject: [Histonet] Question about DAB waste To: Message-ID: <5658CBDB9EAE6545ABE50D2563D81BF821CFC4@VDXSERVER01.vdxpathology.local> Content-Type: text/plain; charset="US-ASCII" We just purchased a Dako Autostainer and are in the process of figuring out how to get rid of the haz waste that we will be generating. The machine separates the haz from non-haz, thankfully. I've been communicating with our waste facility and have been having some difficulty. They want to know a lot of specifics like, how much waste we will be brining to them at a time and how often, what sort of containers we will be bringing it in etc. Do you let the haz waste carboy get to a certain level before emptying it? Is it ok to pour this waste into plastic containers of some sort (we were going to use empty distilled water bottles) to transport them to the facility? I would appreciate any insight. Thank you. Jennifer C. This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. From ddiaz <@t> mercedesmedical.com Fri May 15 12:38:04 2009 From: ddiaz <@t> mercedesmedical.com (Dan Diaz) Date: Mon May 18 09:25:57 2009 Subject: [Histonet] KP Marker Plus In-Reply-To: References: Message-ID: That is Mercedes Medicals LIST Price for those accounts that order online and order 1 box per year. If you order through a Histology Sales Rep (800-331-2716) You may enjoy a discounted price. Because Mercedes Medical Reps are nice and appreciate great customers! -----Original Message----- From: Dave Johnson Sent: Friday, May 15, 2009 1:34 PM To: Sales Inside; Marketing Department Subject: FW: [Histonet] KP Marker Plus Theres some great free marketing. Anyone want to couthly respond? Or maybe lower our list price. We don't sell to that many at that price Whose? SUNY Buffalo , asses -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Merced M Leiker Sent: Friday, May 15, 2009 1:23 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] KP Marker Plus By the way, they are only $55/box... :-) --On Friday, May 15, 2009 1:17 PM -0400 Merced M Leiker wrote: > Indeed! My box of 12 pens just arrived today. Cat# KLI KPPEN. > www.mercedesmedical.com > > Merced > > --On Friday, May 15, 2009 11:16 AM -0500 "Molinari, Betsy" > wrote: > >> Mercedes Medical 800-331-2716 >> >> Betsy Molinari HT (ASCP) >> Texas heart Institute >> Cardiovascular Pathology >> Houston,TX 77090 >> 832-355-6524 >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >> Metzger, Kenneth >> Sent: Friday, May 15, 2009 11:13 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] KP Marker Plus >> >> Does anyone know who sells the KP Marker PLUS? I can't seem to find >> them. Thanks for any help. >> >> >> >> Ken >> >> >> >> Kenneth G Metzger HTL(ASCP) >> >> ARUP Labs >> >> Histology Supervisor >> >> 500 Chipeta Way >> >> Salt Lake City, Utah 84108-1221 >> >> kenneth.metzger@aruplab.com >> >> (801) 583-2787 x 3101 >> >> >> >> >> - ------------------------------------------------------------------ >> The information transmitted by this e-mail and any included >> attachments are from ARUP Laboratories and are intended only for the >> recipient. The information contained in this message is confidential >> and may constitute inside or non-public information under >> international, federal, or state securities laws, or protected health >> information and is intended only for the use of the recipient. >> Unauthorized forwarding, printing, copying, distributing, or use of >> such information is strictly prohibited and may be unlawful. If you >> are not the intended recipient, please promptly delete this e-mail >> and notify the sender of the delivery error or you may call ARUP >> Laboratories Compliance Hot Line in Salt Lake City, Utah USA at (+1 >> (800) 522-2787 ext. 2100 >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > Merced M Leiker > Cardiovascular Medicine > 348 Biomedical Research Building > State University of New York at Buffalo > 3435 Main St, Buffalo, NY 14214 > leiker@buffalo.edu > 716-829-6118 > > No trees were harmed in the sending of this email. > However, many electrons were severely inconvenienced. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 leiker@buffalo.edu 716-829-6118 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon May 18 09:31:22 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon May 18 09:31:25 2009 Subject: AW: [Histonet] Chemicals that inactivate the primary antibody Message-ID: <261542.82019.qm@web65705.mail.ac4.yahoo.com> I am completely confused. IF you have an epitope in the tissue and you react with it an specific antibody "that is it". The epitope will have reacted and I do not see any way in which you can add another antibody to that initial epitope. From the reactive point of view, the epitope is "blocked" to react with another, unless the reaction is not totally specific and "there is room for another". I just cannot imagine a way of doing this. Ren? J. --- On Mon, 5/18/09, Merced M Leiker wrote: From: Merced M Leiker Subject: Re: AW: [Histonet] Chemicals that inactivate the primary antibody To: tifei@foxmail.com, gu.lang@gmx.at Cc: histonet@lists.utsouthwestern.edu Date: Monday, May 18, 2009, 9:42 AM Hi TF and all interested, I think I know what you want, but unfortunately I don't know how to answer your question (it is something I'd like answered myself!!)? To re-word for the sake of all interested: You want to perform double-immunofluorescent staining using 2 primaries that were raised in the same species. An additional question for clarification is: Do you want to do this on paraffin or frozen sections? Maybe someone can help figure it out... Merced --On Sunday, May 17, 2009 2:16 AM +0800 TF wrote: > But the heat also damage the fluorescnece... > u need specific amplication kit anyway. > > > 2009-05-17 > > > > TF > > > > ???? Gudrun Lang > ????? 2009-05-17? 02:00:18 > ???? tifei@foxmail.com > ??? histonet@lists.utsouthwestern.edu > ??? AW: [Histonet] Chemicals that inactivate the primary antibody > > For doublestaining the primary undergoes denaturation through a second > HIER-step. The second secondary ab doesn't bind to the first primary. > Therefore the binding sites must have been destroyed by heat in > retrieval-buffer. > Gudrun > -----Urspr?gliche Nachricht----- > Von: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von TF > Gesendet: Samstag, 16. Mai 2009 18:48 > An: Histonet@lists.utsouthwestern.edu > Betreff: [Histonet] Chemicals that inactivate the primary antibody > Hi all, just wonder what kind of treatments/chemicals can complete block > the binding of 2nd antibody to binded primary antibody on antigen? > I tried HCl , but does not damage all the primaryantibody binding sites - > i can still see staining pattern finally. > 2009-05-17 > TF > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY? 14214 leiker@buffalo.edu 716-829-6118 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cbrya <@t> lexclin.com Mon May 18 09:41:07 2009 From: cbrya <@t> lexclin.com (Carol Bryant) Date: Mon May 18 09:41:22 2009 Subject: [Histonet] Thermo-Fisher Products In-Reply-To: <6DD7EFAF15E66C4583A82119CAF1590C0A449A3E@exch04.ahf2000.aultman.com> References: <6DD7EFAF15E66C4583A82119CAF1590C0A449A3E@exch04.ahf2000.aultman.com> Message-ID: <37A1F9CAB9E21C41B39F3653B620D13E093BA296@exchange2003.lc.local> We have an Excelsior that is about 5 years old. We have had a lot of issues and have had service too many times in the past year to count. We are looking at replacing it. Perhaps we got a lemon, but I don't think I could recommend it based on our experience. Are there any other processors anyone would like to recommend? Also, how about microwave processors and IHC? Thanks, Carol -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leann M. Murphy Sent: Monday, May 11, 2009 10:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Thermo-Fisher Products I was just wondering what everyone's thoughts were on thermo products. I have been demo-ing an Exlcelsior Processor and was wondering if anyone had any problems. Let me know. LeAnn Murphy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by the State of Kentucky and/or Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient and have received this communication in error, please contact the sender and destroy all copies of the original message. Thank you. From tifei <@t> foxmail.com Mon May 18 10:37:19 2009 From: tifei <@t> foxmail.com (TF) Date: Mon May 18 10:37:42 2009 Subject: =?utf-8?B?UmU6IFJlOiBBVzogW0hpc3RvbmV0XSBDaGVtaWNhbHMgdGhhdCBpbmFjdGl2YXRlIHRoZSBwcmltYXJ5IGFudGlib2R5?= References: <261542.82019.qm@web65705.mail.ac4.yahoo.com> Message-ID: <200905182337139539275@foxmail.com> Thanks all, Merced M Leiker is right: the double IHC procedure using two primary antibodies from same species. Some expert just sent me their procedure of Heat-interval based inactivation of the primary antibody for first antigen. I am just seeking for a chemical way at room temperature. 2009-05-18 TF ???? Rene J Buesa ????? 2009-05-18 22:31:30 ???? tifei; gu.lang; Merced M Leiker ??? histonet ??? Re: AW: [Histonet] Chemicals that inactivate the primary antibody I am completely confused. IF you have an epitope in the tissue and you react with it an specific antibody "that is it". The epitope will have reacted and I do not see any way in which you can add another antibody to that initial epitope. From the reactive point of view, the epitope is "blocked" to react with another, unless the reaction is not totally specific and "there is room for another". I just cannot imagine a way of doing this. Ren? J. --- On Mon, 5/18/09, Merced M Leiker wrote: From: Merced M Leiker Subject: Re: AW: [Histonet] Chemicals that inactivate the primary antibody To: tifei@foxmail.com, gu.lang@gmx.at Cc: histonet@lists.utsouthwestern.edu Date: Monday, May 18, 2009, 9:42 AM Hi TF and all interested, I think I know what you want, but unfortunately I don't know how to answer your question (it is something I'd like answered myself!!) To re-word for the sake of all interested: You want to perform double-immunofluorescent staining using 2 primaries that were raised in the same species. An additional question for clarification is: Do you want to do this on paraffin or frozen sections? Maybe someone can help figure it out... Merced --On Sunday, May 17, 2009 2:16 AM +0800 TF wrote: > But the heat also damage the fluorescnece... > u need specific amplication kit anyway. > > > 2009-05-17 > > > > TF > > > > ???? Gudrun Lang > ????? 2009-05-17 02:00:18 > ???? tifei@foxmail.com > ??? histonet@lists.utsouthwestern.edu > ??? AW: [Histonet] Chemicals that inactivate the primary antibody > > For doublestaining the primary undergoes denaturation through a second > HIER-step. The second secondary ab doesn't bind to the first primary. > Therefore the binding sites must have been destroyed by heat in > retrieval-buffer. > Gudrun > -----Urspr?gliche Nachricht----- > Von: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von TF > Gesendet: Samstag, 16. Mai 2009 18:48 > An: Histonet@lists.utsouthwestern.edu > Betreff: [Histonet] Chemicals that inactivate the primary antibody > Hi all, just wonder what kind of treatments/chemicals can complete block > the binding of 2nd antibody to binded primary antibody on antigen? > I tried HCl , but does not damage all the primaryantibody binding sites - > i can still see staining pattern finally. > 2009-05-17 > TF > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 leiker@buffalo.edu 716-829-6118 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LGarrison <@t> neogenomics.org Mon May 18 10:51:35 2009 From: LGarrison <@t> neogenomics.org (Lindsey Garrison) Date: Mon May 18 10:51:49 2009 Subject: [Histonet] control tissue Message-ID: <565D7CED3531984A84D9D80EB90C48783E06680C53@aneuploidy.neogen.local> We have a plentiful amount (hundreds) of malignat melanoma blocks that we are willing to trade some for any ER/PR + tissue, colon cancer, or any other uselful IHC control tissue. Any takers? ________________________________ This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. E-mail transmission cannot be guaranteed to be secured or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The sender therefore does not accept liability for any errors or omissions in the contents of this message, which arise as a result of e-mail transmission. If verification is required please request a hard-copy version. NeoGenomics Laboratories, Suite 5, 12701 Commonwealth Dr, Fort Myers, FL 33913, http://www.neogenomics.org From Timothy.Morken <@t> ucsfmedctr.org Mon May 18 11:08:14 2009 From: Timothy.Morken <@t> ucsfmedctr.org (Morken, Tim) Date: Mon May 18 11:08:26 2009 Subject: FW: Re: [Histonet] Chemicals that inactivate the primary antibody Message-ID: <1AAF670737F193429070841C6B2ADD4C01E54EAE@EXMBMCB15.ucsfmedicalcenter.org> DQoNClRGLCANCg0KVHdvIHRoaW5ncyB5b3UgY2FuIGRvOw0KDQpJZiB5b3UgYXJlIHVzaW5nIGEg Y2hyb21vZ2VuaWMgc3lzdGVtIChub3QgZmx1b3Jlc2NlbmNlKSB5b3UgY2FuIHVzZSBEQUIgYXMg dGhlIGZpcnN0IGNocm9tb2dlbi4gVGhlIERBQiBwb2x5bWVyaXplZCBwcm9kdWN0IHdpbGwgY292 ZXIgdGhlIGZpcnN0IHByaW1hcnkgYW5kIHByZXZlbnQgY3Jvc3MtcmVhY3Rpb24uDQoNCk9yLCBp ZiB1c2luZyBJRiB5b3UgY2FuIHVzZSB1bmxhYmxlZCBGYWIgZnJhZ21lbnRzIHRvIGNvYXQgdGhl IGZpcnN0IHByaW1hcnkuIEZvciBpbnN0YW5jZSwgdXNlIGRvbmtleSBhbnRpLW1vdXNlIHRvIGNv YXQgdGhlIGZpcnN0IHByaW1hcnkgKGlmIGl0IGlzIGEgbW91c2UgYW50aWJvZHkpLiBTZWUgSmFj a3NvbiBJbW11bm8gUmVzZWFyY2ggZm9yIHRoZSBhbnRpYm9keSBmcmFnbWVudHMuDQoNClNlZSB0 aGlzIGxpbmsgZm9yIGZ1bGwgaW5mb3JtYXRpb246ICBodHRwOi8vd3d3LmphY2tzb25pbW11bm8u Y29tL3RlY2huaWNhbC9mYWItYmxvay5hc3ANCg0KVGltIE1vcmtlbg0KU3VwZXJ2aXNvciwgSGlz dG9sb2d5IC8gSVBPWA0KVUNTRiBNZWRpY2FsIENlbnRlcg0KU2FuIEZyYW5jaXNjbywgQ0EgIA0K IA0KDQotLS0tLU9yaWdpbmFsIE1lc3NhZ2UtLS0tLQ0KRnJvbTogaGlzdG9uZXQtYm91bmNlc0Bs aXN0cy51dHNvdXRod2VzdGVybi5lZHUgW21haWx0bzpoaXN0b25ldC1ib3VuY2VzQGxpc3RzLnV0 c291dGh3ZXN0ZXJuLmVkdV0gT24gQmVoYWxmIE9mIFRGDQpTZW50OiBTYXR1cmRheSwgTWF5IDE2 LCAyMDA5IDEwOjM4IEFNDQpUbzogUmVuZSBKIEJ1ZXNhOyBIaXN0b25ldEBsaXN0cy51dHNvdXRo d2VzdGVybi5lZHUNClN1YmplY3Q6IFJlOiBSZTogW0hpc3RvbmV0XSBDaGVtaWNhbHMgdGhhdCBp bmFjdGl2YXRlIHRoZSBwcmltYXJ5IGFudGlib2R5DQoNCkhpLCBidXQgSSBkb250IHdhbnQgdG8g ZGVzdG9yeSBldmVyeXRoaW5nLi4uLg0KSSBhbSB0cnlpbmcgdG8gc2VlIGlmIEkgY2FuIHByZXZl bnQgdGhlIGJpbmRpbmcgb2YgYW5vdGhlciAybmQgYW50aWJvZHkgdG8gdGhlIHByaW1hcnkgYW50 aWJvZHkgZm9yIHRoZSBmaXJzdCBhbnRpZ2VuIQ0KVGhlcmUgZm9yZSB5b3UgY2FuIHBlcmZvcm0g ZG91YmxlIHN0YWluaW5nIGZyb20gc2FtZSBzZXBlY2llcy1kZXJpdmVkIHByaW1hcnkgYW50aWJv ZGllcyENCg0KDQoyMDA5LTA1LTE3IA0KDQoNCg0KVEYgDQoNCg0KDQq3orz+yMujuiBSZW5lIEog QnVlc2EgDQq3osvNyrG85KO6IDIwMDktMDUtMTcgIDAwOjU0OjI2IA0KytW8/sjLo7ogSGlzdG9u ZXRAbGlzdHMudXRzb3V0aHdlc3Rlcm4uZWR1OyB0aWZlaSANCrOty82juiANCtb3zOKjuiBSZTog W0hpc3RvbmV0XSBDaGVtaWNhbHMgdGhhdCBpbmFjdGl2YXRlIHRoZSBwcmltYXJ5IGFudGlib2R5 IA0KIA0KVHJ5IGJsZWFjaCBhbmQgeW91IHdpbGwgZGVzdHJveSBldmVyeXRoaW5nIQ0KUmVuqKYg Si4NCg0KLS0tIE9uIFNhdCwgNS8xNi8wOSwgVEYgPHRpZmVpQGZveG1haWwuY29tPiB3cm90ZToN Cg0KDQpGcm9tOiBURiA8dGlmZWlAZm94bWFpbC5jb20+DQpTdWJqZWN0OiBbSGlzdG9uZXRdIENo ZW1pY2FscyB0aGF0IGluYWN0aXZhdGUgdGhlIHByaW1hcnkgYW50aWJvZHkNClRvOiAiSGlzdG9u ZXRAbGlzdHMudXRzb3V0aHdlc3Rlcm4uZWR1IiA8SGlzdG9uZXRAbGlzdHMudXRzb3V0aHdlc3Rl cm4uZWR1Pg0KRGF0ZTogU2F0dXJkYXksIE1heSAxNiwgMjAwOSwgMTI6NDggUE0NCg0KDQpIaSBh bGwsIGp1c3Qgd29uZGVyIHdoYXQga2luZCBvZiB0cmVhdG1lbnRzL2NoZW1pY2FscyBjYW4gY29t cGxldGUgYmxvY2sgdGhlIGJpbmRpbmcgb2YgMm5kIGFudGlib2R5IHRvIGJpbmRlZCBwcmltYXJ5 IGFudGlib2R5IG9uIGFudGlnZW4/DQpJIHRyaWVkIEhDbCAsIGJ1dCBkb2VzIG5vdCBkYW1hZ2Ug YWxsIHRoZSBwcmltYXJ5YW50aWJvZHkgYmluZGluZyBzaXRlcyAtIGkgY2FuIHN0aWxsIHNlZSBz dGFpbmluZyBwYXR0ZXJuIGZpbmFsbHkuDQoNCjIwMDktMDUtMTcgDQoNCg0KDQpURiANCl9fX19f X19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fDQpIaXN0b25ldCBtYWls aW5nIGxpc3QNCkhpc3RvbmV0QGxpc3RzLnV0c291dGh3ZXN0ZXJuLmVkdQ0KaHR0cDovL2xpc3Rz LnV0c291dGh3ZXN0ZXJuLmVkdS9tYWlsbWFuL2xpc3RpbmZvL2hpc3RvbmV0DQo= From algranth <@t> email.arizona.edu Mon May 18 11:13:45 2009 From: algranth <@t> email.arizona.edu (Andrea Grantham) Date: Mon May 18 11:13:52 2009 Subject: [Histonet] coverslipping helper Message-ID: Good Monday morning! This may sound crazy...or not. I have to admit that I have never heard of this device before and I was wondering if any of you out in histoland could tell me what this is and where one might go to get it, if it were available for sale. Here goes: A student who was having a difficult time learning how to coverslip slides in my lab told me that she used to work for a doctor who invented a device to help coverslip. It was "like a magnet" and just eliminated all bubbles from under the coverslip. Anybody know? Andi Andrea Grantham, HT (ASCP) Senior Research Specialist University of Arizona Cell Biology and Anatomy Histology Service Laboratory P.O.Box 245044 Tucson, AZ 85724 algranth@email.arizona.edu Tel: 520.626.4415 Fax: 520.626.2097 "happy slicing and dicing and may all your stains work perfectly" - Paula Sicurello From nefff <@t> staff.uni-marburg.de Mon May 18 11:16:51 2009 From: nefff <@t> staff.uni-marburg.de (Dr. Frauke Neff) Date: Mon May 18 11:17:20 2009 Subject: AW: [Histonet] Chemicals that inactivate the primary antibody In-Reply-To: <200905182337139539275@foxmail.com> References: <261542.82019.qm@web65705.mail.ac4.yahoo.com> <200905182337139539275@foxmail.com> Message-ID: <20090518181651.tx8fvdph6owks4cw@home.staff.uni-marburg.de> Hi TF, we do IF with two 1.ABs out of mice using a sequential protocol from abcam: www.abcam.com/technical We perform a second blocking step with serum (higher conc. than the first one) after the first Fluorescence was added, do not perform another retrieval procedure (my ABs need the same) and perform the following incubation steps in the "dark" (we switch the light off and cover the slides after adding the ABs) to prevent fading. Until now, I was sure, this procedure works but maybe the two antigens don't colocalize but it is just a staining artifact ;-) Hope this helps, Frauke Zitat von TF : > Thanks all, Merced M Leiker is right: the double IHC procedure using > two primary antibodies from same species. > > Some expert just sent me their procedure of Heat-interval based > inactivation of the primary antibody for first antigen. > I am just seeking for a chemical way at room temperature. > > > 2009-05-18 > > > > TF > > > > ???? Rene J Buesa > ????? 2009-05-18 22:31:30 > ???? tifei; gu.lang; Merced M Leiker > ??? histonet > ??? Re: AW: [Histonet] Chemicals that inactivate the primary antibody > > I am completely confused. > IF you have an epitope in the tissue and you react with it an > specific antibody "that is it". The epitope will have reacted and I > do not see any way in which you can add another antibody to that > initial epitope. From the reactive point of view, the epitope is > "blocked" to react with another, unless the reaction is not totally > specific and "there is room for another". > I just cannot imagine a way of doing this. > Ren? J. > > --- On Mon, 5/18/09, Merced M Leiker wrote: > > > From: Merced M Leiker > Subject: Re: AW: [Histonet] Chemicals that inactivate the primary antibody > To: tifei@foxmail.com, gu.lang@gmx.at > Cc: histonet@lists.utsouthwestern.edu > Date: Monday, May 18, 2009, 9:42 AM > > > Hi TF and all interested, > > I think I know what you want, but unfortunately I don't know how to answer > your question (it is something I'd like answered myself!!) To re-word for > the sake of all interested: > > You want to perform double-immunofluorescent staining using 2 primaries > that were raised in the same species. > > An additional question for clarification is: Do you want to do this on > paraffin or frozen sections? > > Maybe someone can help figure it out... > > Merced > > > --On Sunday, May 17, 2009 2:16 AM +0800 TF wrote: > >> But the heat also damage the fluorescnece... >> u need specific amplication kit anyway. >> >> >> 2009-05-17 >> >> >> >> TF >> >> >> >> ???? Gudrun Lang >> ????? 2009-05-17 02:00:18 >> ???? tifei@foxmail.com >> ??? histonet@lists.utsouthwestern.edu >> ??? AW: [Histonet] Chemicals that inactivate the primary antibody >> >> For doublestaining the primary undergoes denaturation through a second >> HIER-step. The second secondary ab doesn't bind to the first primary. >> Therefore the binding sites must have been destroyed by heat in >> retrieval-buffer. >> Gudrun >> -----Urspr?gliche Nachricht----- >> Von: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von TF >> Gesendet: Samstag, 16. Mai 2009 18:48 >> An: Histonet@lists.utsouthwestern.edu >> Betreff: [Histonet] Chemicals that inactivate the primary antibody >> Hi all, just wonder what kind of treatments/chemicals can complete block >> the binding of 2nd antibody to binded primary antibody on antigen? >> I tried HCl , but does not damage all the primaryantibody binding sites - >> i can still see staining pattern finally. >> 2009-05-17 >> TF >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > Merced M Leiker > Cardiovascular Medicine > 348 Biomedical Research Building > State University of New York at Buffalo > 3435 Main St, Buffalo, NY 14214 > leiker@buffalo.edu > 716-829-6118 > > No trees were harmed in the sending of this email. > However, many electrons were severely inconvenienced. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Jeannette.Wallen <@t> hcmed.org Mon May 18 11:29:25 2009 From: Jeannette.Wallen <@t> hcmed.org (Wallen, Jeannette B) Date: Mon May 18 11:29:32 2009 Subject: [Histonet] Steel Microtome Blades Message-ID: <66FFB482DE2B624DA30B826C1795382148E4E844FD@MCD-MBX-3.HCMC.CO.HENNEPIN.MN.US> Does anyone still use the steel microtome blades that need to be sharpened? We have found some of these blades in our laboratory and will dispose of them, if no one has any use for them. Jeannette B. Wallen, HT (ASCP) Histology Specialist/Histology Team Lead Hennepin County Medical Center Anatomic Pathology/Histology Lab 701 Park Avenue, Mail Code: PL Minneapolis, MN 55415 (612) 873-9108 Office, PL.731 (612) 510-5677 Pager (612) 873-3079 General Histology (612) 904-4629 Fax Email Address: Jeannette.Wallen@hcmed.org From histonet.nospam <@t> vneubert.com Mon May 18 11:41:28 2009 From: histonet.nospam <@t> vneubert.com (V. Neubert) Date: Mon May 18 11:41:36 2009 Subject: [Histonet] Cytoseal XYL In-Reply-To: <4A112585.26E4.00EE.0@trudeauinstitute.org> References: <4A112585.26E4.00EE.0@trudeauinstitute.org> Message-ID: <4A118FB8.8040605@vneubert.com> I have not compared 60 to XYL but started with XYL from the beginning. I use n-butyle acetate (http://en.wikipedia.org/wiki/Butyl_acetate) for clearing, its MSDS is quite satisfying and it smells way better. Mike Tighe wrote: > Has anyone compared Cytoseal 60 with Cytoseal XYL? I clear with Xylene and use cytoseal 60 (Toluene)to coverslip. Seems like it would make sense to use Cytoseal XYL (Xylene) but they only sell in a case and I can't get a sample. I would be interested to know if anyone has a preference. > > Thanks!! > Mike > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From rjbuesa <@t> yahoo.com Mon May 18 12:05:36 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon May 18 12:05:38 2009 Subject: [Histonet] coverslipping helper Message-ID: <232678.60898.qm@web65706.mail.ac4.yahoo.com> Never heard of that, but perhaps she is referring to placing some small weight over the coverslipped section to help eliminate the bubbles by the weight, other than that, that is totally new for me. Ren? J. --- On Mon, 5/18/09, Andrea Grantham wrote: From: Andrea Grantham Subject: [Histonet] coverslipping helper To: "HISTONET" Date: Monday, May 18, 2009, 12:13 PM Good Monday morning! This may sound crazy...or not. I have to admit that I have never heard of this device before and I was wondering if any of you out in histoland could tell me what this is and where one might go to get it, if it were available for sale. Here goes: A student who was having a difficult time learning how to coverslip slides in my lab told me that she used to work for a doctor who invented a device to help coverslip. It was "like a magnet" and just eliminated all bubbles from under the coverslip. Anybody know? Andi Andrea Grantham, HT (ASCP) Senior Research Specialist University of Arizona Cell Biology and Anatomy Histology Service Laboratory P.O.Box 245044 Tucson, AZ 85724 algranth@email.arizona.edu Tel: 520.626.4415? ???Fax: 520.626.2097 "happy slicing and dicing and may all your stains work perfectly" - Paula Sicurello _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From estellamireles <@t> gmail.com Mon May 18 12:16:13 2009 From: estellamireles <@t> gmail.com (Stella Mireles) Date: Mon May 18 12:16:19 2009 Subject: [Histonet] Alpha-naphthyl-butyrate Message-ID: Looking for the order number for Alpha-naphthyl-butyrate. I will be using it for the NSE procedure. I spoke with the Fisher 800 number and they were unable to help me. We use Fisher or Sigma products. Thanks From esther.peters <@t> verizon.net Mon May 18 12:26:14 2009 From: esther.peters <@t> verizon.net (Esther Peters) Date: Mon May 18 12:26:32 2009 Subject: [Histonet] Steel Microtome Blades In-Reply-To: <66FFB482DE2B624DA30B826C1795382148E4E844FD@MCD-MBX-3.HCMC.CO.HENNEPIN.MN.US> References: <66FFB482DE2B624DA30B826C1795382148E4E844FD@MCD-MBX-3.HCMC.CO.HENNEPIN.MN.US> Message-ID: <4A119A36.5040001@verizon.net> I also have a number of these blades and an old knife sharpener, which I would like to get to a good home. Esther Peters George Mason University Wallen, Jeannette B wrote: > Does anyone still use the steel microtome blades that need to be sharpened? We have found some of these blades in our laboratory and will dispose of them, if no one has any use for them. > > Jeannette B. Wallen, HT (ASCP) > Histology Specialist/Histology Team Lead > Hennepin County Medical Center > Anatomic Pathology/Histology Lab > 701 Park Avenue, Mail Code: PL > Minneapolis, MN 55415 > (612) 873-9108 Office, PL.731 > (612) 510-5677 Pager > (612) 873-3079 General Histology > (612) 904-4629 Fax > Email Address: Jeannette.Wallen@hcmed.org > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From wdesalvo.cac <@t> hotmail.com Mon May 18 12:34:11 2009 From: wdesalvo.cac <@t> hotmail.com (WILLIAM DESALVO) Date: Mon May 18 12:34:15 2009 Subject: [Histonet] Thermo-Fisher Products In-Reply-To: <37A1F9CAB9E21C41B39F3653B620D13E093BA296@exchange2003.lc.local> References: <6DD7EFAF15E66C4583A82119CAF1590C0A449A3E@exch04.ahf2000.aultman.com> <37A1F9CAB9E21C41B39F3653B620D13E093BA296@exchange2003.lc.local> Message-ID: I have been a user of the Sakura Xpress 120 (2 units) for 5+ years and find them to be very reliable w/ excellent performance and they fit into the continuous workflow process exceptionally well. William DeSalvo B.S. HTL (ASCP) Production Manager, Sonora Quest Laboratories Chair Person, NSH Quality Control Committee wdesalvo.cac@hotmail.com > Date: Mon, 18 May 2009 10:41:07 -0400 > From: cbrya@lexclin.com > To: LMurphy2@aultman.com; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Thermo-Fisher Products > CC: > > We have an Excelsior that is about 5 years old. We have had a lot of > issues and have had service too many times in the past year to count. > We are looking at replacing it. Perhaps we got a lemon, but I don't > think I could recommend it based on our experience. Are there any other > processors anyone would like to recommend? Also, how about microwave > processors and IHC? > Thanks, > Carol > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leann M. > Murphy > Sent: Monday, May 11, 2009 10:33 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Thermo-Fisher Products > > I was just wondering what everyone's thoughts were on thermo products. > I have been demo-ing an Exlcelsior Processor and was wondering if > anyone had any problems. > > Let me know. > > > > LeAnn Murphy > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > NOTICE OF CONFIDENTIALITY > > This electronic message, including attachments, is for the sole > use of the named recipient and may contain confidential or > privileged information protected by the State of Kentucky and/or > Federal regulations. Any unauthorized review, use, disclosure, > copying or distribution is strictly prohibited. > > If you are not the intended recipient and have received this communication in error, please contact the sender and destroy > all copies of the original message. Thank you. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Hotmail? has a new way to see what's up with your friends. http://windowslive.com/Tutorial/Hotmail/WhatsNew?ocid=TXT_TAGLM_WL_HM_Tutorial_WhatsNew1_052009 From wdesalvo.cac <@t> hotmail.com Mon May 18 12:41:29 2009 From: wdesalvo.cac <@t> hotmail.com (WILLIAM DESALVO) Date: Mon May 18 12:41:33 2009 Subject: [Histonet] QA Manual Question In-Reply-To: <174282.82780.qm@web65712.mail.ac4.yahoo.com> References: <174282.82780.qm@web65712.mail.ac4.yahoo.com> Message-ID: What are you trying to track and what do you want to accomplish? Are you going to track and record defects/errors produced in your process, performance of the individuals, quality of the processes or all? This is a big project that will require setting up "controls" to monitor performance and it would be helpful to know what you now have in place. > Date: Fri, 15 May 2009 13:04:41 -0700 > From: arvidsonkristen@yahoo.com > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] QA Manual Question > > Hi All...Again. I guess I have a lot of questions lately. I have asked this before but I would like to ask it again. What are people doing for QA/QC? I have been asked by management to write a QA manual. I have some ideas but not really sure where to start. Thanks in advance for any input. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Hotmail? has a new way to see what's up with your friends. http://windowslive.com/Tutorial/Hotmail/WhatsNew?ocid=TXT_TAGLM_WL_HM_Tutorial_WhatsNew1_052009 From lpaveli1 <@t> hurleymc.com Mon May 18 12:45:11 2009 From: lpaveli1 <@t> hurleymc.com (Lynette Pavelich) Date: Mon May 18 12:45:24 2009 Subject: [Histonet] Alpha-naphthyl-butyrate Message-ID: <4A116667020000EE000293F8@smtp-gw.hurleymc.com> Check out the Sigma-Aldrich website at: www.sigmaaldrich.com 91A is part of a kit & 90A1 is the "select reagent packaged in gelatin capsules." Lynette Lynette Pavelich, HT(ASCP) Histology Supervisor Hurley Medical Center One Hurley Plaza Flint, MI 48503 ph: 810-257-9948 fax: 810-762-7082 >>> Stella Mireles 05/18/09 1:16 PM >>> Looking for the order number for Alpha-naphthyl-butyrate. I will be using it for the NSE procedure. I spoke with the Fisher 800 number and they were unable to help me. We use Fisher or Sigma products. Thanks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Mon May 18 13:46:32 2009 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Mon May 18 13:46:44 2009 Subject: AW: Re: AW: [Histonet] Chemicals that inactivate the primaryantibody In-Reply-To: <20090518181651.tx8fvdph6owks4cw@home.staff.uni-marburg.de> References: <261542.82019.qm@web65705.mail.ac4.yahoo.com><200905182337139539275@foxmail.com> <20090518181651.tx8fvdph6owks4cw@home.staff.uni-marburg.de> Message-ID: <527C3C2992B542F08F9403044824CDFB@dielangs.at> Hi Frauke, I've read the abcam protocol. They don't indicate what species the ab's = are from. Or I've overlooked it. What is the reason, why the second secondary ab doesn't bind to the = first primary ab?=20 Gudrun -----Urspr=A8=B9ngliche Nachricht----- Von: Dr. Frauke Neff [mailto:nefff@staff.uni-marburg.de]=20 Gesendet: Montag, 18. Mai 2009 18:17 An: tifei@foxmail.com Cc: Rene J Buesa; gu.lang; Merced MLeiker; histonet Betreff: Re: Re: AW: [Histonet] Chemicals that inactivate the primaryantibody Hi TF, we do IF with two 1.ABs out of mice using a sequential protocol from =20 abcam: www.abcam.com/technical We perform a second blocking step with serum (higher conc. than the =20 first one) after the first Fluorescence was added, do not perform =20 another retrieval procedure (my ABs need the same) and perform the =20 following incubation steps in the "dark" (we switch the light off and =20 cover the slides after adding the ABs) to prevent fading. Until now, I was sure, this procedure works but maybe the two antigens =20 don't colocalize but it is just a staining artifact ;-) Hope this helps, Frauke Zitat von TF : > Thanks all, Merced M Leiker is right: the double IHC procedure using =20 > two primary antibodies from same species. > > Some expert just sent me their procedure of Heat-interval based =20 > inactivation of the primary antibody for first antigen. > I am just seeking for a chemical way at room temperature. > > > 2009-05-18 > > > > TF > > > > =B7=A2=BC=FE=C8=CB=A3=BA Rene J Buesa > =B7=A2=CB=CD=CA=B1=BC=E4=A3=BA 2009-05-18 22:31:30 > =CA=D5=BC=FE=C8=CB=A3=BA tifei; gu.lang; Merced M Leiker > =B3=AD=CB=CD=A3=BA histonet > =D6=F7=CC=E2=A3=BA Re: AW: [Histonet] Chemicals that inactivate the = primary antibody > > I am completely confused. > IF you have an epitope in the tissue and you react with it an =20 > specific antibody "that is it". The epitope will have reacted and I =20 > do not see any way in which you can add another antibody to that =20 > initial epitope. From the reactive point of view, the epitope is =20 > "blocked" to react with another, unless the reaction is not totally =20 > specific and "there is room for another". > I just cannot imagine a way of doing this. > Ren=A8=A6 J. > > --- On Mon, 5/18/09, Merced M Leiker wrote: > > > From: Merced M Leiker > Subject: Re: AW: [Histonet] Chemicals that inactivate the primary = antibody > To: tifei@foxmail.com, gu.lang@gmx.at > Cc: histonet@lists.utsouthwestern.edu > Date: Monday, May 18, 2009, 9:42 AM > > > Hi TF and all interested, > > I think I know what you want, but unfortunately I don't know how to = answer > your question (it is something I'd like answered myself!!) To re-word = for > the sake of all interested: > > You want to perform double-immunofluorescent staining using 2 = primaries > that were raised in the same species. > > An additional question for clarification is: Do you want to do this on > paraffin or frozen sections? > > Maybe someone can help figure it out... > > Merced > > > --On Sunday, May 17, 2009 2:16 AM +0800 TF wrote: > >> But the heat also damage the fluorescnece... >> u need specific amplication kit anyway. >> >> >> 2009-05-17 >> >> >> >> TF >> >> >> >> =B7=A2=BC=FE=C8=CB=A3=BA Gudrun Lang >> =B7=A2=CB=CD=CA=B1=BC=E4=A3=BA 2009-05-17 02:00:18 >> =CA=D5=BC=FE=C8=CB=A3=BA tifei@foxmail.com >> =B3=AD=CB=CD=A3=BA histonet@lists.utsouthwestern.edu >> =D6=F7=CC=E2=A3=BA AW: [Histonet] Chemicals that inactivate the = primary antibody >> >> For doublestaining the primary undergoes denaturation through a = second >> HIER-step. The second secondary ab doesn't bind to the first primary. >> Therefore the binding sites must have been destroyed by heat in >> retrieval-buffer. >> Gudrun >> -----Urspr=FCngliche Nachricht----- >> Von: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von TF >> Gesendet: Samstag, 16. Mai 2009 18:48 >> An: Histonet@lists.utsouthwestern.edu >> Betreff: [Histonet] Chemicals that inactivate the primary antibody >> Hi all, just wonder what kind of treatments/chemicals can complete = block >> the binding of 2nd antibody to binded primary antibody on antigen? >> I tried HCl , but does not damage all the primaryantibody binding = sites - >> i can still see staining pattern finally. >> 2009-05-17 >> TF >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > Merced M Leiker > Cardiovascular Medicine > 348 Biomedical Research Building > State University of New York at Buffalo > 3435 Main St, Buffalo, NY 14214 > leiker@buffalo.edu > 716-829-6118 > > No trees were harmed in the sending of this email. > However, many electrons were severely inconvenienced. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From JMcCormick <@t> schosp.org Mon May 18 13:50:35 2009 From: JMcCormick <@t> schosp.org (McCormick, James) Date: Mon May 18 13:50:40 2009 Subject: [Histonet] coverslipping helper In-Reply-To: <232678.60898.qm@web65706.mail.ac4.yahoo.com> References: <232678.60898.qm@web65706.mail.ac4.yahoo.com> Message-ID: <0CA58D7DD405854899D129A03276B1330AAB0B9253@EXCHCCRMB.schosp.org> Rene, I think you're correct. In many of the old/antique supply catalogues there are listings for several types of "spring" coverslip clamping forceps that were used to "compress" the coverslip while drying the mountant, and to distribute Canada balsam evenly beneath the cover. When I used to make whole mounts of embryos etc and mount them in heavy balsam I used to place lead shot filled ampoules on the coverslips as a leveling weight while they dried. Regards, Jim, J.B.McCormick, M.D. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Monday, May 18, 2009 12:06 PM To: HISTONET; Andrea Grantham Subject: [Histonet] coverslipping helper Never heard of that, but perhaps she is referring to placing some small weight over the coverslipped section to help eliminate the bubbles by the weight, other than that, that is totally new for me. Ren? J. --- On Mon, 5/18/09, Andrea Grantham wrote: From: Andrea Grantham Subject: [Histonet] coverslipping helper To: "HISTONET" Date: Monday, May 18, 2009, 12:13 PM Good Monday morning! This may sound crazy...or not. I have to admit that I have never heard of this device before and I was wondering if any of you out in histoland could tell me what this is and where one might go to get it, if it were available for sale. Here goes: A student who was having a difficult time learning how to coverslip slides in my lab told me that she used to work for a doctor who invented a device to help coverslip. It was "like a magnet" and just eliminated all bubbles from under the coverslip. Anybody know? Andi Andrea Grantham, HT (ASCP) Senior Research Specialist University of Arizona Cell Biology and Anatomy Histology Service Laboratory P.O.Box 245044 Tucson, AZ 85724 algranth@email.arizona.edu Tel: 520.626.4415 Fax: 520.626.2097 "happy slicing and dicing and may all your stains work perfectly" - Paula Sicurello _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. 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From tifei <@t> foxmail.com Mon May 18 14:13:57 2009 From: tifei <@t> foxmail.com (TF) Date: Mon May 18 14:14:18 2009 Subject: AW: Re: AW: [Histonet] Chemicals that inactivate the primaryantibody References: <261542.82019.qm@web65705.mail.ac4.yahoo.com><200905182337139539275@foxmail.com>, <20090518181651.tx8fvdph6owks4cw@home.staff.uni-marburg.de> Message-ID: <200905190313522525992@foxmail.com> SSB0ZXN0ZWQgdGhlIHNlcXVlbnRpYWwgSUhDIHByb3RvY29sLi4udGhvdWdoIGEgYml0IGRpZmZl cmVudC4NCg0KSSBhbSB1c2luZyB0d28gbW91c2UgbW9ub2Nsb25hbCBhbnRpYm9keSwgYW5kIEdv YXQtYW50aSBNb3VzZSBBbGV4YSA0ODggLCA1NjgsIHJlc3BlY3RpdmVseS4NClRoZSAybmQgc2Vj b25kYXJ5IEFiIHdpbGwgYmluZCB0byB0aGUgMXN0IHByaW1hcnkgYW50aWJvZHksIGZvciBzdXJl LCB3aXRoIGFsbW9zdCA+ODAlIGJhY2tncm91bmQgKHlvdSBjYW4gdmlzdWFsaXplIHZlcnkgZGFy ayBjZWxsIG1vcnBob2xvZ3kgaW4gdGhlIG90aGVyIGNoYW5uZWwpLCBhbmQgYXJvdW5kIDEwLTUw JSBjbGVhciBjby1sYWJlbGluZyBvZiB0d28gZGlmZmVyZW50IGNlbGwgbWFya2Vycy4gV2l0aG91 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ZWx5IGluY29udmVuaWVuY2VkLg0KPg0KPg0KPiBfX19fX19fX19fX19fX19fX19fX19fX19fX19f X19fX19fX19fX19fX19fX19fXw0KPiBIaXN0b25ldCBtYWlsaW5nIGxpc3QNCj4gSGlzdG9uZXRA bGlzdHMudXRzb3V0aHdlc3Rlcm4uZWR1DQo+IGh0dHA6Ly9saXN0cy51dHNvdXRod2VzdGVybi5l ZHUvbWFpbG1hbi9saXN0aW5mby9oaXN0b25ldA0KPg0KX19fX19fX19fX19fX19fX19fX19fX19f X19fX19fX19fX19fX19fX19fX19fX18NCkhpc3RvbmV0IG1haWxpbmcgbGlzdA0KSGlzdG9uZXRA bGlzdHMudXRzb3V0aHdlc3Rlcm4uZWR1DQpodHRwOi8vbGlzdHMudXRzb3V0aHdlc3Rlcm4uZWR1 L21haWxtYW4vbGlzdGluZm8vaGlzdG9uZXQNCg== From rhbrown1 <@t> histocs.com Mon May 18 14:42:57 2009 From: rhbrown1 <@t> histocs.com (Leroy Brown) Date: Mon May 18 14:45:34 2009 Subject: [Histonet] looking for Larry Mahler Message-ID: Hi, Does anyone know how to contact Larry Mahler. Is or was with Scientific Instrument Service in TN. I am looking for some help with a IL MVP I tissue processor. I would like to also find anyone who might know about where to find parts for this older machine. LeRoy Brown HT(ASCP) HTL Everson, WA 98247 360-966-7300 www.histocs.com From CIngles <@t> uwhealth.org Mon May 18 15:30:04 2009 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Mon May 18 15:32:31 2009 Subject: [Histonet] Slide quality References: Message-ID: I think your techs have misplaced a digit. It should only be about 5-10% ammonium chloride. (plenty strong enough for ME thank you.) And no, we don't have IHC problems later. Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Saundra Johnson Sent: Fri 5/8/2009 12:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide quality Hi Bernice, I noticed a post in the histonet archives about using ammonia water to soak. Could I ask your opinion about how ammonia water could effect the IHC staining? I have come to work here and the two techs here soak in 50% ammonia water. That seems way to strong to me. What do you think? Saundra Johnson, HT(ASCP)QIHC Histology Supervisor NeoGenomics Laboratories "When Time Matters...And Results Count" 12701 Commonwealth Drive, Suite 9 Fort Myers, FL 33913 Phone: (239) 768-0600 Ext. 2229 Fax: (239) 768-0711 ________________________________ This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. E-mail transmission cannot be guaranteed to be secured or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The sender therefore does not accept liability for any errors or omissions in the contents of this message, which arise as a result of e-mail transmission. If verification is required please request a hard-copy version. NeoGenomics Laboratories, Suite 5, 12701 Commonwealth Dr, Fort Myers, FL 33913, http://www.neogenomics.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gentras <@t> vetmed.auburn.edu Mon May 18 15:57:54 2009 From: gentras <@t> vetmed.auburn.edu (Atoska Gentry) Date: Mon May 18 15:58:04 2009 Subject: [Histonet] Re: heartworm processing protocol Message-ID: <4A11CBD2.5090708@vetmed.auburn.edu> hello, will someone with a tried & proven protocol for processing heartworms for paraffin sectioning please share your protocol with me ASAP? My histonet archive search was unsuccessful. Thanks! Atoska From NMargaryan <@t> childrensmemorial.org Mon May 18 16:02:35 2009 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Mon May 18 16:04:31 2009 Subject: [Histonet] Fixation of Formalin vs Bouin's for IHC Message-ID: Hi histonetters, Please help. What is difference in procedure for IHC between 10%Formalin fixed tissue and in Bouin's fixed tissue? I used to use 10%FFPE tissue with Citrate buffer Antigen Retrieval for my IHC. Do I have to change my protocol for the Bouin's fixed tissue? Hope for you soon response and thanks in advance, Naira From mpence <@t> grhs.net Mon May 18 16:33:23 2009 From: mpence <@t> grhs.net (Mike Pence) Date: Mon May 18 16:33:32 2009 Subject: [Histonet] Special Stainers Message-ID: <661949901A768E4F9CC16D8AF8F2838C017A3B52@IS-E2K3.grhs.net> I am wanting to explore all my options. I am having some "real" issues with our automated special stainer and I am considering going to a new system. Can anyone tell me what systems are out on the market that preforms hands-free special stains. All options are open. Vendors welcome to reply. Mike Pence AP Supervisor Great River Medical Center From disbrc <@t> shands.ufl.edu Mon May 18 17:12:34 2009 From: disbrc <@t> shands.ufl.edu (Carrie Disbrow) Date: Mon May 18 17:12:42 2009 Subject: [Histonet] AFB Fites of cytospins Message-ID: <4A11A513.72AC.0059.0@shands.ufl.edu> Hello Histonet. Has anyone had experience and good results doing AFB Fites on cytospin slides from BAL's? Do you use the peanut oil/xylene (PO/Z) solution or just 100% peanut oil? Or does the cytospin slide need any peanut oil? After the deparaffinization step, if it can be called that, we are going to stain using a Dako Artisan special stainer. One problem I foresee is that the control will be de-arffinized in the traditional PO/Z while the patient slide may not. Also, I was told that Cytolyte contains alcohol and asked that the slides not be prepared using Cytolyte. Thanks for your comments. Please respond back to the histonet if possible and not my email address. Carrie From AnthonyH <@t> chw.edu.au Mon May 18 18:58:06 2009 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Mon May 18 18:58:17 2009 Subject: [Histonet] AFB Fites of cytospins In-Reply-To: <4A11A513.72AC.0059.0@shands.ufl.edu> Message-ID: If they are cytospins they will not need dewaxing so no vegetable oil. But having said this there is a school of thought that suggests that the use of vegetable oil may aid in the identification of Fite positive organisms (by ?coating the bugs). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Carrie Disbrow Sent: Tuesday, 19 May 2009 8:13 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] AFB Fites of cytospins Hello Histonet. Has anyone had experience and good results doing AFB Fites on cytospin slides from BAL's? Do you use the peanut oil/xylene (PO/Z) solution or just 100% peanut oil? Or does the cytospin slide need any peanut oil? After the deparaffinization step, if it can be called that, we are going to stain using a Dako Artisan special stainer. One problem I foresee is that the control will be de-arffinized in the traditional PO/Z while the patient slide may not. Also, I was told that Cytolyte contains alcohol and asked that the slides not be prepared using Cytolyte. Thanks for your comments. Please respond back to the histonet if possible and not my email address. Carrie _______________________________________________ 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 note 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 gachstetter <@t> yahoo.com Tue May 19 06:36:42 2009 From: gachstetter <@t> yahoo.com (Ginny .) Date: Tue May 19 06:36:45 2009 Subject: [Histonet] IHC cert. Message-ID: <111038.9791.qm@web51003.mail.re2.yahoo.com> Where can I find the study material needed to prepare for the IHC exam?? When on the ASCP website but found nothing.? From rjbuesa <@t> yahoo.com Tue May 19 07:28:57 2009 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue May 19 07:29:00 2009 Subject: [Histonet] AFB Fites of cytospins Message-ID: <392044.92619.qm@web65701.mail.ac4.yahoo.com> Carrie: The cytospin will not need dewaxing, as you point out, but I do not foresee any problem when compared with the control. You have to realize that dewaxing the control is intended only to?expose the tissue section to the reagents. As to the peanut oil if you are trying to stain for "common" Mycobacterium it is not necessary because their cover is quite strong. If you are trying to detect M. leprae it is when you use the mixture of peanut oil (or any other vegetable oil for that matter) mixed with xylene. The rationale is that the oil will "protect" the delicate cover of the organism. But again, not even for M. leprae you will need the mixture of oil and xylene if you are using a cytospin, because they do not require "deparafinization" (dewaxing). Ren? J. --- On Mon, 5/18/09, Carrie Disbrow wrote: From: Carrie Disbrow Subject: [Histonet] AFB Fites of cytospins To: histonet@lists.utsouthwestern.edu Date: Monday, May 18, 2009, 6:12 PM Hello Histonet. Has anyone had experience and good results doing AFB Fites on cytospin slides from BAL's? Do you use the peanut oil/xylene (PO/Z) solution or just 100% peanut oil? Or does the cytospin slide need any peanut oil? After the deparaffinization step, if it can be called that, we are going to stain using a Dako Artisan special stainer. One problem I foresee is that the control will be de-arffinized in the traditional PO/Z while the patient slide may not. Also, I was told that Cytolyte contains alcohol and asked that the slides not be prepared using Cytolyte. Thanks for your comments. Please respond back to the histonet if possible and not my email address. Carrie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Vickroy.Jim <@t> mhsil.com Tue May 19 08:24:52 2009 From: Vickroy.Jim <@t> mhsil.com (Vickroy, Jim) Date: Tue May 19 08:24:57 2009 Subject: [Histonet] Leica Peloris versus Microwave Processing Message-ID: <24A4826E8EF0964D86BC5317306F58A52BBB293EAB@mmc-mail.ad.mhsil.com> We are thinking about purchasing a new tissue processor. We have looked extensively into microwave tissue processors instead of the conventional tissue processors we are all used to. Recently two of my techs came back from a symposium and brought me literature on the Leica Peloris. The brochure makes some statements that it is faster than microwave processing. Has anyone used this processor and what has been your impression? Also do you have any opinion on how it matches up with microwave processors such as the Sakura Tissue Tek Express or the microwave processor by Milestone? Any input would be appreciated. Thanks Jim Vickroy BS, HT(ASCP) Technical Supervisor - Surgical and Autopsy Pathology Memorial Medical Center 217-788-4046 vickroy.jim@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. From cbrya <@t> lexclin.com Tue May 19 08:35:54 2009 From: cbrya <@t> lexclin.com (Carol Bryant) Date: Tue May 19 08:36:04 2009 Subject: [Histonet] Leica Peloris versus Microwave Processing In-Reply-To: <24A4826E8EF0964D86BC5317306F58A52BBB293EAB@mmc-mail.ad.mhsil.com> References: <24A4826E8EF0964D86BC5317306F58A52BBB293EAB@mmc-mail.ad.mhsil.com> Message-ID: <37A1F9CAB9E21C41B39F3653B620D13E093BA2AA@exchange2003.lc.local> Are there any issues with microwave processing and IHC? Thanks, Carol -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vickroy, Jim Sent: Tuesday, May 19, 2009 9:25 AM To: 'Histonet@lists.utsouthwestern.edu' Subject: [Histonet] Leica Peloris versus Microwave Processing We are thinking about purchasing a new tissue processor. We have looked extensively into microwave tissue processors instead of the conventional tissue processors we are all used to. Recently two of my techs came back from a symposium and brought me literature on the Leica Peloris. The brochure makes some statements that it is faster than microwave processing. Has anyone used this processor and what has been your impression? Also do you have any opinion on how it matches up with microwave processors such as the Sakura Tissue Tek Express or the microwave processor by Milestone? Any input would be appreciated. Thanks Jim Vickroy BS, HT(ASCP) Technical Supervisor - Surgical and Autopsy Pathology Memorial Medical Center 217-788-4046 vickroy.jim@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by the State of Kentucky and/or Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient and have received this communication in error, please contact the sender and destroy all copies of the original message. Thank you. From mpence <@t> grhs.net Tue May 19 08:47:23 2009 From: mpence <@t> grhs.net (Mike Pence) Date: Tue May 19 08:47:26 2009 Subject: [Histonet] Leica Peloris versus Microwave Processing In-Reply-To: <24A4826E8EF0964D86BC5317306F58A52BBB293EAB@mmc-mail.ad.mhsil.com> Message-ID: <661949901A768E4F9CC16D8AF8F2838C017A3B54@IS-E2K3.grhs.net> We are using the Peloris and love it! I process small bx type specimens in 1 hour throughout the day. I process 80% of the rest of our specimens in 4 hours and the large fatty specimens run for 9 hours. What I also like is that I do not have to deal with any of the microwave safety regs and er/pr hier2 issues with breast and other IHC problems with microwaving. (no I do not work for nor want to work for Leica). You may contact me off line to talk about in more detail. Mike AP Supervisor GRMC -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vickroy, Jim Sent: Tuesday, May 19, 2009 8:25 AM To: 'Histonet@lists.utsouthwestern.edu' Subject: [Histonet] Leica Peloris versus Microwave Processing We are thinking about purchasing a new tissue processor. We have looked extensively into microwave tissue processors instead of the conventional tissue processors we are all used to. Recently two of my techs came back from a symposium and brought me literature on the Leica Peloris. The brochure makes some statements that it is faster than microwave processing. Has anyone used this processor and what has been your impression? Also do you have any opinion on how it matches up with microwave processors such as the Sakura Tissue Tek Express or the microwave processor by Milestone? Any input would be appreciated. Thanks Jim Vickroy BS, HT(ASCP) Technical Supervisor - Surgical and Autopsy Pathology Memorial Medical Center 217-788-4046 vickroy.jim@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From relia1 <@t> earthlink.net Tue May 19 08:47:48 2009 From: relia1 <@t> earthlink.net (Pam Barker) Date: Tue May 19 08:47:54 2009 Subject: [Histonet] RELIA Histology Jobs Update Message-ID: Hi Histonetters! I hope everyone is having a great day. I wanted to do a quick post to tell you about some of the positions I am working on. All of my positions are full time permanent positions and my clients offer excellent compensation, benefits and relocation assistance. Here are the jobs I want to tell you about: Histology Supervisor - Springfield, MA Histology Supervisor - Los Angeles, CA Lead Histotechnologist (2nd Shift) North Shore of Boston Histotechnician/Histotechnologist - Los Angeles, CA Histotechnician/Histotechnologist - Night Shift NYC Histotechnician/Histotechnologist - Waco, TX If you or anyone you know might be interested in these opportunities or interested in having me look into positions in other areas of the country please contact me. I can be reached at 866-607-3542 or relia1@earthlink.net Thanks-Pam Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net www.myspace.com/pamatrelia www.twitter.com/pamatrelia From alyssa <@t> alliedsearchpartners.com Tue May 19 09:04:25 2009 From: alyssa <@t> alliedsearchpartners.com (Alyssa Peterson) Date: Tue May 19 09:04:31 2009 Subject: [Histonet] Job Opening In Long Island, NY Message-ID: ** *Hello Histonetters,* ** *I have an opening for a Lead/Supervisory Histotechnologist in Long Island, NY. * ** *Please review the job description and send an updated resume with any other questions to Alyssa@alliedsearchpartners.com * *If you are not interested but you know someone who is, we offer $1000 as a referral bonus if we place one of your referrals in a position. Please forward this email to anyone fit for this position. * *If this is not the job for you, but you are currently looking for a position, please send me a description of what you are looking for and your updated resume, as we have positions in your area!* Location: Long Island: *Shirly, NY* FULL TIME HISTOTECHNOLOGIST 3-5 years experience as a Histotechnologist, with at least 1-2 years in leadership role preferred and knowledge/ experience in immunohistochemistry is a plus. Benefit package includes moving expences, healthcare coverage, 401K with profit sharing plan,CME expences and life insurance. Come work with a growing private pathology laboratory in Suffolk County in Long Island, NY established in 1998. We work as a team in a friendly and professional environment to provide highest quality service to our client. We use state of art pathology equipments. We have excellent compensation package based on experience and productivity. -- Alyssa Peterson Allied Search Partners O: 770.621.2639 ext. 4 F: 770.621.2640 From donna <@t> milestonemed.com Tue May 19 08:54:49 2009 From: donna <@t> milestonemed.com (Donna Willis) Date: Tue May 19 09:08:10 2009 Subject: [Histonet] Leica Peloris versus Microwave Processing In-Reply-To: <37A1F9CAB9E21C41B39F3653B620D13E093BA2AA@exchange2003.lc.local> References: <24A4826E8EF0964D86BC5317306F58A52BBB293EAB@mmc-mail.ad.mhsil.com><37A1F9CAB9E21C41B39F3653B620D13E093BA2AA@exchange2003.lc.local> Message-ID: <908253790-1242741353-cardhu_decombobulator_blackberry.rim.net-500061931-@bxe1085.bisx.prod.on.blackberry> No issues using the Milestone products. Donna Sent via BlackBerry by AT&T -----Original Message----- From: "Carol Bryant" Date: Tue, 19 May 2009 09:35:54 To: Vickroy, Jim; Subject: RE: [Histonet] Leica Peloris versus Microwave Processing Are there any issues with microwave processing and IHC? Thanks, Carol -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vickroy, Jim Sent: Tuesday, May 19, 2009 9:25 AM To: 'Histonet@lists.utsouthwestern.edu' Subject: [Histonet] Leica Peloris versus Microwave Processing We are thinking about purchasing a new tissue processor. We have looked extensively into microwave tissue processors instead of the conventional tissue processors we are all used to. Recently two of my techs came back from a symposium and brought me literature on the Leica Peloris. The brochure makes some statements that it is faster than microwave processing. Has anyone used this processor and what has been your impression? Also do you have any opinion on how it matches up with microwave processors such as the Sakura Tissue Tek Express or the microwave processor by Milestone? Any input would be appreciated. Thanks Jim Vickroy BS, HT(ASCP) Technical Supervisor - Surgical and Autopsy Pathology Memorial Medical Center 217-788-4046 vickroy.jim@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by the State of Kentucky and/or Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient and have received this communication in error, please contact the sender 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 anitaibsc <@t> aol.com Tue May 19 09:10:51 2009 From: anitaibsc <@t> aol.com (anitaibsc@aol.com) Date: Tue May 19 09:11:15 2009 Subject: [Histonet] Autofluorescence Message-ID: <8CBA6A2C0698F03-F1C-2393@webmail-dd06.sysops.aol.com> Hello Netters, We are trying to do do immunofluorescence on FFPE tissue sections; we always get autofluorescence. We have tried Sodium Borohydride treatment, does not work very well. Does anyone have good method to remove this autofluorescence from FFPE tissue sections. We have never tried Toluidine blue, Fe2/Fe3, Phosphomolbidic acid. Thanks Bader Siddiki, PhD www.ImmunoBioScience.com From jqb7 <@t> cdc.gov Tue May 19 09:54:11 2009 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine (CDC/CCID/NCZVED)) Date: Tue May 19 09:54:37 2009 Subject: [Histonet] Leica Peloris versus Microwave Processing In-Reply-To: <908253790-1242741353-cardhu_decombobulator_blackberry.rim.net-500061931-@bxe1085.bisx.prod.on.blackberry> References: <24A4826E8EF0964D86BC5317306F58A52BBB293EAB@mmc-mail.ad.mhsil.com><37A1F9CAB9E21C41B39F3653B620D13E093BA2AA@exchange2003.lc.local> <908253790-1242741353-cardhu_decombobulator_blackberry.rim.net-500061931-@bxe1085.bisx.prod.on.blackberry> Message-ID: <9A16CB5D55FC1648ADF11B63E72A1BE101AD3185@LTA3VS011.ees.hhs.gov> We use the Sakura Xpress with no problems. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Donna Willis Sent: Tuesday, May 19, 2009 9:55 AM To: Carol Bryant; histonet-bounces@lists.utsouthwestern.edu; Mr. Jim Vickroy; Histonet Cc: Dave Sanford; Jes Strong Subject: Re: [Histonet] Leica Peloris versus Microwave Processing No issues using the Milestone products. Donna Sent via BlackBerry by AT&T -----Original Message----- From: "Carol Bryant" Date: Tue, 19 May 2009 09:35:54 To: Vickroy, Jim; Subject: RE: [Histonet] Leica Peloris versus Microwave Processing Are there any issues with microwave processing and IHC? Thanks, Carol -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vickroy, Jim Sent: Tuesday, May 19, 2009 9:25 AM To: 'Histonet@lists.utsouthwestern.edu' Subject: [Histonet] Leica Peloris versus Microwave Processing We are thinking about purchasing a new tissue processor. We have looked extensively into microwave tissue processors instead of the conventional tissue processors we are all used to. Recently two of my techs came back from a symposium and brought me literature on the Leica Peloris. The brochure makes some statements that it is faster than microwave processing. Has anyone used this processor and what has been your impression? Also do you have any opinion on how it matches up with microwave processors such as the Sakura Tissue Tek Express or the microwave processor by Milestone? Any input would be appreciated. Thanks Jim Vickroy BS, HT(ASCP) Technical Supervisor - Surgical and Autopsy Pathology Memorial Medical Center 217-788-4046 vickroy.jim@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by the State of Kentucky and/or Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient and have received this communication in error, please contact the sender and destroy all copies of the original message. 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 Kathleen.Cormier <@t> crl.com Tue May 19 10:02:07 2009 From: Kathleen.Cormier <@t> crl.com (Cormier, Kathleen) Date: Tue May 19 10:02:18 2009 Subject: [Histonet] ? potassium permangante filters, end point Message-ID: <7862ACBA8359E04FA9B0C5375A2E6F4644F935@shr-sv0014.na01.crl.com> Hello Netters, I was wondering if anyone has a reference or knows where I can track down information on determining the end point of potassium permanganate filters by dropping the used portion of the filter particulate into water. I have heard that if you drop the used filter particulate in to water and it turns a certain color (purple or brown) you can find out if the filter is used up. Rumor? Outright lie? Correct? (and the manufacturer at this point is useless on determining the real end point of the filters for our HVAC system.) Thanks for any help! Kathy . From rgrow <@t> bmnet.com Tue May 19 10:24:54 2009 From: rgrow <@t> bmnet.com (rgrow@bmnet.com) Date: Tue May 19 10:24:56 2009 Subject: [Histonet] Job opening in East Tennessee Message-ID: Blount Memorial Hospital in Maryville, TN has a histology technician position open Monday thru Friday 7-3:30. We are located just minutes from the beautiful Smoky Mountains National Park and experience 4 wonderful seasons! All types of outdoor activities are possible. Maryville is host to the annual Foothills Fall Festival with top name entertainment and crafts, and just 30 minutes from Knoxville's culture events and entertainment, as well as UT football, basketball, etc. Blount Memorial has an attractive benefits package and possible relocation assistance for the right candidate. Applicants must meet the educational and training requirements necessary for certification by the American Society of Clinical Pathology as a Histology Technician or have experience equal to certification. General histology experience preferred. Must demonstrate competency and successfully complete the on-the-job orientation through the histology section of the laboratory. Performs all duties of a Histology Technician and other duties as assigned. Technically, it is currently listed as part time, but there will be 40 hrs/wk needed, and will become a full time position. Anyone interested please visit our website at blountmemorial.org. to fill out an application and attach a resume. Or, you may just send me a resume. Thanks, Renee Grow, BA., HT (ASCP) rgrow@bmnet.com Histology Supervisor Blount Memorial Hospital 907 E. Lamar Alexander Pkwy. Maryville, TN 37804-5016 (865) 977-4744 (865) 977-5766 Fax ________________________________ This communication may contain protected health information (PHI) that is legally protected from inappropriate disclosure by the Privacy Standards of the Health Insurance Portability and Accountability Act (HIPAA) and relevant Tennessee Laws. If you are not the intended recipient, please note that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this message in error, you should notify the sender immediately by telephone or by return e-mail and delete this message from your computer. Direct questions to the Blount Memorial Hospital Privacy Officer at 865-977-4675. From gayle.callis <@t> bresnan.net Tue May 19 10:33:40 2009 From: gayle.callis <@t> bresnan.net (gayle callis) Date: Tue May 19 10:34:00 2009 Subject: [Histonet] RE: Autofluorescence with FFPE tissue Message-ID: <000a01c9d897$30ca10a0$925e31e0$@callis@bresnan.net> You wrote: We are trying to do do immunofluorescence on FFPE tissue sections; we always get autofluorescence. We have tried Sodium Borohydride treatment, does not work very well. Does anyone have good method to remove this autofluorescence from FFPE tissue sections. We have never tried Toluidine blue, Fe2/Fe3, Phosphomolbidic acid. Thanks Bader Siddiki, PhD www.ImmunoBioScience.com ********************************************** Bader, You may not be able to completely get rid of aldehyde induced autofluorescence. Another thing to try is 100 mM glycine in pH 7.4 buffer, can be TRIS buffer or PBS. After deparaffinizing and rehydrating into water, place sections in this solution for 15 to 20 minutes, rinse and proceed with staining. One can also soak the tissues prior to processing in the same solution for an hour or so. We have not tried the latter, but working with sections has been successful with minimal, no more than 8 hour fixation in NBF. Minimal fixation may not be ideal since it may result in incomplete fixation for your tissues. If autofluorescence persists, you can use a Near Infrared Fluorophore to eliminate autofluorescence that way OR use the autofluorescence as a "Counterstain" by using an extremely bright red fluorophore that resists photobleaching e.g. Alexa 594 from Molecular Probes. Go to http://www.uhnres.utoronto.ca/facilities/wcif/fdownload2.html You can download a wonderful discussion on autofluorescence: causes and cures. This document has more ways of eliminating autofluorescence or links for doing so. In this document, it tells how to do the sodium borohydride treatment from a protocol written by Kramer along with Ian Clements (Molecular Probes). The solution works best when applied to the section while the solutions it still "fizzing" and for PFA fixed tissue (very similar to NBF fixed tissues), the solution is applied 3 times for 10 minutes each time to maintain freshness of the sodium borohydride. There was a recent discussion on Histonet on how to dispose of this chemical. OR you can use fresh tissue frozen sections fixed with cold acetone - as we do. We do not have autofluorescence problems when we do this. I have a synopsis of methods put on Histonet some time ago, and will be happy to forward that to you as an attachment. Good luck Gayle Callis HTL(ASCP)HT,MT Bozeman MT 59715 From pruegg <@t> ihctech.net Tue May 19 10:47:55 2009 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Tue May 19 10:47:55 2009 Subject: [Histonet] HISTOLOGY, COVERSLIPPER; Microm CTM-6 Glass Coverslipper Cat. # 970010 Message-ID: Does anyone in histoland use this coverslipper? What do you like about it? What do you not like about it? Best regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From gayle.callis <@t> bresnan.net Tue May 19 11:18:47 2009 From: gayle.callis <@t> bresnan.net (gayle callis) Date: Tue May 19 11:19:06 2009 Subject: [Histonet] Working with same host species primaries for double staining Message-ID: <000301c9d89d$7e00da60$7a028f20$@callis@bresnan.net> We do sequential immunostaining to avoid cross reaction of a secondary antibody to the next primary antibody, and also perform a normal serum block that binds to the first secondary antibody: Two examples using two rat antimouse primaries and with immunofluorescent method. This is done on fresh tissue frozen sections, so no NBF or PFA is involved, eliminating the need for antigen retrieval. First normal serum block is matched to host of first secondary primary e.g. donkey serum First primary is rat antimouse First secondary is donkey anti rat (F(ab')2 frag of IgG-fluorophore conjugate SPECIAL NORMAL RAT SERUM BLOCK, 10% FOR 30 MINUTES. This will bind to the secondary antibody just used and eliminate a possible binding of this secondary to the next primary antibody. Rinse very well. Second primary is rat antiMouse #2 secondary antibody is can be a goat antiRat- fluorophore conjugate Rinse very well. Our favorite method that works best for us, totally eliminating a secondary antibody in the last sequence is: Normal serum block matched to secondary antibody Streptavidin biotin kit block First primary is rat antimouse First secondary is donkey anti rat (F(ab')2 frag of IgG-fluorophore conjugate 10% Normal rat serum block for 30 minutes as done above Second primary antibody-biotinylated Streptavidin-Alexa fluor dye The best double staining method for enzyme immunohistochemistry where the tissue is FFPE and needs two antigen retrieval is to use Chris van der Loos method. He published this in J of Histotechnology, September, 2008 and has also presented a workshop using this method. He is an expert on performing double and even triple immunostaining and will be happy to discuss your protocol with you and also send you a copy of his publication. c.m.vanderloos@amc.uva.nl Good luck Gayle Callis HTL(ASCP)HT,MT Bozeman MT From mwich <@t> 7thwavelabs.com Tue May 19 11:33:50 2009 From: mwich <@t> 7thwavelabs.com (Michele Wich) Date: Tue May 19 11:33:57 2009 Subject: [Histonet] MetAP-2 Message-ID: <62A8156F8071C8439080D626DF8C33A65D8C3A@wave-mail.7thwave.local> Has anyone successfully used Santa Cruz's MetAP-2 for IHC in FFPE mouse and rat sections? I am starting for scratch and am trying to find info on dilution range and antigen retrieval methods. The spec sheet only lists recommendations for applications other than IHC. Any info is greatly appreciated! This communication is intended solely for the use of the addressee and may contain information that is legally privileged, confidential or exempt from disclosure. If you are not the intended recipient, please note that any dissemination, distribution, or copying of this communication is strictly prohibited. Anyone who receives this message in error should notify the sender immediately and delete it from his or her computer From cmnorman <@t> aacc.edu Mon May 18 16:00:17 2009 From: cmnorman <@t> aacc.edu (Norman, Michael) Date: Tue May 19 12:09:00 2009 Subject: [Histonet] Re Nikon Diaphot 300 Message-ID: <3614638D4C701B438F335F0EFA8F6EBC68C6271112@AACC-MAILBOX.aacc.cc.md.us> Does anyone have a .pdf copy of the instructions manual for the Nikon Diaphot 300 Inverted Microscope they could send me? Thanks, Michael Norman ________________________________ The information contained in this email may be confidential and/or legally privileged. It has been sent for the sole use of the intended recipient(s). If the reader of this message is not an intended recipient, you are hereby notified that any unauthorized review, use, disclosure, dissemination, distribution, or copying of this communication, or any of its content, is strictly prohibited. If you have received this communication in error, please contact the sender by reply email and destroy all copies of the original message. Thank you. From Virginia.Achstetter <@t> us.army.mil Tue May 19 06:31:58 2009 From: Virginia.Achstetter <@t> us.army.mil (Achstetter, Virginia A.) Date: Tue May 19 12:09:02 2009 Subject: [Histonet] IHC certification Message-ID: <127D612DBCD77546A12C4746F7491C7F053C1AD6@lewis.afip.osd.mil> How do I go about finding the study material needed for the IHC exam, how do I prepare, etc. Thanks Ginny Achstetter From JimR0712 <@t> comcast.net Tue May 19 12:14:08 2009 From: JimR0712 <@t> comcast.net (JimR0712@comcast.net) Date: Tue May 19 12:14:14 2009 Subject: [Histonet] Career Opportunity in Chicago area Message-ID: <387056283.11337581242753248724.JavaMail.root@sz0098a.emeryville.ca.mail.comcast.net> Last week, my former employer, Loyola University Medical Center in Maywood, Il., layed off 440 employees (about 8 % of the workforce). I was the Manager of Anatomic Pathology (Histo/Cyto/Autopsy/Transcription/MohsLabs/Clerical Support) and now am looking for a career opportunity in the Chicago area. Please let me know if you hear of any career opportunities. Thanks. From JimR0712 <@t> comcast.net Tue May 19 12:47:43 2009 From: JimR0712 <@t> comcast.net (JimR0712@comcast.net) Date: Tue May 19 12:47:46 2009 Subject: [Histonet] Looking for career opportunity in Chicago area In-Reply-To: <387056283.11337581242753248724.JavaMail.root@sz0098a.emeryville.ca.mail.comcast.net> Message-ID: <1482254971.11355411242755263324.JavaMail.root@sz0098a.emeryville.ca.mail.comcast.net> I am looking for a career opportunity. ----- Forwarded Message ----- From: JimR0712@comcast.net To: "Histonet2" Sent: Tuesday, May 19, 2009 12:14:08 PM GMT -06:00 US/Canada Central Subject: Career Opportunity in Chicago area Last week, my former employer, Loyola University Medical Center in Maywood, Il., layed off 440 employees (about 8 % of the workforce). I was the Manager of Anatomic Pathology (Histo/Cyto/Autopsy/Transcription/MohsLabs/Clerical Support) and now am looking for a career opportunity in the Chicago area. Please let me know if you hear of any career opportunities. Thanks. From af46 <@t> buffalo.edu Tue May 19 13:39:52 2009 From: af46 <@t> buffalo.edu (af46@buffalo.edu) Date: Tue May 19 13:39:57 2009 Subject: [Histonet] Trying to post Message-ID: <14581.1242758392@buffalo.edu> just testing to see if I can post From jdhisto <@t> yahoo.com Tue May 19 14:45:13 2009 From: jdhisto <@t> yahoo.com (Jonathan De La Rosa) Date: Tue May 19 14:45:18 2009 Subject: [Histonet] CLIA regulations Message-ID: <638245.37316.qm@web110701.mail.gq1.yahoo.com> Hello Histoneters, I was wondering if someone could?contact me?offline about CLIA regulations and/or inspections. I have a few concerns. Any help would be appreciated. Thanks in advance. JD jdhisto@yahoo.com From LBrown <@t> getvitalized.com Tue May 19 15:08:33 2009 From: LBrown <@t> getvitalized.com (Brown, Lori) Date: Tue May 19 15:08:37 2009 Subject: [Histonet] PathNet HLA Module Message-ID: I'm hoping to network with some wonderful Pathologists (and informaticists). I'm wondering if anyone out there has ever used or been a part of an implementation of Cerner Millennium's PathNet HLA module for tissue transplant. Any networking and advice is greatly appreciated!! Lori M. Brown, SPHR, CIR Recruiting Manager Vitalize Consulting Solutions | 410-392-5332 o | 484-356-3781 c | 484-734-6168 f | www.getvitalized.com ________________________________ Erase your problems. See how in booth #530 at the International MUSE May 25th - 29th. ________________________________ NOTICE: This message and any included attachments are from VITALIZE CONSULTING SOLUTIONS, INC. and are intended only for the addressee(s). The information contained herein may include intellectual property or privileged or otherwise confidential information. If you are not the named recipient, you are prohibited to forward, print, copy, distribute, disclose, or use such information and your actions may be unlawful. If an addressing or transmission error has misdirected the email, or you have reason to believe you are not authorized to receive it, please promptly delete this message and notify the sender by e-mail with a copy to vcs@getvitalized.com. From disbrc <@t> shands.ufl.edu Tue May 19 16:19:36 2009 From: disbrc <@t> shands.ufl.edu (Carrie Disbrow) Date: Tue May 19 16:19:55 2009 Subject: [Histonet] PO/Zylene on cytospins for Fites Message-ID: <4A12EA2D.72AC.0059.0@shands.ufl.edu> Hi Rene, Thanks for the info. We are doing a AFB Fites for Nocardia. There is filamentous bacteria in the patient GMS and the docs are differentiating the type. I did not deparaffinize the patient cytospin slides and I did deparaffinize the patient control with Z/PO. The problem I had been anticipating was that the patient and control did not have the same procedure. I was thinking of it as a CAP issue,,,, but then realized we don't have the same preparation for any stains when using cytospins and controls, ie GMS, Mucin, irons, all have controls that need deparaffinization. DUH! Anyway the stain did not work. It looks like the Methylene blue is too heavy and blocking the carbol fuschin. We use Dako's Artisan stainer, which I love and are having to rework this stain. Thanks again, Carrie From disbrc <@t> shands.ufl.edu Tue May 19 16:22:45 2009 From: disbrc <@t> shands.ufl.edu (Carrie Disbrow) Date: Tue May 19 16:23:01 2009 Subject: [Histonet] Hi Tony, Message-ID: <4A12EAEA.72AC.0059.0@shands.ufl.edu> Hi Tony, Thanks for the info. I understand the discussion topic. I just wish their was one straight answer! But then this is histology! Carrie From rhbrown1 <@t> histocs.com Tue May 19 16:33:22 2009 From: rhbrown1 <@t> histocs.com (Leroy Brown) Date: Tue May 19 16:36:12 2009 Subject: [Histonet] Thanks to those who helped find Larry M. plus Message-ID: I was able to speak to Larry M. about my tissue processor problems. thank you all for your help. I am now looking for perhaps a broken MVP-1 or 2 since I am no longer able to get the CRT display parts to fix mine. they are interchangeable and if I could find a discarded MVP I could get mine running again. Anyone have a mothballed one sitting in the basement? Thanks LeRoy Brown HT(ASCP)HTL www.histocs.com Everson, Wa 98247 From baza0013 <@t> umn.edu Tue May 19 16:44:08 2009 From: baza0013 <@t> umn.edu (Adam Bazama) Date: Tue May 19 16:44:25 2009 Subject: [Histonet] Cryoprotection Issues with Unfixed Mouse Hearts Message-ID: <000001c9d8ca$f0d14610$d273d230$@edu> Does anyone have any suggestions or protocols for cryoprotecting unfixed mouse heart tissue? Some of the ones I have been cryosectioning lately have been damaged, I think because of improper cryoprotection. Thanks! Adam baza0013@umn.edu From gayle.callis <@t> bresnan.net Tue May 19 17:49:36 2009 From: gayle.callis <@t> bresnan.net (gayle callis) Date: Tue May 19 17:49:56 2009 Subject: [Histonet] Cryoprotection Issues with Unfixed Mouse Hearts In-Reply-To: <000001c9d8ca$f0d14610$d273d230$@edu> References: <000001c9d8ca$f0d14610$d273d230$@edu> Message-ID: <002101c9d8d4$17177750$454665f0$@callis@bresnan.net> Adam, It is not a good to sucrose cryoprotect fresh tissues. Only well fixed tissues should be cryoprotected to lessen freezing artifact caused by large water ice crystals. Charles Scouten, on his website discusses this, and if one cryoprotects fresh tissues, you change the osmolarity within tissues/cells and the tissues shrivel up. This is probably the damage/changes you are observing. Tissues that are well fixed with NBF or a PFA mixture should be sucrose cryoprotected. For fresh tissues, dissect heart from mouse, embed and snap freeze with a proper RAPIC snap freezing method. If you would like, I will privately send a simple snap freezing that eliminates use of precooled isopentane. Avoid freezing in a cryostat or a freezer, even -80C as freezing will be too slow and at warmer temperture than true/rapidsnap freezing methods. This will also cause freezing artifact in both fresh or sucrose cryoprotected fixed tissues. Be sure to read Dr. Scouten's comments on snap freezing and its effects on tissues - it is very informative. Gayle Callis HTL(ASCP)HT,MT Bozeman MT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Adam Bazama Sent: Tuesday, May 19, 2009 3:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cryoprotection Issues with Unfixed Mouse Hearts Does anyone have any suggestions or protocols for cryoprotecting unfixed mouse heart tissue? Some of the ones I have been cryosectioning lately have been damaged, I think because of improper cryoprotection. Thanks! Adam baza0013@umn.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Tue May 19 19:19:46 2009 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Tue May 19 19:19:52 2009 Subject: [Histonet] IHC cert. In-Reply-To: <111038.9791.qm@web51003.mail.re2.yahoo.com> Message-ID: For all information about certification exams (e.g., HT, HTL, SLS (specialist in laboratory safety), or on qualification exams (e.g., QIHC), go to www.ascp.org Click on Laboratory Professional Click on Certification Click on either "Get Certified" or "Get Qualified" Under Get Qualified, there is information on study material (#5) - Reading list - Topic outline Peggy A. Wenk, HTL(ASCP)SLS Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ginny . Sent: Tuesday, May 19, 2009 7:37 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC cert. Where can I find the study material needed to prepare for the IHC exam?? When on the ASCP website but found nothing.? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jamie.erickson <@t> abbott.com Wed May 20 08:32:09 2009 From: jamie.erickson <@t> abbott.com (Jamie E Erickson) Date: Wed May 20 08:32:16 2009 Subject: [Histonet] Sucrose infiltration and Bone question... Message-ID: Hi All, I have two question about sucrose infiltration....First, I have infiltrated many mouse brain with sucrose 30% in the past and it has always been at 4degrees C (dc) and I'm asking myself now why? My mouse brain are fixed in paraformaldehyde or formaldehyde for 24 hours prior so why sucrose @ 4dc? Would it not infiltrate better (faster) at room temp? I could not find out why 4dc... My real problem is that I am tiring to get good mouse tibia/femur frozen sections and I am using D-profile blade and a tape transfer method. It worked great on my rats sections but not mouse so I think the sucrose is not getting in to the bone. I can't use the sinking tissue as an indicator because the bone sinks right away. Should I go longer in sucrose (maybe 48 Hrs) but at 4dc or room temp ? Would adding 2 % DSMO to sucrose facilitate penetration..?? Any suggestions would be great.. Thanks have a great day.. Jamie _______________________________ Jamie Erickson Sr. Research Associate II M.S. HTL (ASCP) Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com From Annette.Fletcher <@t> providence.org Tue May 19 13:23:22 2009 From: Annette.Fletcher <@t> providence.org (Fletcher, Annette M) Date: Wed May 20 09:22:40 2009 Subject: [Histonet] Histotechnician 2 opportunity in Portland, OR Message-ID: <284596E350CB0743BB500B18475E7A0B022A6562@wn1223.or.providence.org> Good Afternoon Histonetters, Annette Fletcher here with Providence Health and Services in Portland, Oregon. We are seeking a Histotech for our Regional Lab in Portland, OR. This is an opportunity to work for a visionary Manager who is dedicated to growing and developing her Histo staff. We have a state of the art lab, highly competitive compensation and an excellent benefit plan. We also offer relocation assistance. You can email me at annette.fletcher@providence.org with your interest. Thank you, Annette M Fletcher Senior Recruiter Providence Health & Services, Employment 1235 NE 47th Avenue - Suite 200 Portland, OR 97213 T: 503-215-5840 F: 503-215-5770 Annette.fletcher@providence.org Confidentiality notice: The information contained in this email message including attachments is confidential and is intended only for the use of the individual or entity named above and others who have been specifically authorized to receive it. If you are not the intended recipient, you are hereby notified that any use, unauthorized dissemination, distribution, or copying of this communication is strictly prohibited. If you have received this communication in error, please delete immediately or if any problems occur with transmission, please notify me immediately by telephone at 503-215-5840 or by reply e-mail at annette.fletcher@providence.org. Thank you! DISCLAIMER: This message is intended for the sole use of the addressee, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If you are not the addressee you are hereby notified that you may not use, copy, disclose, or distribute to anyone the message or any information contained in the message. If you have received this message in error, please immediately advise the sender by reply email and delete this message. From j_mcanally <@t> hotmail.com Wed May 20 09:32:11 2009 From: j_mcanally <@t> hotmail.com (Jennifer McAnally) Date: Wed May 20 09:32:11 2009 Subject: [Histonet] Job Opening In Sacramento, CA Message-ID: ** *Hello Histonetters,* ** *I have an opening for a Lead/Supervisory Histotechnologist in Sacramento, CA. * ** * Please review the job description below. If you know someone qualified for this position, please forward this information. For anyone interested, please forward your resume to jennifer@phcconsulting.com for consideration. This Lab is part of a multi-lab company. This particular Lab processes around 300 cases a day. This person will be accountable for the direct supervision and operation of the lab and staff. We need someone who can support the company's growth oriented business plan and has an entrepreneurial spirit. This person will be vital to the overall success of the lab so the following characteristics are necessary: Great leadership skills, Ability to train others, Able to smoothly manage the workflow of the lab, Great attitude, energetic and pleasant personality. 3-5 years experience as a Histotechnologist, with at least 1-2 years in leadership role preferred. Salary range is $70k to $95k depending on experience. Performance bonuses, full benefits, healthcare, health savings accounts, 401k and accrued vacation. Relo package offered for the right person. Jennifer McAnally PHC Consulting 888-263-5688 ext 300 From ktuttle <@t> umm.edu Wed May 20 10:26:32 2009 From: ktuttle <@t> umm.edu (Kimberly Tuttle) Date: Wed May 20 10:27:07 2009 Subject: [Histonet] microwave antigen retrieval Message-ID: <4A13E8E7.90CE.001A.3@umm.edu> Can someone post instructions for microwave antigen retrieval... my pressure cooker broke and I need an immediate solution. Kimberly C. Tuttle HT (ASCP) Pathology Biorepository and Research Core University of Maryland Room NBW58, UMMC 22 S. Greene St Baltimore, MD 21201 (410) 328-5524 (410) 328-5508 fax This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. From akemiat3377 <@t> yahoo.com Wed May 20 10:34:20 2009 From: akemiat3377 <@t> yahoo.com (Akemi Allison-Tacha) Date: Wed May 20 10:34:23 2009 Subject: [Histonet] (Tissue Processor's) Used, Refurbished or Lease Agreement Message-ID: <780411.19431.qm@web31301.mail.mud.yahoo.com> Good Morning out there in Histo Land! Calling all Vendors. ?I have recently accepted a new position as the Histology Manager at APMG Labs in Los Gatos, CA. ?We are down one Tissue Processor. ?The Executive Administrator thought we would be purchasing a used VIP from one of our affiliates, but unfortunately this possibility has fallen through. ? I would appreciate any Vendors who have Tissue Processor's for sale that are Used or Refurbished. ?I am also open to the possibility of?negotiating?lease agreements. ?I realize warranties and service agreements are extremely important. Please e-mail me any available options. ? Thanks,Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Ct. Los Gatos, CA 95032 W: (408)884.2718? W: E-Mail: aallison-tacha@apmglab.com P: E-Mail: akemiat3377@yahoo.com From rjbuesa <@t> yahoo.com Wed May 20 11:05:18 2009 From: rjbuesa <@t> yahoo.com (rjbuesa@yahoo.com) Date: Wed May 20 11:05:22 2009 Subject: [Histonet] microwave antigen retrieval Message-ID: <848945.6183.qm@web65710.mail.ac4.yahoo.com> You use the MW to heat the HIER solution and the slides in it. Once the buffer boiled with the slides, remove the container with the slides and place it in either a water bath or a steamer at 98?C during 20 minutes. After that place the container over the counter during another 20 minutes. Wash with PBS and start the staining procedure (either manual or automated). Ren? J. --- On Wed, 5/20/09, Kimberly Tuttle wrote: From: Kimberly Tuttle Subject: [Histonet] microwave antigen retrieval To: "histonet" Date: Wednesday, May 20, 2009, 11:26 AM Can someone post instructions for microwave antigen retrieval... my pressure cooker broke and I need an immediate solution. Kimberly C. Tuttle? HT (ASCP) Pathology Biorepository and Research Core University of Maryland Room NBW58, UMMC 22 S. Greene St Baltimore, MD 21201 (410) 328-5524 (410) 328-5508 fax This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gayle.callis <@t> bresnan.net Wed May 20 12:14:32 2009 From: gayle.callis <@t> bresnan.net (gayle callis) Date: Wed May 20 12:14:54 2009 Subject: [Histonet] Sucrose infiltration and Bone question... In-Reply-To: References: Message-ID: <001b01c9d96e$7258ec30$570ac490$@callis@bresnan.net> Jamie, One thing to try is put your mouse bones under vacuum when you cryoprotect them. Vacuum will speed up the exchange and when you don't see bubbles coming off bone anymore it should be infiltrated - hopefully. I have always wondered about using 4C temps and just did it without asking questions. However, I have seen replies on Histonet where RT was used. I wonder if the rationale for 4C is that the sucrose could get some type of unwanted growing contaminant over a longer time period. If you are not doing IHC with chromogens, you could put sodium azide preservative in the sucrose to counteract the growth issues. Try suspending the bone in the sucrose so the solution surrounds the bones totally. If you have a vacuum oven at ambient RT it will even nicer so you don't have to deal with greasing a vacuum dessicator. We use O Ring Wheaton vacuum dessicators that do not need to be greased and easy to clean. I suggest you access this publication and as a member of NSH, it will be free to you if you do not have the journal on hand. John Tarpley, Preparation and sectioning of undecalcified frozen rodent long bones and joints using a tape transfer system. J Histotechnology 26(4):41, 2003. John worked with rat bones but his method was very successful. His preparation protocol was quite detailed with some interesting special ways of working with bones. If worked for him on rat bones, it may help you with the tiny difficult mouse bones. Try contacting John Baker to see if he can help you too. Info is found below. I think he dealt with mouse bone and the Cryojane. bakerj@umich.edu (john baker) John A. Baker The University of Michigan Orthopaedic Research Laboratories Histology Unit 400 North Ingalls, G161 Ann Arbor, MI 48109-0486 my office 734-936-1635 lab office 734-763-9674 Good luck Gayle Callis HTL(ASCP)HT,MT Bozeman MT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jamie E Erickson Sent: Wednesday, May 20, 2009 7:32 AM To: histonet histonet Subject: [Histonet] Sucrose infiltration and Bone question... Hi All, I have two question about sucrose infiltration....First, I have infiltrated many mouse brain with sucrose 30% in the past and it has always been at 4degrees C (dc) and I'm asking myself now why? My mouse brain are fixed in paraformaldehyde or formaldehyde for 24 hours prior so why sucrose @ 4dc? Would it not infiltrate better (faster) at room temp? I could not find out why 4dc... My real problem is that I am tiring to get good mouse tibia/femur frozen sections and I am using D-profile blade and a tape transfer method. It worked great on my rats sections but not mouse so I think the sucrose is not getting in to the bone. I can't use the sinking tissue as an indicator because the bone sinks right away. Should I go longer in sucrose (maybe 48 Hrs) but at 4dc or room temp ? Would adding 2 % DSMO to sucrose facilitate penetration..?? Any suggestions would be great.. Thanks have a great day.. Jamie _______________________________ Jamie Erickson Sr. Research Associate II M.S. HTL (ASCP) Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MadaryJ <@t> MedImmune.com Wed May 20 12:51:15 2009 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Wed May 20 12:51:26 2009 Subject: [Histonet] fite for nocardia In-Reply-To: References: Message-ID: I recall not using just xylene but a mixture of xylene and peanut oil even for the spins the oil is required to coat the bacteria and since there is no paraffin the xylene still acts as kind of a diluent for the peanut oil. Also do not forget ocardia filaments need to be differentiated with sulfuric acid not hydrochloric for best results. At AFIP we called it a Coates Fite after an Air Force guy named Jerry Coates. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, May 20, 2009 1:07 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 66, Issue 23 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Re Nikon Diaphot 300 (Norman, Michael) 2. IHC certification (Achstetter, Virginia A.) 3. Career Opportunity in Chicago area (JimR0712@comcast.net) 4. Looking for career opportunity in Chicago area (JimR0712@comcast.net) 5. Trying to post (af46@buffalo.edu) 6. CLIA regulations (Jonathan De La Rosa) 7. PathNet HLA Module (Brown, Lori) 8. PO/Zylene on cytospins for Fites (Carrie Disbrow) 9. Hi Tony, (Carrie Disbrow) 10. Thanks to those who helped find Larry M. plus (Leroy Brown) 11. Cryoprotection Issues with Unfixed Mouse Hearts (Adam Bazama) 12. RE: Cryoprotection Issues with Unfixed Mouse Hearts (gayle callis) 13. RE: IHC cert. (Lee & Peggy Wenk) 14. Sucrose infiltration and Bone question... (Jamie E Erickson) 15. Histotechnician 2 opportunity in Portland, OR (Fletcher, Annette M) 16. Job Opening In Sacramento, CA (Jennifer McAnally) 17. microwave antigen retrieval (Kimberly Tuttle) 18. (Tissue Processor's) Used, Refurbished or Lease Agreement (Akemi Allison-Tacha) 19. Re: microwave antigen retrieval (rjbuesa@yahoo.com) ---------------------------------------------------------------------- Message: 1 Date: Mon, 18 May 2009 17:00:17 -0400 From: "Norman, Michael" Subject: [Histonet] Re Nikon Diaphot 300 To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <3614638D4C701B438F335F0EFA8F6EBC68C6271112@AACC-MAILBOX.aacc.cc.md.us> Content-Type: text/plain; charset="us-ascii" Does anyone have a .pdf copy of the instructions manual for the Nikon Diaphot 300 Inverted Microscope they could send me? Thanks, Michael Norman ________________________________ The information contained in this email may be confidential and/or legally privileged. It has been sent for the sole use of the intended recipient(s). If the reader of this message is not an intended recipient, you are hereby notified that any unauthorized review, use, disclosure, dissemination, distribution, or copying of this communication, or any of its content, is strictly prohibited. If you have received this communication in error, please contact the sender by reply email and destroy all copies of the original message. Thank you. ------------------------------ Message: 2 Date: Tue, 19 May 2009 07:31:58 -0400 From: "Achstetter, Virginia A." Subject: [Histonet] IHC certification To: Message-ID: <127D612DBCD77546A12C4746F7491C7F053C1AD6@lewis.afip.osd.mil> Content-Type: text/plain; charset="us-ascii" How do I go about finding the study material needed for the IHC exam, how do I prepare, etc. Thanks Ginny Achstetter ------------------------------ Message: 3 Date: Tue, 19 May 2009 17:14:08 +0000 (UTC) From: JimR0712@comcast.net Subject: [Histonet] Career Opportunity in Chicago area To: Histonet2 Message-ID: <387056283.11337581242753248724.JavaMail.root@sz0098a.emeryville.ca.mail.comcast.net> Content-Type: text/plain; charset=utf-8 Last week, my former employer, Loyola University Medical Center in Maywood, Il., layed off 440 employees (about 8 % of the workforce). I was the Manager of Anatomic Pathology (Histo/Cyto/Autopsy/Transcription/MohsLabs/Clerical Support) and now am looking for a career opportunity in the Chicago area. Please let me know if you hear of any career opportunities. Thanks. ------------------------------ Message: 4 Date: Tue, 19 May 2009 17:47:43 +0000 (UTC) From: JimR0712@comcast.net Subject: [Histonet] Looking for career opportunity in Chicago area To: Histonet2 Message-ID: <1482254971.11355411242755263324.JavaMail.root@sz0098a.emeryville.ca.mail.comcast.net> Content-Type: text/plain; charset=utf-8 I am looking for a career opportunity. ----- Forwarded Message ----- From: JimR0712@comcast.net To: "Histonet2" Sent: Tuesday, May 19, 2009 12:14:08 PM GMT -06:00 US/Canada Central Subject: Career Opportunity in Chicago area Last week, my former employer, Loyola University Medical Center in Maywood, Il., layed off 440 employees (about 8 % of the workforce). I was the Manager of Anatomic Pathology (Histo/Cyto/Autopsy/Transcription/MohsLabs/Clerical Support) and now am looking for a career opportunity in the Chicago area. Please let me know if you hear of any career opportunities. Thanks. ------------------------------ Message: 5 Date: Tue, 19 May 2009 14:39:52 -0400 From: af46@buffalo.edu Subject: [Histonet] Trying to post To: Message-ID: <14581.1242758392@buffalo.edu> Content-Type: text/plain; charset="utf-8" just testing to see if I can post ------------------------------ Message: 6 Date: Tue, 19 May 2009 12:45:13 -0700 (PDT) From: Jonathan De La Rosa Subject: [Histonet] CLIA regulations To: Histonet@lists.utsouthwestern.edu Message-ID: <638245.37316.qm@web110701.mail.gq1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello Histoneters, I was wondering if someone could?contact me?offline about CLIA regulations and/or inspections. I have a few concerns. Any help would be appreciated. Thanks in advance. JD jdhisto@yahoo.com ------------------------------ Message: 7 Date: Tue, 19 May 2009 16:08:33 -0400 From: "Brown, Lori" Subject: [Histonet] PathNet HLA Module To: "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: text/plain; charset="us-ascii" I'm hoping to network with some wonderful Pathologists (and informaticists). I'm wondering if anyone out there has ever used or been a part of an implementation of Cerner Millennium's PathNet HLA module for tissue transplant. Any networking and advice is greatly appreciated!! Lori M. Brown, SPHR, CIR Recruiting Manager Vitalize Consulting Solutions | 410-392-5332 o | 484-356-3781 c | 484-734-6168 f | www.getvitalized.com ________________________________ Erase your problems. See how in booth #530 at the International MUSE May 25th - 29th. ________________________________ NOTICE: This message and any included attachments are from VITALIZE CONSULTING SOLUTIONS, INC. and are intended only for the addressee(s). The information contained herein may include intellectual property or privileged or otherwise confidential information. If you are not the named recipient, you are prohibited to forward, print, copy, distribute, disclose, or use such information and your actions may be unlawful. If an addressing or transmission error has misdirected the email, or you have reason to believe you are not authorized to receive it, please promptly delete this message and notify the sender by e-mail with a copy to vcs@getvitalized.com. ------------------------------ Message: 8 Date: Tue, 19 May 2009 17:19:36 -0400 From: "Carrie Disbrow" Subject: [Histonet] PO/Zylene on cytospins for Fites To: Message-ID: <4A12EA2D.72AC.0059.0@shands.ufl.edu> Content-Type: text/plain; charset=US-ASCII Hi Rene, Thanks for the info. We are doing a AFB Fites for Nocardia. There is filamentous bacteria in the patient GMS and the docs are differentiating the type. I did not deparaffinize the patient cytospin slides and I did deparaffinize the patient control with Z/PO. The problem I had been anticipating was that the patient and control did not have the same procedure. I was thinking of it as a CAP issue,,,, but then realized we don't have the same preparation for any stains when using cytospins and controls, ie GMS, Mucin, irons, all have controls that need deparaffinization. DUH! Anyway the stain did not work. It looks like the Methylene blue is too heavy and blocking the carbol fuschin. We use Dako's Artisan stainer, which I love and are having to rework this stain. Thanks again, Carrie ------------------------------ Message: 9 Date: Tue, 19 May 2009 17:22:45 -0400 From: "Carrie Disbrow" Subject: [Histonet] Hi Tony, To: Message-ID: <4A12EAEA.72AC.0059.0@shands.ufl.edu> Content-Type: text/plain; charset=US-ASCII Hi Tony, Thanks for the info. I understand the discussion topic. I just wish their was one straight answer! But then this is histology! Carrie ------------------------------ Message: 10 Date: Tue, 19 May 2009 14:33:22 -0700 From: Leroy Brown Subject: [Histonet] Thanks to those who helped find Larry M. plus To: Message-ID: Content-Type: text/plain; charset="iso-8859-1"; I was able to speak to Larry M. about my tissue processor problems. thank you all for your help. I am now looking for perhaps a broken MVP-1 or 2 since I am no longer able to get the CRT display parts to fix mine. they are interchangeable and if I could find a discarded MVP I could get mine running again. Anyone have a mothballed one sitting in the basement? Thanks LeRoy Brown HT(ASCP)HTL www.histocs.com Everson, Wa 98247 ------------------------------ Message: 11 Date: Tue, 19 May 2009 16:44:08 -0500 From: "Adam Bazama" Subject: [Histonet] Cryoprotection Issues with Unfixed Mouse Hearts To: Message-ID: <000001c9d8ca$f0d14610$d273d230$@edu> Content-Type: text/plain; charset="us-ascii" Does anyone have any suggestions or protocols for cryoprotecting unfixed mouse heart tissue? Some of the ones I have been cryosectioning lately have been damaged, I think because of improper cryoprotection. Thanks! Adam baza0013@umn.edu ------------------------------ Message: 12 Date: Tue, 19 May 2009 16:49:36 -0600 From: "gayle callis" Subject: RE: [Histonet] Cryoprotection Issues with Unfixed Mouse Hearts To: "'Adam Bazama'" , Message-ID: <002101c9d8d4$17177750$454665f0$@callis@bresnan.net> Content-Type: text/plain; charset="us-ascii" Adam, It is not a good to sucrose cryoprotect fresh tissues. Only well fixed tissues should be cryoprotected to lessen freezing artifact caused by large water ice crystals. Charles Scouten, on his website discusses this, and if one cryoprotects fresh tissues, you change the osmolarity within tissues/cells and the tissues shrivel up. This is probably the damage/changes you are observing. Tissues that are well fixed with NBF or a PFA mixture should be sucrose cryoprotected. For fresh tissues, dissect heart from mouse, embed and snap freeze with a proper RAPIC snap freezing method. If you would like, I will privately send a simple snap freezing that eliminates use of precooled isopentane. Avoid freezing in a cryostat or a freezer, even -80C as freezing will be too slow and at warmer temperture than true/rapidsnap freezing methods. This will also cause freezing artifact in both fresh or sucrose cryoprotected fixed tissues. Be sure to read Dr. Scouten's comments on snap freezing and its effects on tissues - it is very informative. Gayle Callis HTL(ASCP)HT,MT Bozeman MT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Adam Bazama Sent: Tuesday, May 19, 2009 3:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cryoprotection Issues with Unfixed Mouse Hearts Does anyone have any suggestions or protocols for cryoprotecting unfixed mouse heart tissue? Some of the ones I have been cryosectioning lately have been damaged, I think because of improper cryoprotection. Thanks! Adam baza0013@umn.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Tue, 19 May 2009 20:19:46 -0400 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] IHC cert. To: "'Ginny .'" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" For all information about certification exams (e.g., HT, HTL, SLS (specialist in laboratory safety), or on qualification exams (e.g., QIHC), go to www.ascp.org Click on Laboratory Professional Click on Certification Click on either "Get Certified" or "Get Qualified" Under Get Qualified, there is information on study material (#5) - Reading list - Topic outline Peggy A. Wenk, HTL(ASCP)SLS Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ginny . Sent: Tuesday, May 19, 2009 7:37 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC cert. Where can I find the study material needed to prepare for the IHC exam?? When on the ASCP website but found nothing.? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Wed, 20 May 2009 09:32:09 -0400 From: Jamie E Erickson Subject: [Histonet] Sucrose infiltration and Bone question... To: "histonet histonet" Message-ID: Content-Type: text/plain; charset="US-ASCII" Hi All, I have two question about sucrose infiltration....First, I have infiltrated many mouse brain with sucrose 30% in the past and it has always been at 4degrees C (dc) and I'm asking myself now why? My mouse brain are fixed in paraformaldehyde or formaldehyde for 24 hours prior so why sucrose @ 4dc? Would it not infiltrate better (faster) at room temp? I could not find out why 4dc... My real problem is that I am tiring to get good mouse tibia/femur frozen sections and I am using D-profile blade and a tape transfer method. It worked great on my rats sections but not mouse so I think the sucrose is not getting in to the bone. I can't use the sinking tissue as an indicator because the bone sinks right away. Should I go longer in sucrose (maybe 48 Hrs) but at 4dc or room temp ? Would adding 2 % DSMO to sucrose facilitate penetration..?? Any suggestions would be great.. Thanks have a great day.. Jamie _______________________________ Jamie Erickson Sr. Research Associate II M.S. HTL (ASCP) Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com ------------------------------ Message: 15 Date: Tue, 19 May 2009 11:23:22 -0700 From: "Fletcher, Annette M" Subject: [Histonet] Histotechnician 2 opportunity in Portland, OR To: Message-ID: <284596E350CB0743BB500B18475E7A0B022A6562@wn1223.or.providence.org> Content-Type: text/plain; charset="iso-8859-1" Good Afternoon Histonetters, Annette Fletcher here with Providence Health and Services in Portland, Oregon. We are seeking a Histotech for our Regional Lab in Portland, OR. This is an opportunity to work for a visionary Manager who is dedicated to growing and developing her Histo staff. We have a state of the art lab, highly competitive compensation and an excellent benefit plan. We also offer relocation assistance. You can email me at annette.fletcher@providence.org with your interest. Thank you, Annette M Fletcher Senior Recruiter Providence Health & Services, Employment 1235 NE 47th Avenue - Suite 200 Portland, OR 97213 T: 503-215-5840 F: 503-215-5770 Annette.fletcher@providence.org Confidentiality notice: The information contained in this email message including attachments is confidential and is intended only for the use of the individual or entity named above and others who have been specifically authorized to receive it. If you are not the intended recipient, you are hereby notified that any use, unauthorized dissemination, distribution, or copying of this communication is strictly prohibited. If you have received this communication in error, please delete immediately or if any problems occur with transmission, please notify me immediately by telephone at 503-215-5840 or by reply e-mail at annette.fletcher@providence.org. Thank you! DISCLAIMER: This message is intended for the sole use of the addressee, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If you are not the addressee you are hereby notified that you may not use, copy, disclose, or distribute to anyone the message or any information contained in the message. If you have received this message in error, please immediately advise the sender by reply email and delete this message. ------------------------------ Message: 16 Date: Wed, 20 May 2009 09:32:11 -0500 From: "Jennifer McAnally" Subject: [Histonet] Job Opening In Sacramento, CA To: , "'LMargraf@childmed.dallas.tx.us.'" Message-ID: Content-Type: text/plain; charset="us-ascii" ** *Hello Histonetters,* ** *I have an opening for a Lead/Supervisory Histotechnologist in Sacramento, CA. * ** * Please review the job description below. If you know someone qualified for this position, please forward this information. For anyone interested, please forward your resume to jennifer@phcconsulting.com for consideration. This Lab is part of a multi-lab company. This particular Lab processes around 300 cases a day. This person will be accountable for the direct supervision and operation of the lab and staff. We need someone who can support the company's growth oriented business plan and has an entrepreneurial spirit. This person will be vital to the overall success of the lab so the following characteristics are necessary: Great leadership skills, Ability to train others, Able to smoothly manage the workflow of the lab, Great attitude, energetic and pleasant personality. 3-5 years experience as a Histotechnologist, with at least 1-2 years in leadership role preferred. Salary range is $70k to $95k depending on experience. Performance bonuses, full benefits, healthcare, health savings accounts, 401k and accrued vacation. Relo package offered for the right person. Jennifer McAnally PHC Consulting 888-263-5688 ext 300 ------------------------------ Message: 17 Date: Wed, 20 May 2009 11:26:32 -0400 From: "Kimberly Tuttle" Subject: [Histonet] microwave antigen retrieval To: "histonet" Message-ID: <4A13E8E7.90CE.001A.3@umm.edu> Content-Type: text/plain; charset=US-ASCII Can someone post instructions for microwave antigen retrieval... my pressure cooker broke and I need an immediate solution. Kimberly C. Tuttle HT (ASCP) Pathology Biorepository and Research Core University of Maryland Room NBW58, UMMC 22 S. Greene St Baltimore, MD 21201 (410) 328-5524 (410) 328-5508 fax This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. ------------------------------ Message: 18 Date: Wed, 20 May 2009 08:34:20 -0700 (PDT) From: Akemi Allison-Tacha Subject: [Histonet] (Tissue Processor's) Used, Refurbished or Lease Agreement To: histonet Message-ID: <780411.19431.qm@web31301.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Good Morning out there in Histo Land! Calling all Vendors. ?I have recently accepted a new position as the Histology Manager at APMG Labs in Los Gatos, CA. ?We are down one Tissue Processor. ?The Executive Administrator thought we would be purchasing a used VIP from one of our affiliates, but unfortunately this possibility has fallen through. ? I would appreciate any Vendors who have Tissue Processor's for sale that are Used or Refurbished. ?I am also open to the possibility of?negotiating?lease agreements. ?I realize warranties and service agreements are extremely important. Please e-mail me any available options. ? Thanks,Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Ct. Los Gatos, CA 95032 W: (408)884.2718? W: E-Mail: aallison-tacha@apmglab.com P: E-Mail: akemiat3377@yahoo.com ------------------------------ Message: 19 Date: Wed, 20 May 2009 09:05:18 -0700 (PDT) From: rjbuesa@yahoo.com Subject: Re: [Histonet] microwave antigen retrieval To: histonet , Kimberly Tuttle Message-ID: <848945.6183.qm@web65710.mail.ac4.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 You use the MW to heat the HIER solution and the slides in it. Once the buffer boiled with the slides, remove the container with the slides and place it in either a water bath or a steamer at 98?C during 20 minutes. After that place the container over the counter during another 20 minutes. Wash with PBS and start the staining procedure (either manual or automated). Ren? J. --- On Wed, 5/20/09, Kimberly Tuttle wrote: From: Kimberly Tuttle Subject: [Histonet] microwave antigen retrieval To: "histonet" Date: Wednesday, May 20, 2009, 11:26 AM Can someone post instructions for microwave antigen retrieval... my pressure cooker broke and I need an immediate solution. Kimberly C. Tuttle? HT (ASCP) Pathology Biorepository and Research Core University of Maryland Room NBW58, UMMC 22 S. Greene St Baltimore, MD 21201 (410) 328-5524 (410) 328-5508 fax This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 66, Issue 23 **************************************** To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary. This communication is expected to be read and/or used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation. From jtaylor <@t> meriter.com Wed May 20 14:06:26 2009 From: jtaylor <@t> meriter.com (Taylor, Jean) Date: Wed May 20 14:06:31 2009 Subject: [Histonet] CMV antibody Message-ID: <466B666475DE6547BBB0641E540A4BB5048889AE2E@EXVS1.meriter.com> Hi Everyone, I'm wondering what antibody clone labs are using to detect Cytomegalovirus by IHC. Thank you! Jean Taylor, HT(ASCP) QIHC IHC Tech. Meriter Health Services Madison, WI 53715 From azdudley <@t> hotmail.com Wed May 20 14:32:41 2009 From: azdudley <@t> hotmail.com (anita dudley) Date: Wed May 20 14:32:45 2009 Subject: [Histonet] richard allan dehydrant Message-ID: just wondering if anyone had richard allans 95 and 100 dehydrant on the processor and if they had any problems. I have switched from flex, our cytology dept uses the dehydrant and we share the same room. anyway our little bxs looked a little fried today, the oven was up a little also so it might have been that. would appreciate any input on the dehydrant. thanks anita dudley providence hosp mobile, al 36695 _________________________________________________________________ Insert movie times and more without leaving Hotmail?. http://windowslive.com/Tutorial/Hotmail/QuickAdd?ocid=TXT_TAGLM_WL_HM_Tutorial_QuickAdd1_052009 From burch007 <@t> mc.duke.edu Wed May 20 14:54:15 2009 From: burch007 <@t> mc.duke.edu (James L Burchette) Date: Wed May 20 14:54:24 2009 Subject: [Histonet] Re: [IHCRG] CMV antibody In-Reply-To: <466B666475DE6547BBB0641E540A4BB5048889AE2E@EXVS1.meriter.com> Message-ID: Dear Jean, I am very fond of Dako's M0854 and find I can use it at 1:400-1:500 when pepsin is used as a pretreatment proteolytic enzyme as compared to HIER pretreatment with a citrate buffer. CMV is run everyday in my lab. Jim Burchette "Taylor, Jean" Sent by: ihcrg@googlegroups.com 05/20/2009 03:06 PM Please respond to JTAYLOR@meriter.com To "'ihcrg@googlegroups.com'" , "'histonet@lists.utsouthwestern.edu'" cc Subject [IHCRG] CMV antibody Hi Everyone, I?m wondering what antibody clone labs are using to detect Cytomegalovirus by IHC. Thank you! Jean Taylor, HT(ASCP) QIHC IHC Tech. Meriter Health Services Madison, WI 53715 From jtaylor <@t> meriter.com Wed May 20 15:33:55 2009 From: jtaylor <@t> meriter.com (Taylor, Jean) Date: Wed May 20 15:33:59 2009 Subject: [Histonet] CMV Message-ID: <466B666475DE6547BBB0641E540A4BB5048889AE2F@EXVS1.meriter.com> Hi Everyone, The pathologist that I work with wanted me to add that the CMV that we're staining for is in the GI tract, and he's wondering if there is a different clone/procedure that labs use for this? Thanks again!! Jean Taylor, HT(ASCP) QIHC IHC Tech Meriter Health Services Madison, WI 53715 From Reuel.Cornelia <@t> tsrh.org Wed May 20 15:52:40 2009 From: Reuel.Cornelia <@t> tsrh.org (Reuel Cornelia) Date: Wed May 20 15:53:10 2009 Subject: [Histonet] Gross photography/macrophotography Message-ID: <4A142748020000C50005794C@nwcl02.tsrh.org> Hello histonetters, I am looking for a good digital camera for gross photography. Any recommendations that works with your lab will be beneficial. Thank you. Happy memorial day!!! Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 From joseph-galbraith <@t> uiowa.edu Wed May 20 16:12:58 2009 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Wed May 20 16:13:15 2009 Subject: [Histonet] Leica cryostat In-Reply-To: References: Message-ID: Janet: I've worked with the CM1850 and 1950 and like them a lot. You listed the 1800 twice. Was one of your options an 1850? Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dertien, Janet Sent: Wednesday, May 13, 2009 3:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Leica cryostat Hello everyone, We have decided to purchase a refurbished Leica cryostat and are trying to decide which model to choose; does anyone have experience with the 1510, the CM 1800 or the 1800? Any feedback would be appreciated. Thanks! Janet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Wed May 20 16:15:00 2009 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Wed May 20 16:13:44 2009 Subject: [Histonet] Gross photography/macrophotography In-Reply-To: <4A142748020000C50005794C@nwcl02.tsrh.org> References: <4A142748020000C50005794C@nwcl02.tsrh.org> Message-ID: <4A1472D4.5090006@umdnj.edu> Most digital cameras from the well-known manufacturers (Nikon, Canon, Pentax, Olympus) will be fine for gross photography. An single lens reflex (SLR) type of camera with interchangable lenses is ideal but I suggest NOT buying the zoom lens in a "kit" but buy camera body and a macro lens that will allow close focusing. The macro lens should have a focal length of 90 mm or longer to allow for comfortable working distances. One example would be a Nikon D5000 and a 105 mm Micro Nikor lens. Instead of the Nikor lens a Tamron 90 mm lens is half as expensive and will give fine results. A digital SLR + macro lens will be over $1000. Even a small "point and shoot" camera will do the job. I photograph gross anatomy exams with a little Canon I can fit in my pocket. Do not be misled by the "megapixel wars", more megapixels does not make a better camera. The lens and the processor are just as important. I'm sure there is a professional level camera store (not Best Buy) in Dallas that can help you make a decision. Geoff Reuel Cornelia wrote: > Hello histonetters, > I am looking for a good digital camera for gross photography. Any recommendations that works with your lab will be beneficial. Thank you. Happy memorial day!!! > > > > Reuel Cornelia, BS MT, AMT > Cellular Pathology > Texas Scottish Rite Hospital for Children > 2222 Welborn Street > Dallas, TX 75219 > Tel: 214-559-7766 > fax: 214-559-7768 > > > > _______________________________________________ > 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 mcauliff@umdnj.edu ********************************************** From joseph-galbraith <@t> uiowa.edu Wed May 20 17:32:50 2009 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Wed May 20 17:32:54 2009 Subject: [Histonet] Semiautomated Antigen Retrival systems/equipment Message-ID: Hello all: Is anyone using any standalone semi-automated antigen retrieval equipment? I've seen a few microwave or wet bath based systems (sometimes integrated into full blown IHC staining systems) in the lit and on the net. How many people are still using the Nordicware pressure cooker in a microwave or the BioCare Medical electric antigen retrieval pressure cooker? Comments are welcome as we are looking at short and long term solutions to make the IHC process less labor intensive. Thanks. Joe From Annette.Fletcher <@t> providence.org Wed May 20 17:39:34 2009 From: Annette.Fletcher <@t> providence.org (Fletcher, Annette M) Date: Wed May 20 17:40:31 2009 Subject: [Histonet] Histonet Opportunity Message-ID: <284596E350CB0743BB500B18475E7A0B022A6592@wn1223.or.providence.org> Hi Histonetters, Providence Health and Services in Portland, OR is seeking a Histotech to join their team in a high-volume, state-of-the-art lab. The Histotech will be supported by an empowering manager who encourages employee development and growth. Employee satisfaction scores in this department are among the highest in the Hospital. Our compensation is highly competitive and we have excellent benefits. We do offer relocation assistance. No agency's please. If you are interested in learning more about this opportunity, please contact me at 503-215-5840 or annette.fletcher@providence.org Annette M Fletcher Senior Recruiter Providence Health & Services, Employment 1235 NE 47th Avenue - Suite 200 Portland, OR 97213 T: 503-215-5840 F: 503-215-5770 Annette.fletcher@providence.org DISCLAIMER: This message is intended for the sole use of the addressee, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If you are not the addressee you are hereby notified that you may not use, copy, disclose, or distribute to anyone the message or any information contained in the message. If you have received this message in error, please immediately advise the sender by reply email and delete this message. From AnthonyH <@t> chw.edu.au Wed May 20 18:12:26 2009 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Wed May 20 18:12:36 2009 Subject: [Histonet] RE: [IHCRG] CMV In-Reply-To: <466B666475DE6547BBB0641E540A4BB5048889AE2F@EXVS1.meriter.com> Message-ID: Jean, No the virus is the same for IPX purposes. I have used both the Dako (Clones CCH2 + DDG9) and the Novocastra products (Clones 2 and 6) and they both stain the same Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: ihcrg@googlegroups.com [mailto:ihcrg@googlegroups.com] On Behalf Of Taylor, Jean Sent: Thursday, 21 May 2009 6:34 AM To: 'ihcrg@googlegroups.com'; 'histonet@lists.utsouthwestern.edu' Subject: [IHCRG] CMV Hi Everyone, The pathologist that I work with wanted me to add that the CMV that we're staining for is in the GI tract, and he's wondering if there is a different clone/procedure that labs use for this? Thanks again!! Jean Taylor, HT(ASCP) QIHC IHC Tech Meriter Health Services Madison, WI 53715 ********************************************************************* 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 note 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 kmerriam2003 <@t> yahoo.com Thu May 21 07:41:44 2009 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Thu May 21 07:41:49 2009 Subject: [Histonet] HRP-labeled primary antibodies Message-ID: <905854.25089.qm@web50310.mail.re2.yahoo.com> Hi?All, Has anyone had experience doing IHC on FFPE tissues with HRP-labeled primary antibodies?? I?was wondering what the best way to detect them would be.? I assume that going strait to DAB would not work, since no amplification is there.??I was thinking of using a biotinyl tyramide step to amplify the signal. Also, do you think the final antibody concentration would?need to be higher than with traditional, unlabeled primaries? Thanks, Kim ?Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA From je2 <@t> sanger.ac.uk Thu May 21 07:42:14 2009 From: je2 <@t> sanger.ac.uk (Jeanne Estabel) Date: Thu May 21 07:42:21 2009 Subject: [Histonet] slide scanner Message-ID: <8DAC27FBBF33764B9F3DCF0A3E6B096202CAC134@exchsrv2.internal.sanger.ac.uk> Hi, I am looking to buy a slide scanner for highthrouput project. Could you give me your opinion if you are using one of the next 3: Mirax Scan Zeiss, Nanozoomer Hamamatsu and XT Aperio? Best regards Jeanne Jeanne Estabel, PhD MGP Histology Operations Manager Wellcome Trust/Sanger Institute UK -- The Wellcome Trust Sanger Institute is operated by Genome Research Limited, a charity registered in England with number 1021457 and a compa ny registered in England with number 2742969, whose registered office is 2 15 Euston Road, London, NW1 2BE. From asmith <@t> mail.barry.edu Thu May 21 08:03:23 2009 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Thu May 21 08:05:35 2009 Subject: [Histonet] Gross photography/macrophotography In-Reply-To: <4A142748020000C50005794C@nwcl02.tsrh.org> References: <4A142748020000C50005794C@nwcl02.tsrh.org> Message-ID: I have been very happy with the Olympus C-7000. It will focus at 3 1/4 inches (8 cm) in its macro mode. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Reuel Cornelia Sent: Wednesday, May 20, 2009 4:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Gross photography/macrophotography Hello histonetters, I am looking for a good digital camera for gross photography. Any recommendations that works with your lab will be beneficial. Thank you. Happy memorial day!!! Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From redward <@t> ucb.br Thu May 21 08:46:59 2009 From: redward <@t> ucb.br (Robert Edward Pogue) Date: Thu May 21 08:41:34 2009 Subject: [Histonet] Bone! Message-ID: <0C9E32CEFAAF894CBE806BEE2206F75E02A16AAA@supra.ucb.br> Friends, Can someone recommend to me how to store bones (juvenile rat femur) that I have fixed and decalcified, but am not yet ready to cut (they're for vibratome, so not embedded). Thanks! Redward. From NMargaryan <@t> childrensmemorial.org Thu May 21 09:35:11 2009 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Thu May 21 09:37:05 2009 Subject: [Histonet] bone fixation and calcification Message-ID: Dear Histonetters, I am new in fixation and processing bones. I need your full protocol with details how to fix and process mice bone to visualize the tumor metastases? Thanks in advance, Naira From anh2006 <@t> med.cornell.edu Thu May 21 09:48:15 2009 From: anh2006 <@t> med.cornell.edu (anh2006@med.cornell.edu) Date: Thu May 21 09:49:20 2009 Subject: [Histonet] HRP-labeled primary antibodies In-Reply-To: <905854.25089.qm@web50310.mail.re2.yahoo.com> References: <905854.25089.qm@web50310.mail.re2.yahoo.com> Message-ID: <378114708-1242917353-cardhu_decombobulator_blackberry.rim.net-1448245432-@bxe1087.bisx.prod.on.blackberry> If there is enough of the antigen in the tissue or sample you can detect without amplification. HRP/DAB is an enzymatic reaction so it is also is amplification. I would try it the straight up way before diving into more complex protocols. Alternatively another option would be that you could try to come in with a secondary antibody (either HRP labeled itself or biotinylated). If your secondary is a polyclonal - which most are - it should still be able to detect the primary even with the HRP attached. Worth trying anyway. -----Original Message----- From: Kim Merriam Date: Thu, 21 May 2009 05:41:44 To: Histonet; Subject: [Histonet] HRP-labeled primary antibodies Hi?All, Has anyone had experience doing IHC on FFPE tissues with HRP-labeled primary antibodies?? I?was wondering what the best way to detect them would be.? I assume that going strait to DAB would not work, since no amplification is there.??I was thinking of using a biotinyl tyramide step to amplify the signal. Also, do you think the final antibody concentration would?need to be higher than with traditional, unlabeled primaries? Thanks, Kim ?Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kmerriam2003 <@t> yahoo.com Thu May 21 09:52:14 2009 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Thu May 21 09:52:20 2009 Subject: [Histonet] HRP-labeled primary antibodies In-Reply-To: <378114708-1242917353-cardhu_decombobulator_blackberry.rim.net-1448245432-@bxe1087.bisx.prod.on.blackberry> References: <905854.25089.qm@web50310.mail.re2.yahoo.com> <378114708-1242917353-cardhu_decombobulator_blackberry.rim.net-1448245432-@bxe1087.bisx.prod.on.blackberry> Message-ID: <237236.38369.qm@web50308.mail.re2.yahoo.com> I got some great ideas.? Thanks everyone! Kim ?Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ________________________________ From: "anh2006@med.cornell.edu" To: Kim Merriam ; Histonet ; ihcrg@googlegroups.com Sent: Thursday, May 21, 2009 10:48:15 AM Subject: Re: [Histonet] HRP-labeled primary antibodies If there is enough of the antigen in the tissue or sample you can detect without amplification. HRP/DAB is an enzymatic reaction so it is also is amplification. I would try it the straight up way before diving into more complex protocols. Alternatively another option would be that you could try to come in with a secondary antibody (either HRP labeled itself or biotinylated). If your secondary is a polyclonal - which most are - it should still be able to detect the primary even with the HRP attached. Worth trying anyway. -----Original Message----- From: Kim Merriam Date: Thu, 21 May 2009 05:41:44 To: Histonet; Subject: [Histonet] HRP-labeled primary antibodies Hi?All, Has anyone had experience doing IHC on FFPE tissues with HRP-labeled primary antibodies?? I?was wondering what the best way to detect them would be.? I assume that going strait to DAB would not work, since no amplification is there.??I was thinking of using a biotinyl tyramide step to amplify the signal. Also, do you think the final antibody concentration would?need to be higher than with traditional, unlabeled primaries? Thanks, Kim ?Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Reuel.Cornelia <@t> tsrh.org Thu May 21 10:12:23 2009 From: Reuel.Cornelia <@t> tsrh.org (Reuel Cornelia) Date: Thu May 21 10:12:57 2009 Subject: [Histonet] bone tissue Message-ID: <4A152907020000C500057A76@nwcl02.tsrh.org> Can somebody help me with my bone tissue to remain intact during antigen retrieval for IHC staining. I have used subbed slides but It did not help me. Is there more better adhesive. Does Haupts Gelatin works better? I know this is a hundred year old question and hopefully this problem have been resolved since my undated knowledge could remember. I would like to thank everybody for responding to my last e-mail on gross photography. It gives me more idea what to purchase. Thanks again histonetters and I really appreciate your help. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 From redward <@t> ucb.br Thu May 21 10:33:08 2009 From: redward <@t> ucb.br (Robert Edward Pogue) Date: Thu May 21 10:28:33 2009 Subject: [Histonet] Bone! References: <86ADE4EB583CE64799A9924684A0FBBF06C0D289@wahtntex2.waht.swest.nhs.uk> Message-ID: <0C9E32CEFAAF894CBE806BEE2206F75E02A16AAB@supra.ucb.br> Thanks Kemlo... I thought of that, but was worried about over-fixation,- what do you think? Redward. ------------------------------ In 10% neutral buffered formalin? Kemlo Rogerson e-mail kemlorogerson@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert Edward Pogue Sent: 21 May 2009 14:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone! Friends, Can someone recommend to me how to store bones (juvenile rat femur) that I have fixed and decalcified, but am not yet ready to cut (they're for vibratome, so not embedded). Thanks! Redward. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CBark <@t> memorialcare.org Thu May 21 11:05:50 2009 From: CBark <@t> memorialcare.org (Christine Bark) Date: Thu May 21 11:07:22 2009 Subject: [Histonet] Bone saw Message-ID: Good Morning, I was wondering if anyone can help me find a really good saw for bones (femoral/humeral heads mainly). We currently have a MarMed bone saw that works great for knees and such but it's just not strong enough for the femurs. Thank you, Christine Bark HT(ASCP) CM Lead Histotech, Pathology Saddleback Memorial Medical Center 949-452-3548 cbark@memorialcare.org ______________________________________________________________________________ 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 anh2006 <@t> med.cornell.edu Thu May 21 11:16:17 2009 From: anh2006 <@t> med.cornell.edu (Andrea Hooper) Date: Thu May 21 11:16:24 2009 Subject: [Histonet] MCP-1/3 Message-ID: Anyone have any good antibodies to recommend for MCP-1 and MCP-3? THANKS! -- From leiker <@t> buffalo.edu Thu May 21 11:20:17 2009 From: leiker <@t> buffalo.edu (Merced M Leiker) Date: Thu May 21 11:20:26 2009 Subject: [Histonet] immunophenotyping pigs In-Reply-To: <905854.25089.qm@web50310.mail.re2.yahoo.com> References: <905854.25089.qm@web50310.mail.re2.yahoo.com> Message-ID: <3D8B70934FF500CE917904B9@CDYwxp1931.ad.med.buffalo.edu> Anyone know how to immunophenotype pigs? Or any published references? Thanks... Merced M Leiker Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 leiker@buffalo.edu 716-829-6118 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. From CBraaten <@t> Cheshire-Med.COM Thu May 21 11:41:35 2009 From: CBraaten <@t> Cheshire-Med.COM (Christine I. Braaten) Date: Thu May 21 11:41:41 2009 Subject: [Histonet] cloudy formalin in tissue processor Message-ID: Has anyone had a problem with their formalin getting cloudy in their tissue processor? We have a VIP tissue processor and just recently started having problems with cloudy formalin. Any suggestions would be appreciated. I checked the archives and couldn't find any articles on this particular subject. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients 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 electronic mail and destroy all copies of the original message. From ratliffjack <@t> hotmail.com Thu May 21 11:41:37 2009 From: ratliffjack <@t> hotmail.com (Jack Ratliff) Date: Thu May 21 11:41:43 2009 Subject: [Histonet] Bone saw In-Reply-To: References: Message-ID: IMEB also has a Bone Band Saw. http://www.imebinc.com/Item/BBS-82203.htm Jack > Date: Thu, 21 May 2009 09:05:50 -0700 > From: CBark@memorialcare.org > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Bone saw > > Good Morning, > I was wondering if anyone can help me find a really good saw > for bones (femoral/humeral heads mainly). We currently have a MarMed > bone saw that works great for knees and such but it's just not strong > enough for the femurs. > Thank you, > > Christine Bark HT(ASCP) CM > Lead Histotech, Pathology > Saddleback Memorial Medical Center > 949-452-3548 > cbark@memorialcare.org > > > > ______________________________________________________________________________ > 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. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ratliffjack <@t> hotmail.com Thu May 21 11:45:39 2009 From: ratliffjack <@t> hotmail.com (Jack Ratliff) Date: Thu May 21 11:45:43 2009 Subject: [Histonet] bone tissue In-Reply-To: <4A152907020000C500057A76@nwcl02.tsrh.org> References: <4A152907020000C500057A76@nwcl02.tsrh.org> Message-ID: Haupts Gelatin works really well at a 1:1 ratio (Concentrate:50% EtOH). The only limitation is background staining from say a hematoxylin counterstain after the IHC. You can purchase the concentrate from Fisher (Haupts Fixative #785-71). Jack > Date: Thu, 21 May 2009 10:12:23 -0500 > From: Reuel.Cornelia@tsrh.org > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] bone tissue > > Can somebody help me with my bone tissue to remain intact during antigen retrieval for IHC staining. I have used subbed slides but It did not help me. Is there more better adhesive. Does Haupts Gelatin works better? I know this is a hundred year old question and hopefully this problem have been resolved since my undated knowledge could remember. > I would like to thank everybody for responding to my last e-mail on gross photography. It gives me more idea what to purchase. Thanks again histonetters and I really appreciate your help. > > > > Reuel Cornelia, BS MT, AMT > Cellular Pathology > Texas Scottish Rite Hospital for Children > 2222 Welborn Street > Dallas, TX 75219 > Tel: 214-559-7766 > fax: 214-559-7768 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ratliffjack <@t> hotmail.com Thu May 21 11:54:20 2009 From: ratliffjack <@t> hotmail.com (Jack Ratliff) Date: Thu May 21 11:54:25 2009 Subject: [Histonet] Bone! In-Reply-To: <0C9E32CEFAAF894CBE806BEE2206F75E02A16AAA@supra.ucb.br> References: <0C9E32CEFAAF894CBE806BEE2206F75E02A16AAA@supra.ucb.br> Message-ID: I do not routinely perform decalcified bone preparations as I predominantly utilize undemineralized resin (MMA) applications, so maybe someone else can better respond to this question. However, I would say that you could store your bone in 70% EtOH until you are ready to process, but the length of time in solution storage may cause you problems down the road since you have already decalcified. I think that a prolonged storage time might make your bone hard again....if that makes any sense....and cause you a fit later when you go to the microtome. Typically, I store undemineralized bones at either 10% NBF or 70% EtOH depending upon what I care to see at the microscope. Jack > Date: Thu, 21 May 2009 10:46:59 -0300 > From: redward@ucb.br > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Bone! > > Friends, > > Can someone recommend to me how to store bones (juvenile rat femur) that I have fixed and decalcified, but am not yet ready to cut (they're for vibratome, so not embedded). > > Thanks! > > Redward. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu May 21 11:55:45 2009 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu May 21 11:55:46 2009 Subject: [IHCRG] Re: [Histonet] HRP-labeled primary antibodies In-Reply-To: <237236.38369.qm@web50308.mail.re2.yahoo.com> References: <905854.25089.qm@web50310.mail.re2.yahoo.com> <378114708-1242917353-cardhu_decombobulator_blackberry.rim.net-1448245432-@bxe1087.bisx.prod.on.blackberry> <237236.38369.qm@web50308.mail.re2.yahoo.com> Message-ID: I agree with Kim, if I was worried about a weak signal using abs directly conjugated to hrp I would just ignore the fact that they have the hrp and use a detection such as labeled polymer matched to the species of the primary ab just like I would without the direct hrp conjugation, there should still be enough sites left on the primary antibody for a secondary detection reagent to attach to the species ab even with some being taken up by the hrp. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _____ From: ihcrg@googlegroups.com [mailto:ihcrg@googlegroups.com] On Behalf Of Kim Merriam Sent: Thursday, May 21, 2009 8:52 AM To: Histonet; ihcrg@googlegroups.com Subject: [IHCRG] Re: [Histonet] HRP-labeled primary antibodies I got some great ideas. Thanks everyone! Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA _____ From: "anh2006@med.cornell.edu" To: Kim Merriam ; Histonet ; ihcrg@googlegroups.com Sent: Thursday, May 21, 2009 10:48:15 AM Subject: Re: [Histonet] HRP-labeled primary antibodies If there is enough of the antigen in the tissue or sample you can detect without amplification. HRP/DAB is an enzymatic reaction so it is also is amplification. I would try it the straight up way before diving into more complex protocols. Alternatively another option would be that you could try to come in with a secondary antibody (either HRP labeled itself or biotinylated). If your secondary is a polyclonal - which most are - it should still be able to detect the primary even with the HRP attached. Worth trying anyway. -----Original Message----- From: Kim Merriam Date: Thu, 21 May 2009 05:41:44 To: Histonet; Subject: [Histonet] HRP-labeled primary antibodies Hi All, Has anyone had experience doing IHC on FFPE tissues with HRP-labeled primary antibodies? I was wondering what the best way to detect them would be. I assume that going strait to DAB would not work, since no amplification is there. I was thinking of using a biotinyl tyramide step to amplify the signal. Also, do you think the final antibody concentration would need to be higher than with traditional, unlabeled primaries? Thanks, Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Montina.VanMeter <@t> pbrc.edu Thu May 21 12:10:59 2009 From: Montina.VanMeter <@t> pbrc.edu (Montina Van Meter) Date: Thu May 21 12:18:15 2009 Subject: [Histonet] fume hood regulations Message-ID: <4FE7FB862E90E448AE32388E759220E50140AE1C@pbrcas31.pbrc.edu> Fellow Histonetters, Does anyone have knowledge of guidelines or regulations on whether a fume hood must remain "on" 24/7? In a cost cutting measure a facility is considering turning off the fume hoods from 7pm to 5am. I have expressed my concern with the idea because of the huge health and safety issue for those who work after hours. This facility could be setting themselves up for a huge liability law suit. I would appreciate written guidelines to strengthen my opposition to this "cost cutting " measure. Thank you, Tina From Lynn.Burton <@t> Illinois.gov Thu May 21 12:23:29 2009 From: Lynn.Burton <@t> Illinois.gov (Burton, Lynn) Date: Thu May 21 12:24:24 2009 Subject: [Histonet] cloudy formalin in tissue processor References: Message-ID: Is your formalin picking up water from the air in the room? Check the humidity level in your room. Lynn Burton Lab Assoc. I Animal Disease Lab Galesburg, Il 61401 ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Christine I. Braaten Sent: Thu 5/21/2009 11:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cloudy formalin in tissue processor Has anyone had a problem with their formalin getting cloudy in their tissue processor? We have a VIP tissue processor and just recently started having problems with cloudy formalin. Any suggestions would be appreciated. I checked the archives and couldn't find any articles on this particular subject. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients 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 electronic mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From petepath <@t> yahoo.com Thu May 21 12:36:10 2009 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Thu May 21 12:36:14 2009 Subject: [Histonet] RE :Bone saw Message-ID: <261651.68516.qm@web45103.mail.sp1.yahoo.com> Hi Christine, About 5 years ago I?modified the jaws of an ?angle vise?for holding bones and attached it to a platform? for use in our laboratory. I found an ideal over the counter saw called a Bosche power hand saw which cost about $100. When?I brought it to our lead PA ?he returned to me and said it took 20 minutes to do the job that took two hours with his previous bone cutting saw. After about 4 years of great success with this pair I decided to?make a small run of??vise designed to be used to hold bones for sawing. I recommend this same inexpensive saw or any hand saw to use with it. You can serial section a femoral head in many 2-3 mm slices effortless. If you visit my web site at: ?http://www.pathologyinnovations.com/bone_vise.htm you can see the saw and vice in video and pictures. Please contact me if you have any further questions. Stephen Peters M.D.? Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 Phone and fax 201 847 7600 www.pathologyinnovations.com From ynwang <@t> u.washington.edu Thu May 21 13:16:54 2009 From: ynwang <@t> u.washington.edu (Yak-Nam Wang) Date: Thu May 21 13:17:18 2009 Subject: [Histonet] fume hood regulations In-Reply-To: <4FE7FB862E90E448AE32388E759220E50140AE1C@pbrcas31.pbrc.edu> Message-ID: Hi, At the University of Washington we have been told to lower the sash all the way when the hood is not in use to save energy and money. I believe there have been several Universities that have done some cost analysis of this and have show large savings in money (one study stated $1500 per year per fume hood in savings and a reduction in 10,600 lbs of CO2 emissions). If you want more information just do a search for "shut the sash". So, if it is not feasible to shut off the hood you can save energy and money just by closing the sash. Have a great weekend. Yak-Nam On 5/21/09 10:10 AM, "Montina Van Meter" wrote: > Fellow Histonetters, > Does anyone have knowledge of guidelines or regulations on whether a > fume hood must remain "on" 24/7? In a cost cutting measure a facility > is considering turning off the fume hoods from 7pm to 5am. I have > expressed my concern with the idea because of the huge health and safety > issue for those who work after hours. This facility could be setting > themselves up for a huge liability law suit. I would appreciate > written guidelines to strengthen my opposition to this "cost cutting " > measure. > > Thank you, > > Tina > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tdobersztyn <@t> chmca.org Thu May 21 13:19:22 2009 From: tdobersztyn <@t> chmca.org (tdobersztyn@chmca.org) Date: Thu May 21 13:19:36 2009 Subject: [Histonet] IHC question- freezing aliquots In-Reply-To: <20090521170214.404C21D790A@interceptor.chmca.org> Message-ID: Hello all! I have an antibody that has a "2" month expiration date. It is recommended that we aliquot, and freeze (-80) the antibody- per the vendor. How do you interpret: The CAP reg pertaining to expired antibody use: ANP .22432 ??????? "Are all immunohistochemical reagents used within their indicated dates? How are other institutions managing their antibodies that require freezing.--- do you aliquot,freeze, and give the reagent an extended expiration after it has been diluted? Thanks in advance!!! I have read the past posts on this in the archives, but am wondering if there is a clear answer to antibody use and freezing. Theresa R Dobersztyn HT (ASCP) Electron Microscopy/Histology Labs Senior Technologist Department of Pathology Akron Children's Hospital 1 Perkins Square Akron, Ohio 44308 330-543-8279 Akron Children's Hospital - Proud winner of the NorthCoast 99 "Best Workplace" award! ************************************************************************* This electronic mail transmission, including any attached files, may contain confidential and/or privileged information for the sole use of the intended recipient(s). It is not intended for transmission to, or receipt by, any unauthorized parties. Any review, use, distribution, dissemination, downloading, copying or disclosure by others is strictly prohibited. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that you have received this transmission in error. If you have received this transmission in error, please delete it, as well as any copies, from your system without copying it, and notify the sender by reply e-mail. From Rcartun <@t> harthosp.org Thu May 21 13:33:02 2009 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Thu May 21 13:33:19 2009 Subject: [Histonet] Dry ice - transportation Message-ID: <4A15661D.7400.0077.1@harthosp.org> Are there any restrictions on the transportation of dry ice (and human tissue) in private vehicles? Our PAs cover a surgical center off-site and we need to transport frozen tissue back to the hospital for biobanking purposes. They drive back and forth in their own vehicles. Thanks. Richard Richard W. Cartun, Ph.D. Director, Histology & Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From LSebree <@t> uwhealth.org Thu May 21 13:51:37 2009 From: LSebree <@t> uwhealth.org (Sebree Linda A) Date: Thu May 21 13:51:42 2009 Subject: [Histonet] IHC question- freezing aliquots In-Reply-To: Message-ID: <5998F3BDFF7AAC4091C7AE93A7A1A5892CDFF7@UWHC-MAIL01.uwhis.hosp.wisc.edu> In a similar situation we have gotten documentation (e-mail) from the vendor stating the recommendation to freeze aliquots and to what degree that extends the shelf life. We haven't been queried on these particular antibodies by an inspector but our supervisor feels that this satisfies the CAP reg. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of tdobersztyn@chmca.org Sent: Thursday, May 21, 2009 1:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC question- freezing aliquots Hello all! I have an antibody that has a "2" month expiration date. It is recommended that we aliquot, and freeze (-80) the antibody- per the vendor. How do you interpret: The CAP reg pertaining to expired antibody use: ANP .22432 ??????? "Are all immunohistochemical reagents used within their indicated dates? How are other institutions managing their antibodies that require freezing.--- do you aliquot,freeze, and give the reagent an extended expiration after it has been diluted? Thanks in advance!!! I have read the past posts on this in the archives, but am wondering if there is a clear answer to antibody use and freezing. Theresa R Dobersztyn HT (ASCP) Electron Microscopy/Histology Labs Senior Technologist Department of Pathology Akron Children's Hospital 1 Perkins Square Akron, Ohio 44308 330-543-8279 Akron Children's Hospital - Proud winner of the NorthCoast 99 "Best Workplace" award! ************************************************************************ * This electronic mail transmission, including any attached files, may contain confidential and/or privileged information for the sole use of the intended recipient(s). It is not intended for transmission to, or receipt by, any unauthorized parties. Any review, use, distribution, dissemination, downloading, copying or disclosure by others is strictly prohibited. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that you have received this transmission in error. If you have received this transmission in error, please delete it, as well as any copies, from your system without copying it, and notify the sender by reply e-mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Luis.Chiriboga <@t> nyumc.org Thu May 21 13:55:10 2009 From: Luis.Chiriboga <@t> nyumc.org (Chiriboga, Luis) Date: Thu May 21 13:55:17 2009 Subject: [Histonet] Plant/insect histology Message-ID: <602D3639FB75D7479E63338AB2E82AEA07CE46BB3B@MSGWSDCPMB05.nyumc.org> If anyone out in histonet land is doing above, could you please contact me by email directly? Thanks Luis ------------------------------------------------------------
This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email.
================================= 
From joseph-galbraith <@t> uiowa.edu  Thu May 21 14:08:46 2009
From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe)
Date: Thu May 21 14:08:56 2009
Subject: [Histonet] Dry ice - transportation
In-Reply-To: <4A15661D.7400.0077.1@harthosp.org>
References: <4A15661D.7400.0077.1@harthosp.org>
Message-ID: 

Richard:

There may be local regulations that could vary.  Here you can buy dry
ice in the grocery store (if you bring an appropriate container).  The
safety warnings associated with dry ice indicate (as you would expect) -
transport in unsealed insulated container (Styrofoam chest), transport
in the trunk of your car not the passenger compartment and crack a
window, avoid direct contact with skin eyes or ingestion to avoid burns,
etc.  If your PA's are traveling long distances using a minivan SUV or
similar single compartment vehicle, the main problem would be
accumulation of CO2 gas that would ultimately induce drowsiness and in
worst case scenario asphyxia.

Joe

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard
Cartun
Sent: Thursday, May 21, 2009 1:33 PM
To: Histonet
Subject: [Histonet] Dry ice - transportation

Are there any restrictions on the transportation of dry ice (and human
tissue) in private vehicles?  Our PAs cover a surgical center off-site
and we need to transport frozen tissue back to the hospital for
biobanking purposes.  They drive back and forth in their own vehicles.
Thanks.

Richard 

Richard W. Cartun, Ph.D.
Director, Histology & Immunopathology
Director, Biospecimens
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-0174 Fax


_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

From af46 <@t> buffalo.edu  Thu May 21 14:28:54 2009
From: af46 <@t> buffalo.edu (Annette Featherstone)
Date: Thu May 21 14:28:59 2009
Subject: [Histonet] Adhesive for immunos
Message-ID: <000001c9da4a$61318480$23948d80$@edu>

We have used 5% Tite-bond glue(  from the Hardware store) on slides with
great success. Just take a Kimwipe and wipe it evenly over the slide and it
holds the tissue very well.

 

Annette Featherstone, 

Supervisor of Pathology and Anatomical Sciences

The University of Buffalo

Phone:716-829-3108

Fax: 716-829-2911

 

From rjbuesa <@t> yahoo.com  Thu May 21 14:49:06 2009
From: rjbuesa <@t> yahoo.com (Rene J Buesa)
Date: Thu May 21 14:49:11 2009
Subject: [Histonet] cloudy formalin in tissue processor
Message-ID: <384163.89747.qm@web65712.mail.ac4.yahoo.com>

If you use NBF it will get cloudy when it mixes (in some step) with alcohol. The NBF has salts that are not soluble in alcohol and will produce cloudiness. You are getting your NBF mixed with alcohol.
Ren? J.

--- On Thu, 5/21/09, Burton, Lynn  wrote:


From: Burton, Lynn 
Subject: RE: [Histonet] cloudy formalin in tissue processor
To: "Christine I. Braaten" , histonet@lists.utsouthwestern.edu
Date: Thursday, May 21, 2009, 1:23 PM


Is your formalin picking up water from the air in the room? Check the humidity level in your room.

Lynn Burton
Lab Assoc. I
Animal Disease Lab
Galesburg, Il 61401

________________________________

From: histonet-bounces@lists.utsouthwestern.edu on behalf of Christine I. Braaten
Sent: Thu 5/21/2009 11:41 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] cloudy formalin in tissue processor



Has anyone had a problem with their formalin getting cloudy in their
tissue processor? We have a VIP tissue processor and just recently
started having problems with cloudy formalin. Any suggestions would be
appreciated. I checked the archives and couldn't find any articles on
this particular subject. Thanks.


CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients 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 electronic mail and destroy all copies of the original message.
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet


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Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



      
From mucram11 <@t> comcast.net  Thu May 21 14:57:28 2009
From: mucram11 <@t> comcast.net (Pamela Marcum)
Date: Thu May 21 14:57:31 2009
Subject: [Histonet] cloudy formalin in tissue processor
In-Reply-To: <384163.89747.qm@web65712.mail.ac4.yahoo.com>
Message-ID: <603333943.107351242935847970.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net>



This can result from not drying the retort or tissue tank after the clean cycle which will leave alcohol in the bottom.? Dry the tank beofre starting the next run.? Ocassionally I have even seen some xylene end up in a mixture in the tissue tank if it is not dried well.? Both of these will turn your NBF cloudy.?? When you get a processor they don't always tell you to dry it to prevent carry over. 



Pam Marcum 

UPENN Vet Sch 

New Bolton Center 

----- Original Message ----- 
From: "Rene J Buesa"  
To: "Christine I. Braaten" , histonet@lists.utsouthwestern.edu, "LynnBurton"  
Sent: Thursday, May 21, 2009 3:49:06 PM GMT -05:00 US/Canada Eastern 
Subject: RE: [Histonet] cloudy formalin in tissue processor 

If you use NBF it will get cloudy when it mixes (in some step) with alcohol. The NBF has salts that are not soluble in alcohol and will produce cloudiness. You are getting your NBF mixed with alcohol. 
Ren? J. 

--- On Thu, 5/21/09, Burton, Lynn  wrote: 


From: Burton, Lynn  
Subject: RE: [Histonet] cloudy formalin in tissue processor 
To: "Christine I. Braaten" , histonet@lists.utsouthwestern.edu 
Date: Thursday, May 21, 2009, 1:23 PM 


Is your formalin picking up water from the air in the room? Check the humidity level in your room. 

Lynn Burton 
Lab Assoc. I 
Animal Disease Lab 
Galesburg, Il 61401 

________________________________ 

From: histonet-bounces@lists.utsouthwestern.edu on behalf of Christine I. Braaten 
Sent: Thu 5/21/2009 11:41 AM 
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] cloudy formalin in tissue processor 



Has anyone had a problem with their formalin getting cloudy in their 
tissue processor? We have a VIP tissue processor and just recently 
started having problems with cloudy formalin. Any suggestions would be 
appreciated. I checked the archives and couldn't find any articles on 
this particular subject. Thanks. 


CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients 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 electronic mail and destroy all copies of the original message. 
_______________________________________________ 
Histonet mailing list 
Histonet@lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 


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Histonet@lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 




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From Chris.Stephenson <@t> comphealth.com  Thu May 21 15:03:32 2009
From: Chris.Stephenson <@t> comphealth.com (Chris.Stephenson@comphealth.com)
Date: Thu May 21 15:03:44 2009
Subject: [Histonet] Chris Stephenson is out of the office.
Message-ID: 


I will be out of the office starting  05/21/2009 and will not return until
05/26/2009.

Please forward all urgent messages to Eric Halpern at ext. 5852 or
Eric.Halpern@comphealth.com
From rsrichmond <@t> gmail.com  Thu May 21 15:21:17 2009
From: rsrichmond <@t> gmail.com (Robert Richmond)
Date: Thu May 21 15:21:23 2009
Subject: [Histonet] Re: Bone saw
Message-ID: 

Christine Bark asks about saws for cutting surgical pathology bone
specimens like femoral heads.

I posted something to Histonet about this fairly recently - here it is again.
****************************
Stephen Peters, a pathologist in Hackensack NJ, describes for us a
bone saw he's invented. His Web site is distinctly worth looking at,
the whole thing, not just the saw.
http://pathologyinnovations.com/bone_vise.htm

Some other possibilities for sawing bone:

Many of the small pathology services I work for have no way of sawing
bone. (It's amazing how many pathologists there are who are so poorly
trained that they think you decalcify a femoral head by tossing the
whole thing in formalin for a month.) In that circumstance, I head for
the nearest hardware store and buy a five dollar hacksaw which I leave
behind at the end of the week.

The Civil War vintage Satterlee amputation saw is still available, and
is a serviceable hand saw that doesn't go dull quickly. (I've seen
them, complete with chrome plating, at Civil War re-enactments.) This
is the tool I most commonly use.

One of the standard vendors offers a simple device for slabbing a
femoral head, the SawBones, absurdly overpriced at $500, not something
a hospital would be likely to buy for a mere pathologist.

Several years ago I attended a continuing medical education program
where the lecturer recommended a scroll saw, a large table saw that's
about impossible to injure yourself with. At the time they cost about
$100 for Made in China, otherwise $200. The disadvantage, in a cramped
pathology lab, is its large footprint. You can look at these things at
your local Home Depot.

There's no way to cut a femoral head safely with an oscillating
autopsy saw (Stryker saw), though this is probably the most common way
to cut bone.

I think femoral heads removed for fracture (not for osteoarthritis)
really do need to be examined microscopically, because of the
occasional pathologic fracture (fracture through metastatic cancer in
the bone). I've seen several of these, not all with a previous cancer
diagnosis. (But I see no reason to examine knee replacement material
microscopically, if you know how to do the gross description properly
- which admittedly most pathologists don't.)

(I have no connection with any of the businesses I've mentioned.)

Bob Richmond
Samurai Pathologist
Knoxville TN

From AnthonyH <@t> chw.edu.au  Thu May 21 17:57:00 2009
From: AnthonyH <@t> chw.edu.au (Tony Henwood)
Date: Thu May 21 17:57:14 2009
Subject: [Histonet] RE: [IHCRG] HRP-labeled primary antibodies
In-Reply-To: <905854.25089.qm@web50310.mail.re2.yahoo.com>
Message-ID: 

Kim,
 
The direct IPX is the easiest method apart from a direct
immunofluorescence. Very few steps:
1.    Block endogenous enzyme
2.    Block non-specific Ab binding
3.    Put the HRP-conjugated Ab on (a an appropriate dilution, usually
more concentrated than if you were using an ABC or Polymer amplification
method)
4.    Incubate in your usual DAB-H2O2 solution
5.    Counterstain and you done.
 
Throw in a few buffer washes in between and maybe some antigen retrieval
if required.
 
We use the direct IPX method for renal biopsies (IgG,M,A etc). See
Birchall, I.W., (1996) "Direct antibody method for formalin fixed,
paraffin embedded renal biopsies: a comparison with the peroxidase
labelled streptavidin/biotin method" J. Histotechnol 19(2): 125-129.

Regards 

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 


	-----Original Message-----
	From: ihcrg@googlegroups.com [mailto:ihcrg@googlegroups.com] On
Behalf Of Kim Merriam
	Sent: Thursday, 21 May 2009 10:42 PM
	To: Histonet; ihcrg@googlegroups.com
	Subject: [IHCRG] HRP-labeled primary antibodies
	
	
	Hi All,
	 
	Has anyone had experience doing IHC on FFPE tissues with
HRP-labeled primary antibodies?  I was wondering what the best way to
detect them would be.  I assume that going strait to DAB would not work,
since no amplification is there.  I was thinking of using a biotinyl
tyramide step to amplify the signal.
	 
	Also, do you think the final antibody concentration would need
to be higher than with traditional, unlabeled primaries?
	 
	Thanks,
	Kim
	 
	Kim Merriam, MA, HT(ASCP)QIHC
	Cambridge, MA 



*********************************************************************
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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

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From AnthonyH <@t> chw.edu.au  Thu May 21 18:00:38 2009
From: AnthonyH <@t> chw.edu.au (Tony Henwood)
Date: Thu May 21 18:00:46 2009
Subject: [Histonet] RE: [IHCRG] Re: CMV antibody
In-Reply-To: 
Message-ID: 

Interestingly,

This data sheet does not seem to list the clone of the CMV monoclonal
antibody, or am I misreading it?


Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-----Original Message-----
From: ihcrg@googlegroups.com [mailto:ihcrg@googlegroups.com] On Behalf
Of michael.ho@sickkids.ca
Sent: Thursday, 21 May 2009 5:20 AM
To: JTAYLOR@meriter.com
Cc: 'histonet@lists.utsouthwestern.edu'; 'ihcrg@googlegroups.com'
Subject: [IHCRG] Re: CMV antibody


See attached data sheet, we use pepsin  enzyme digestion @ 37C for 30
minutes manually on FFPE

For Ventana Users Protease I for 16 minutes





Michael  Ho, MLT



Resource Technologist(See attached file: CMV-E.pdf) Immunopathology
laboratory Division of Pathology DPLM The Hospital for Sick Children
Toronto, Ontario Canada Tel#:416-813-5950
Fax: 416-813-5974


 

             "Taylor, Jean"

             
To 
             Sent by:                  "'ihcrg@googlegroups.com'"

             ihcrg@googlegroup         ,

             s.com
"'histonet@lists.utsouthwestern.edu 
                                       '"

 
 
             2009-05-20 03:06
cc 
             PM

 
Subject 
                                       [IHCRG] CMV antibody

             Please respond to

             JTAYLOR@meriter.c

                    om

 

 

 





Hi Everyone,

I'm wondering what antibody clone labs are using to detect
Cytomegalovirus by IHC.

Thank you!

Jean Taylor, HT(ASCP) QIHC
IHC Tech.
Meriter Health Services
Madison, WI 53715

*********************************************************************
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 note also confirms that this email message has been
virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses.
**********************************************************************


From AnthonyH <@t> chw.edu.au  Thu May 21 18:03:00 2009
From: AnthonyH <@t> chw.edu.au (Tony Henwood)
Date: Thu May 21 18:03:08 2009
Subject: [Histonet] cloudy formalin in tissue processor
In-Reply-To: 
Message-ID: 

Possible carry over of xylene from the flush program?

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Christine I. Braaten
Sent: Friday, 22 May 2009 2:42 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] cloudy formalin in tissue processor


Has anyone had a problem with their formalin getting cloudy in their
tissue processor? We have a VIP tissue processor and just recently
started having problems with cloudy formalin. Any suggestions would be
appreciated. I checked the archives and couldn't find any articles on
this particular subject. Thanks. 


CONFIDENTIALITY NOTICE: This electronic message, including any
attachments, is for the sole use of the intended recipients 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 electronic mail and
destroy all copies of the original message.
_______________________________________________
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 note also confirms that this email message has been
virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses.
**********************************************************************


From AnthonyH <@t> chw.edu.au  Thu May 21 18:28:11 2009
From: AnthonyH <@t> chw.edu.au (Tony Henwood)
Date: Thu May 21 18:28:16 2009
Subject: [Histonet] RE: [IHCRG] Re: CMV antibody
In-Reply-To: 
Message-ID: 

Michael,

I have used both the Dako (Clones CCH2 + DDG9) and the Novocastra
products (Clones 2 and 6) and they both stain the same.

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-----Original Message-----
From: michael.ho@sickkids.ca [mailto:michael.ho@sickkids.ca] 
Sent: Friday, 22 May 2009 9:22 AM
To: Tony Henwood
Cc: histonet@lists.utsouthwestern.edu; ihcrg@googlegroups.com;
JTAYLOR@meriter.com
Subject: Re: [IHCRG] Re: CMV antibody


Hi Tony
If you are interested I can email more information...touch base with me
....what are you using at your institute???

best wishes


Michael  Ho, MLT
Resource Technologist
Immunopathology laboratory
Division of Pathology
DPLM
The Hospital for Sick Children
Toronto, Ontario
Canada
Tel#:416-813-5950
Fax: 416-813-5974


 

             "Tony Henwood"

             
To 
             Sent by:                  ,

             ihcrg@googlegroup         

             s.com
cc 
 
 
                                       , 

             05/21/2009 07:00
Subject 
             PM                        [IHCRG] Re: CMV antibody

 

 

             Please respond to

             anthonyh@chw.edu.

                    au

 

 






Interestingly,

This data sheet does not seem to list the clone of the CMV monoclonal
antibody, or am I misreading it?


Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory
Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA




-----Original Message-----
From: ihcrg@googlegroups.com [mailto:ihcrg@googlegroups.com] On Behalf
Of michael.ho@sickkids.ca
Sent: Thursday, 21 May 2009 5:20 AM
To: JTAYLOR@meriter.com
Cc: 'histonet@lists.utsouthwestern.edu'; 'ihcrg@googlegroups.com'
Subject: [IHCRG] Re: CMV antibody


See attached data sheet, we use pepsin  enzyme digestion @ 37C for 30
minutes manually on FFPE

For Ventana Users Protease I for 16 minutes





Michael  Ho, MLT



Resource Technologist(See attached file: CMV-E.pdf) Immunopathology
laboratory Division of Pathology DPLM The Hospital for Sick Children
Toronto, Ontario Canada Tel#:416-813-5950
Fax: 416-813-5974




             "Taylor, Jean"

             
To
             Sent by:                  "'ihcrg@googlegroups.com'"

             ihcrg@googlegroup         ,

             s.com
"'histonet@lists.utsouthwestern.edu
                                       '"



             2009-05-20 03:06
cc
             PM


Subject
                                       [IHCRG] CMV antibody

             Please respond to

             JTAYLOR@meriter.c

                    om











Hi Everyone,

I'm wondering what antibody clone labs are using to detect
Cytomegalovirus by IHC.

Thank you!

Jean Taylor, HT(ASCP) QIHC
IHC Tech.
Meriter Health Services
Madison, WI 53715

*********************************************************************
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 note 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.
**********************************************************************




*********************************************************************
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 note 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 matthewtclose <@t> gmail.com  Thu May 21 19:57:29 2009
From: matthewtclose <@t> gmail.com (Matthew Close)
Date: Thu May 21 19:57:38 2009
Subject: [Histonet] re:cloudy formalin
Message-ID: <6abc767b0905211757j3bdba310n9dd9354b15910e92@mail.gmail.com>

Are you certain that it is formalin and not paraformaldehyde?  The latter
will get cloudy.  Also, is there any way that your clearing solvent might be
making its way in to the formalin solution?
From kwuny <@t> email.cs.nsw.gov.au  Thu May 21 22:17:37 2009
From: kwuny <@t> email.cs.nsw.gov.au (Young Kwun)
Date: Thu May 21 22:17:46 2009
Subject: [Histonet] Antibodies for Alport syndrome
Message-ID: <001b01c9da8b$dbfc3ca0$c680310a@cs.nsw.gov.au>

 

Hi All,

Has anyone had experience doing IHC on FFPE tissues with Collagen Type IV
subunit antibodies?

These are Collagen Type IV alpha 1, alpha 3 and alpha 5 chains.  They are
used for the diagnosis of Alport syndrome and all antibodies are from KAMIYA
BIOMEDICAL COMPANY, Seatle. At present, it seems that only COL4A5 is working
on FFPE tissues.

I tried with (or without) various pre-treatment before the primary
incubation with the dilution of 1:50 to 1:500.

I am using BondMax autostainer and I also tried Decloaking chamber HIER with
manual staining.

I was wondering whether there would be other source for these antibodies.

Thank you. 

 

Regards,

 

Young

 

 

 

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

 

 

From Allison_Scott <@t> hchd.tmc.edu  Thu May 21 22:37:45 2009
From: Allison_Scott <@t> hchd.tmc.edu (Scott, Allison D)
Date: Thu May 21 22:37:57 2009
Subject: [Histonet] Wholemount processing
Message-ID: <1872B4A455B7974391609AD8034C79FC019C18AA@LBEXCH01.hchd.local>

Hello to all in histoland.  My lab is looking to start processing whole prostate specimens and doing the wholemount
process.  Can anyone give me a starting point on how to proceed.  I would need information on processing times,
embedding and cutting equipment, staining information and the glass that you mount the specimens on.  Any help in this matter would be greatly appreciated.
 
Allison Scott HT(ASCP)
Histology Supervisor
LBJ Hospital
Houston, Texas 77026
CONFIDENTIALITY NOTICE:
If you have received this e-mail in error, please immediately notify the
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From Kemlo.Rogerson <@t> waht.swest.nhs.uk  Fri May 22 02:21:07 2009
From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson)
Date: Fri May 22 02:21:25 2009
Subject: [Histonet] cloudy formalin in tissue processor
Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06C0D2CC@wahtntex2.waht.swest.nhs.uk>

The old tissue processors used to have cloudy formalin to the point
you'd get a layer at the bottom of the jars. If it is not contamination
from the clearing fluids already suggested, could it not just be the
fixed protein from the tissues that is held in suspension. I would have
thought very bloody or very proteinaseous tissue would have these labile
proteins fixed which would then fall out of solution; from memory the
cold makes it worse.


Kemlo Rogerson	
e-mail kemlorogerson@nhs.net if not at work.			
DD   01934 647057 or extension 3311     Mob 07749 754194; 
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer
This e-mail is confidential and privileged. If you are not the intended
recipient please accept my apologies; please do not disclose, copy or
distribute information in this e-mail or take any action in reliance on
its contents: to do so is strictly prohibited and may be unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation

 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Christine I. Braaten
Sent: 21 May 2009 17:42
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] cloudy formalin in tissue processor

Has anyone had a problem with their formalin getting cloudy in their
tissue processor? We have a VIP tissue processor and just recently
started having problems with cloudy formalin. Any suggestions would be
appreciated. I checked the archives and couldn't find any articles on
this particular subject. Thanks. 


CONFIDENTIALITY NOTICE: This electronic message, including any
attachments, is for the sole use of the intended recipients 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 electronic mail and
destroy all copies of the original message.
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

From Kemlo.Rogerson <@t> waht.swest.nhs.uk  Fri May 22 02:24:49 2009
From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson)
Date: Fri May 22 02:25:01 2009
Subject: [Histonet] Bone!
Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06C0D2CE@wahtntex2.waht.swest.nhs.uk>

Probably of all the fixatives 10% neutral buffered formalin is least
likely to overfix and I've left tissues in it for years. Formalin, as
you know, is one of those fixatives that you can wash out. There are
preservatives, rather than fixatives, you could try but I don't have
experience of those; my best advise is formalin but make sure that it is
neutral and buffered. I think the effects of pH would be more damaging
than the relatively 'soft' formalin fixation.



Kemlo Rogerson	
e-mail kemlorogerson@nhs.net if not at work.			
DD   01934 647057 or extension 3311     Mob 07749 754194; 
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer
This e-mail is confidential and privileged. If you are not the intended
recipient please accept my apologies; please do not disclose, copy or
distribute information in this e-mail or take any action in reliance on
its contents: to do so is strictly prohibited and may be unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation

 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert
Edward Pogue
Sent: 21 May 2009 16:33
To: Kemlo Rogerson
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Bone!

Thanks Kemlo...

I thought of that, but was worried about over-fixation,- what do you
think?

Redward.


------------------------------
 
In 10% neutral buffered formalin?



Kemlo Rogerson	
e-mail kemlorogerson@nhs.net if not at work.			
DD   01934 647057 or extension 3311     Mob 07749 754194; 
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer This e-mail is confidential and privileged. If you are not the
intended recipient please accept my apologies; please do not disclose,
copy or distribute information in this e-mail or take any action in
reliance on its contents: to do so is strictly prohibited and may be
unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation

 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert
Edward Pogue
Sent: 21 May 2009 14:47
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Bone!

Friends,

Can someone recommend to me how to store bones (juvenile rat femur) that
I have fixed and decalcified, but am not yet ready to cut (they're for
vibratome, so not embedded).

Thanks!

Redward.

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From Barry.R.Rittman <@t> uth.tmc.edu  Fri May 22 07:20:38 2009
From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R)
Date: Fri May 22 07:20:42 2009
Subject: [Histonet] Bone!
In-Reply-To: <86ADE4EB583CE64799A9924684A0FBBF06C0D2CE@wahtntex2.waht.swest.nhs.uk>
References: <86ADE4EB583CE64799A9924684A0FBBF06C0D2CE@wahtntex2.waht.swest.nhs.uk>
Message-ID: <75A0543E23D3A7458012D9E02EDBEC0002E927278C@UTHCMS1.uthouston.edu>

Lets face it there is no really ideal medium for long term storage.
However this depends on what techniques you want to carry out later and when.
I agree that 10% NBF is a good fixative for bone, I would definitely not advise leaving it in this fixative for extended periods of time.
The reason is that formalin continues cross linking.
This means that many stains and immunohistochemistry may no longer be possible depending on the amount of time left in fixative.
Also Kemlo is correct in that temporary formalin bonds can be broken by extended washing in watre. This only applies to the weak temporary bonds and not to the more stable bonds. The longer the fixing the greater the number of stable bonds.
I would recommend having a mixture of 70% ethanol plus glycerin up to 10-20%.  The glycerin will prevent any drying out of the solution over extended periods of tim.
Barry


________________________________________
From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kemlo Rogerson [Kemlo.Rogerson@waht.swest.nhs.uk]
Sent: Friday, May 22, 2009 2:24 AM
To: Robert Edward Pogue
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Bone!

Probably of all the fixatives 10% neutral buffered formalin is least
likely to overfix and I've left tissues in it for years. Formalin, as
you know, is one of those fixatives that you can wash out. There are
preservatives, rather than fixatives, you could try but I don't have
experience of those; my best advise is formalin but make sure that it is
neutral and buffered. I think the effects of pH would be more damaging
than the relatively 'soft' formalin fixation.



Kemlo Rogerson
e-mail kemlorogerson@nhs.net if not at work.
DD   01934 647057 or extension 3311     Mob 07749 754194;
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-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert
Edward Pogue
Sent: 21 May 2009 16:33
To: Kemlo Rogerson
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Bone!

Thanks Kemlo...

I thought of that, but was worried about over-fixation,- what do you
think?

Redward.


------------------------------

In 10% neutral buffered formalin?



Kemlo Rogerson
e-mail kemlorogerson@nhs.net if not at work.
DD   01934 647057 or extension 3311     Mob 07749 754194;
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer This e-mail is confidential and privileged. If you are not the
intended recipient please accept my apologies; please do not disclose,
copy or distribute information in this e-mail or take any action in
reliance on its contents: to do so is strictly prohibited and may be
unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation



-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert
Edward Pogue
Sent: 21 May 2009 14:47
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Bone!

Friends,

Can someone recommend to me how to store bones (juvenile rat femur) that
I have fixed and decalcified, but am not yet ready to cut (they're for
vibratome, so not embedded).

Thanks!

Redward.

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From pkarlisch <@t> hmc.psu.edu  Fri May 22 08:20:53 2009
From: pkarlisch <@t> hmc.psu.edu (Patricia Karlisch)
Date: Fri May 22 08:21:28 2009
Subject: [Histonet] RE: Dry Ice Transportation
Message-ID: <4A166E74.07B7.008C.0@hmc.psu.edu>

I think you may need to check the regs of your DOH.  We did this in a neighboring state for a short period but were advised by the distributors that this can be a health hazard in a closed vehicle depending on the amount of dry ice. Pat

Pat Karlisch
Supervisor, Histology, Pathology and Laboratory Medicine
Penn State Milton S. Hershey Medical Center
Mail Code H179
Hershey, PA 17033
Phone (717) 531-6072
Fax: (717) 531- 7741
email: pkarlisch@psu.edu

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From loftonjt <@t> holycrosshealth.org  Fri May 22 08:34:28 2009
From: loftonjt <@t> holycrosshealth.org (Jimmy Lofton)
Date: Fri May 22 08:34:48 2009
Subject: [Histonet] Bone Marrow Clot
Message-ID: <4A1671A4.9B70.0056.0@holycrosshealth.org>

Has anyone had problems with bone marrow clots where the inside is well preserved and the cells around the rim are staining lighter and some rbc's may be lyced.  We use the thrombin clot method.
 
 
Thanks,
 
Jimmy
 
 
Jimmy Lofton, M.S., HT,CT(ASCP)
Manager Histology Laboratory
Holy Cross Hospital
1500 Forest Glen Road
Silver Spring, MD  20910-1484
301-754-7353 (Phone)
301-754-8563 (Fax)
loftonjt@holycrosshealth.org
 

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From grudow1 <@t> jhmi.edu  Fri May 22 08:38:15 2009
From: grudow1 <@t> jhmi.edu (Gay Rudow)
Date: Fri May 22 08:38:19 2009
Subject: [Histonet] microtome and processing for large specimens
Message-ID: <864A73846CE8F04D995603351682C29C871CE70B56@JHEMTEXVS2.win.ad.jhu.edu>

Hi,

We are going to start on a new project in which we will need to cut a whole paraffin-embedded brain hemisphere. Does anyone know of a company that carries a microtome made for cutting large sections? Also, if anyone has suggestions as to how to embed very large sections or how many pieces I will need to cut the brain into in order to accomplish this. Thanks for your help!
Gay
From POWELL_SA <@t> mercer.edu  Fri May 22 09:05:55 2009
From: POWELL_SA <@t> mercer.edu (Shirley A. Powell)
Date: Fri May 22 09:06:32 2009
Subject: [Histonet] RE: microtome and processing for large specimens
In-Reply-To: <864A73846CE8F04D995603351682C29C871CE70B56@JHEMTEXVS2.win.ad.jhu.edu>
References: <864A73846CE8F04D995603351682C29C871CE70B56@JHEMTEXVS2.win.ad.jhu.edu>
Message-ID: <9BF995BC0E47744E9673A41486E24EE21AB0D135D5@MERCERMAIL.MercerU.local>

Hi Gay, 
I can help you with your project.  I am doing a workshop (#40) at NSH this year on sectioning macros as well if you are going to be there.  Are you cutting human or animal, the size is important.  Some animal brains can be sectioned on regular rotary microtomes if there is enough blade clearance.  

To answer your question about microtomes Leica carries sliding microtomes which can handle large blocks/whole organ blocks.  Give me a call at 478-301-2374.  

Shirley Powell
Mercer University School of Medicine
Macon, Georgia

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gay Rudow
Sent: Friday, May 22, 2009 9:38 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] microtome and processing for large specimens

Hi,

We are going to start on a new project in which we will need to cut a whole paraffin-embedded brain hemisphere. Does anyone know of a company that carries a microtome made for cutting large sections? Also, if anyone has suggestions as to how to embed very large sections or how many pieces I will need to cut the brain into in order to accomplish this. Thanks for your help!
Gay
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From POWELL_SA <@t> mercer.edu  Fri May 22 09:17:27 2009
From: POWELL_SA <@t> mercer.edu (Shirley A. Powell)
Date: Fri May 22 09:18:02 2009
Subject: [Histonet] RE: Whole mount processing
In-Reply-To: <1872B4A455B7974391609AD8034C79FC019C18AA@LBEXCH01.hchd.local>
References: <1872B4A455B7974391609AD8034C79FC019C18AA@LBEXCH01.hchd.local>
Message-ID: <9BF995BC0E47744E9673A41486E24EE21AB0D135F0@MERCERMAIL.MercerU.local>

Hi Allison, 

I just responded to a similar request.  I can help you.  I do whole mount sections, macros, and will be presenting a workshop at NSH (#40) in Birmingham AL in October.  Whole prostates were done at the hospital where I used to work and we were able to section them on the regular rotary Leica microtomes and mounting on 2x2 slides.  If larger they were sectioned on a sliding microtome and mounted on 3.25" x 4" slides.  For larger organs, brain, heart, lung, I use 5" x 7" slides.  Give me a call at 478-301-2374.  

Shirley Powell

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Scott, Allison D
Sent: Thursday, May 21, 2009 11:38 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Wholemount processing

Hello to all in histoland.  My lab is looking to start processing whole prostate specimens and doing the wholemount
process.  Can anyone give me a starting point on how to proceed.  I would need information on processing times,
embedding and cutting equipment, staining information and the glass that you mount the specimens on.  Any help in this matter would be greatly appreciated.
 
Allison Scott HT(ASCP)
Histology Supervisor
LBJ Hospital
Houston, Texas 77026
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From tifei <@t> foxmail.com  Fri May 22 10:28:27 2009
From: tifei <@t> foxmail.com (TF)
Date: Fri May 22 10:28:51 2009
Subject: [Histonet] Preparation and store of the 1mM CaCl2 in PBS
Message-ID: <200905222328221911506@foxmail.com>

Hi all

I am using 1mM Ca2+ PBS and I found the soultion become cloudy within 6 hours after preparation...
Should because of CO2 entering into the solution?

I wonder anyone has a suggestion in using this kind of solution? I dont want to prepare it everytime...
Thanks very much!

2009-05-22 



TF 
From gagnone <@t> KGH.KARI.NET  Fri May 22 11:29:16 2009
From: gagnone <@t> KGH.KARI.NET (Gagnon, Eric)
Date: Fri May 22 11:29:25 2009
Subject: [Histonet] Prostate Whole Mounts
Message-ID: 

Hi Allison,
We cut our prostate whole mounts on the Leica RM2255 automated microtome.  We obtained their Super Mega Cassette Clamp, (part no 140502 38967).  This clamp mounts easily onto the microtome, and holds SurgiPath SuperCassettes ( Cat No VSP59067B-BX grey in colour).  The beauty of this microtome is that the stroke is 10 mm longer than our previous Reichert-Jung 2035 microtomes we used to cut these on, I believe it is now 70 mm vertically.  In terms of slides, we use 75x38 up to 75x80 mm slides, with coverslips ranging from 35x50 to 48x65 mm, depending on specimen size.  The slides go on our Leica automated stainer, held diagonally in the basket by other slides to keep them vertical, and coverslipped by hand.

In terms of processing, we use our normal overnight processing cycle as we would for our routine surgical blocks.  These sections are well-fixed before processing, cut at 4-5 mm during crossing.  We have had some issues when the pathologist assistants couldn't get the sections this thin, but it hasn't happened often enough to use a longer/different processing cycle.  We have been getting 4-6 slides blocks per case, and we've done 35 cases in the last year. 
 
Hope this helps,
Eric Gagnon MLT
Histology Laboratory
Kingston General Hospital
Kingston, Ontario, Canada
 

 
 


From CBraaten <@t> Cheshire-Med.COM  Fri May 22 11:36:13 2009
From: CBraaten <@t> Cheshire-Med.COM (Christine I. Braaten)
Date: Fri May 22 11:37:40 2009
Subject: [Histonet] cloudy formalin in vip tissue processor
Message-ID: 

Thanks to all who have replied to my query. I probably should have
mentioned that we use zinc formalin in our processor and as our primary
fixative. I have spoken to both Sakura and Anatech (supplier of zinc
formalin) and feel more confused. Anatech says it's a change in pH that
causes the zinc to precipitate and that xylene will cause a whole lot
more problems than a little cloudiness. Sakura thinks the zinc formalin
is the problem. We clean the retort with the cleaning program every
morning and make sure to dry the retort so I'm not sure how xylene would
get into the formalin. If this brings up any other issues I would
appreciate more feedback. Thanks 


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From rjbuesa <@t> yahoo.com  Fri May 22 12:00:02 2009
From: rjbuesa <@t> yahoo.com (rjbuesa@yahoo.com)
Date: Fri May 22 12:00:07 2009
Subject: [Histonet] Prostate Whole Mounts
Message-ID: <651.87185.qm@web65712.mail.ac4.yahoo.com>


Allison:
Eric's description of how he handles whole prostates describes almost exactly what we used to do starting in 1990.
The only thing I will add is that the?prostates should fist be opened in half and fixed for 12 hours before slicing it into further pieces. Once all the pieces are made and individually cassetted, we used to leave them in NBF formalin overnight and placed in the tissue?processor at 2PM the next day to be embedded/sectioned the next day.
?
When?we started?the mega cassettes did not exist so I used?round small plastic containers cut and will holes perforated. The embedding?molds did not exist either so I?made mine from 0.2 mm aluminum pieces?and the blocks were fixed on wood blocks like in the "good old days". At the beginning we also used a Leica vertical microtome, but we ended buying a Leica horizontal microtome.
Ren??J.?

--- On Fri, 5/22/09, Gagnon, Eric  wrote:


From: Gagnon, Eric 
Subject: [Histonet] Prostate Whole Mounts
To: histonet@lists.utsouthwestern.edu
Date: Friday, May 22, 2009, 12:29 PM


Hi Allison,
We cut our prostate whole mounts on the Leica RM2255 automated microtome.? We obtained their Super Mega Cassette Clamp, (part no 140502 38967).? This clamp mounts easily onto the microtome, and holds SurgiPath SuperCassettes ( Cat No VSP59067B-BX grey in colour).? The beauty of this microtome is that the stroke is 10 mm longer than our previous Reichert-Jung 2035 microtomes we used to cut these on, I believe it is now 70 mm vertically.? In terms of slides, we use 75x38 up to 75x80 mm slides, with coverslips ranging from 35x50 to 48x65 mm, depending on specimen size.? The slides go on our Leica automated stainer, held diagonally in the basket by other slides to keep them vertical, and coverslipped by hand.

In terms of processing, we use our normal overnight processing cycle as we would for our routine surgical blocks.? These sections are well-fixed before processing, cut at 4-5 mm during crossing.? We have had some issues when the pathologist assistants couldn't get the sections this thin, but it hasn't happened often enough to use a longer/different processing cycle.? We have been getting 4-6 slides blocks per case, and we've done 35 cases in the last year. 

Hope this helps,
Eric Gagnon MLT
Histology Laboratory
Kingston General Hospital
Kingston, Ontario, Canada






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From jcampbell <@t> vdxpathology.com  Fri May 22 12:14:27 2009
From: jcampbell <@t> vdxpathology.com (Jennifer Campbell)
Date: Fri May 22 12:14:38 2009
Subject: [Histonet] decal solution
Message-ID: <5658CBDB9EAE6545ABE50D2563D81BF821D233@VDXSERVER01.vdxpathology.local>

Does anyone know what the optimal pH for a nitric acid decal solution
should be?  Right now our's is reading below 1.0 when tested and we are
having problems with our tissue being overdecalcified.  Thank you in
advance.
 
Jennifer C.
From MElliott <@t> mrl.ubc.ca  Fri May 22 12:22:14 2009
From: MElliott <@t> mrl.ubc.ca (Mark Elliott)
Date: Fri May 22 12:22:48 2009
Subject: [Histonet] CD8 or CD4 non Mouse primary
Message-ID: <4A167CD6020000D60003F37F@mail.mrl.ubc.ca>

Hi everyone.

We are needing to do a triple stain for IF using MHC Class II and CD4 and CD8.  I have Mouse monoclonals to CD4 and CD8 and we have a goat anti-MHC.  We were wondering if anyone knows of a good CD4 or CD 8 that is made in another species that we could use to avoid using two mouse antibodies. We are doing this on human lung.

Thanks

Mark


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From leiker <@t> buffalo.edu  Fri May 22 12:33:41 2009
From: leiker <@t> buffalo.edu (Merced M Leiker)
Date: Fri May 22 12:33:48 2009
Subject: [Histonet] Determining MHC Haplotype in Pigs
Message-ID: <6209EC466929DB6E068577D1@CDYwxp1931.ad.med.buffalo.edu>

I think I will re-phrase an earlier post of mine from yeserday inquiring 
immunophenotyping in pigs.  :-) Would I would actually like to know is how 
to determine the MHC haplotype of a pig. (Their MHC is called SLA). 
Preferably using immunohistochemistry or flow cytometry.

What we ultimately would like to do is grow certain pig cells in vitro and 
then inject them back into any pig of the same haplotype without risk of 
immune rejection.

Any help or any pointers to a reference would be helpful! Thank you and 
have a nice long weekend for those celebrating the holiday....

Merced M Leiker
Cardiovascular Medicine
348 Biomedical Research Building
State University of New York at Buffalo
3435 Main St, Buffalo, NY  14214
leiker@buffalo.edu
716-829-6118

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From rgrow <@t> bmnet.com  Fri May 22 14:11:43 2009
From: rgrow <@t> bmnet.com (rgrow@bmnet.com)
Date: Fri May 22 14:11:47 2009
Subject: [Histonet] Renee Grow is out of the office.
Message-ID: 

I will be out of the office starting  05/22/2009 and will not return until
06/01/2009.

I will respond to your message when I return.
________________________________

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From rjbuesa <@t> yahoo.com  Fri May 22 15:05:13 2009
From: rjbuesa <@t> yahoo.com (Rene J Buesa)
Date: Fri May 22 15:05:18 2009
Subject: [Histonet] decal solution
Message-ID: <957365.27332.qm@web65712.mail.ac4.yahoo.com>

1- with nitric acid at high concentration as is used to decalcify, pH will be below 1
2- you cannot "overdecalcify" a tissue, but you can affect its?staining characteristics after a harsh decalcification.
Ren? J.

--- On Fri, 5/22/09, Jennifer Campbell  wrote:


From: Jennifer Campbell 
Subject: [Histonet] decal solution
To: histonet@lists.utsouthwestern.edu
Date: Friday, May 22, 2009, 1:14 PM


Does anyone know what the optimal pH for a nitric acid decal solution
should be?? Right now our's is reading below 1.0 when tested and we are
having problems with our tissue being overdecalcified.? Thank you in
advance.

Jennifer C.
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From JMacDonald <@t> mtsac.edu  Fri May 22 15:52:03 2009
From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald)
Date: Fri May 22 15:52:08 2009
Subject: [Histonet] cloudy formalin in vip tissue processor
In-Reply-To: 
Message-ID: 

Christine,
Is the zinc formalin on the tissue processor or is the tissue fixed in 
zinc formalin and then put on the tissue processor with regular 10%NBF in 
the stations?
Jennifer




"Christine I. Braaten"  
Sent by: histonet-bounces@lists.utsouthwestern.edu
05/22/2009 09:39 AM

To

cc

Subject
[Histonet] cloudy formalin in vip tissue processor






Thanks to all who have replied to my query. I probably should have
mentioned that we use zinc formalin in our processor and as our primary
fixative. I have spoken to both Sakura and Anatech (supplier of zinc
formalin) and feel more confused. Anatech says it's a change in pH that
causes the zinc to precipitate and that xylene will cause a whole lot
more problems than a little cloudiness. Sakura thinks the zinc formalin
is the problem. We clean the retort with the cleaning program every
morning and make sure to dry the retort so I'm not sure how xylene would
get into the formalin. If this brings up any other issues I would
appreciate more feedback. Thanks 


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attachments, is for the sole use of the intended recipients 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 electronic mail and 
destroy all copies of the original message.
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From Kadhem.Alkhenaizi <@t> mymail.barry.edu  Wed May 20 19:27:46 2009
From: Kadhem.Alkhenaizi <@t> mymail.barry.edu (Alkhenaizi,  Kadhem (Student))
Date: Fri May 22 16:39:33 2009
Subject: [Histonet] RE: [IHCRG] Re: CMV
In-Reply-To: 
References: <466B666475DE6547BBB0641E540A4BB5048889AE2F@EXVS1.meriter.com>,
	
Message-ID: 

We use Dako in our lab, too, with PK pretreatment.

Kadhem
HTL (ASCP) QIHC
Vitro Molecular laboratories
________________________________
From: ihcrg@googlegroups.com [ihcrg@googlegroups.com] on behalf of Tony Henwood [AnthonyH@chw.edu.au]
Sent: Wednesday, May 20, 2009 4:12 PM
To: JTAYLOR@Meriter.com; ihcrg@googlegroups.com; histonet@lists.utsouthwestern.edu
Subject: [IHCRG] Re: CMV

Jean,
No the virus is the same for IPX purposes.
I have used both the Dako (Clones CCH2 + DDG9) and the Novocastra products (Clones 2 and 6) and they both stain the same

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

-----Original Message-----
From: ihcrg@googlegroups.com [mailto:ihcrg@googlegroups.com] On Behalf Of Taylor, Jean
Sent: Thursday, 21 May 2009 6:34 AM
To: 'ihcrg@googlegroups.com'; 'histonet@lists.utsouthwestern.edu'
Subject: [IHCRG] CMV

Hi Everyone,

The pathologist that I work with wanted me to add that the CMV that we?re staining for is in the GI tract, and he?s wondering if there is a different clone/procedure that labs use for this?

Thanks again!!

Jean Taylor, HT(ASCP) QIHC
IHC Tech
Meriter Health Services
Madison, WI 53715

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From michael.ho <@t> sickkids.ca  Wed May 20 14:19:54 2009
From: michael.ho <@t> sickkids.ca (michael.ho@sickkids.ca)
Date: Fri May 22 16:39:36 2009
Subject: [Histonet] Re: [IHCRG] CMV antibody
In-Reply-To: <466B666475DE6547BBB0641E540A4BB5048889AE2E@EXVS1.meriter.com>
Message-ID: 

See attached data sheet, we use pepsin  enzyme digestion @ 37C for 30
minutes manually on FFPE

For Ventana Users Protease I for 16 minutes





Michael  Ho, MLT



Resource Technologist(See attached file: CMV-E.pdf)
Immunopathology laboratory
Division of Pathology
DPLM
The Hospital for Sick Children
Toronto, Ontario
Canada
Tel#:416-813-5950
Fax: 416-813-5974


                                                                           
             "Taylor, Jean"                                                
                                                                    To 
             Sent by:                  "'ihcrg@googlegroups.com'"          
             ihcrg@googlegroup         ,           
             s.com                     "'histonet@lists.utsouthwestern.edu 
                                       '"                                  
                                        
             2009-05-20 03:06                                           cc 
             PM                                                            
                                                                   Subject 
                                       [IHCRG] CMV antibody                
             Please respond to                                             
             JTAYLOR@meriter.c                                             
                    om                                                     
                                                                           
                                                                           
                                                                           




Hi Everyone,

I?m wondering what antibody clone labs are using to detect Cytomegalovirus
by IHC.

Thank you!

Jean Taylor, HT(ASCP) QIHC
IHC Tech.
Meriter Health Services
Madison, WI 53715
From michael.ho <@t> sickkids.ca  Wed May 20 17:43:51 2009
From: michael.ho <@t> sickkids.ca (michael.ho@sickkids.ca)
Date: Fri May 22 16:39:39 2009
Subject: [Histonet] Re: [IHCRG] CMV
In-Reply-To: <466B666475DE6547BBB0641E540A4BB5048889AE2F@EXVS1.meriter.com>
Message-ID: 

No, only need to know if you are looking for early , intermediate or late
protein expression...........we look for early to intermediate expression
at this institute...our method seems to correlate with our ISH (DNA)
method.

regards


Michael  Ho, MLT
Resource Technologist
Immunopathology laboratory
Division of Pathology
DPLM
The Hospital for Sick Children
Toronto, Ontario
Canada
Tel#:416-813-5950
Fax: 416-813-5974


                                                                           
             "Taylor, Jean"                                                
                                                                    To 
             Sent by:                  "'ihcrg@googlegroups.com'"          
             ihcrg@googlegroup         ,           
             s.com                     "'histonet@lists.utsouthwestern.edu 
                                       '"                                  
                                        
             05/20/2009 04:33                                           cc 
             PM                                                            
                                                                   Subject 
                                       [IHCRG] CMV                         
             Please respond to                                             
             JTAYLOR@meriter.c                                             
                    om                                                     
                                                                           
                                                                           
                                                                           




Hi Everyone,

The pathologist that I work with wanted me to add that the CMV that we?re
staining for is in the GI tract, and he?s wondering if there is a different
clone/procedure that labs use for this?

Thanks again!!

Jean Taylor, HT(ASCP) QIHC
IHC Tech
Meriter Health Services
Madison, WI 53715
From michael.ho <@t> sickkids.ca  Wed May 20 18:06:48 2009
From: michael.ho <@t> sickkids.ca (michael.ho@sickkids.ca)
Date: Fri May 22 16:39:42 2009
Subject: [Histonet] Re: [IHCRG] Semiautomated Antigen Retrival
	systems/equipment
In-Reply-To: 
Message-ID: 

Hi Jo ....I do not usemicrowave,  these days, even if its done in the
pressure cooker, although historically that was my initial method of
HIER(many years ago), and it worked very consistently for me, its all based
on how you standardize your method....
I moved over to the Biocare pressure cooker 5 years ago for any manual
applications I need to do, and again it has worked very well......
The bulk of my work is done on  the Ventana Benchmark and HIER online, no
problems at all, its certainly less labor intensive than any system I know
of  as it also dewaxes prior to HIER, so once you load your slides its
"Plug and Play".....Very consistent results!!!!!

I think the bottom line is go with a system you feel comfortable with,
optimize your HIER methods so that it gives you the best  results with no
tissue damage(Thats so critical)......

 hope this helps




Michael  Ho, MLT
Resource Technologist
Immunopathology laboratory
Division of Pathology
DPLM
The Hospital for Sick Children
Toronto, Ontario
Canada
Tel#:416-813-5950
Fax: 416-813-5974


                                                                           
             "Galbraith, Joe"                                              
                                                             To 
             Sent by:                  ,           
             ihcrg@googlegroup          
             s.com                                                      cc 
                                                                           
                                                                   Subject 
             05/20/2009 06:32          [IHCRG] Semiautomated Antigen       
             PM                        Retrival systems/equipment          
                                                                           
                                                                           
             Please respond to                                             
             joseph-galbraith@                                             
                 uiowa.edu                                                 
                                                                           
                                                                           




Hello all:


Is anyone using any standalone semi-automated antigen retrieval equipment?
I?ve seen a few microwave or wet bath based systems (sometimes integrated
into full blown IHC staining systems) in the lit and on the net.  How many
people are still using the Nordicware pressure cooker in a microwave or the
BioCare Medical electric antigen retrieval pressure cooker?  Comments are
welcome as we are looking at short and long term solutions to make the IHC
process less labor intensive.  Thanks.


Joe

From michael.ho <@t> sickkids.ca  Thu May 21 09:41:39 2009
From: michael.ho <@t> sickkids.ca (michael.ho@sickkids.ca)
Date: Fri May 22 16:39:44 2009
Subject: [Histonet] Re: [IHCRG] HRP-labeled primary antibodies
In-Reply-To: <905854.25089.qm@web50310.mail.re2.yahoo.com>
Message-ID: 

Personally, considering the low sensitivity of the direct method, I would
just treat your antibody as if it where not conjugated, and do a species
specific polymer or ABC/LSAB method. This way you can avoid Tyramide
amplification unless really necessary....Like any new antibody, you will
have to optimize your method to give you correct specificity and
sensitivity...........

regards

Michael  Ho, MLT
Resource Technologist
Immunopathology laboratory
Division of Pathology
DPLM
The Hospital for Sick Children
Toronto, Ontario
Canada
Tel#:416-813-5950
Fax: 416-813-5974


                                                                           
             Kim Merriam                                                   
                                                                 To 
             Sent by:                  Histonet                            
             ihcrg@googlegroup          
             s.com                     , ihcrg@googlegroups.com            
                                                                        cc 
                                                                           
             2009-05-21 08:41                                      Subject 
             AM                        [IHCRG] HRP-labeled primary         
                                       antibodies                          
                                                                           
             Please respond to                                             
             kmerriam2003@yaho                                             
                   o.com                                                   
                                                                           
                                                                           




Hi All,

Has anyone had experience doing IHC on FFPE tissues with HRP-labeled
primary antibodies?  I was wondering what the best way to detect them would
be.  I assume that going strait to DAB would not work, since no
amplification is there.  I was thinking of using a biotinyl tyramide step
to amplify the signal.

Also, do you think the final antibody concentration would need to be higher
than with traditional, unlabeled primaries?

Thanks,
Kim

Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA



From michael.ho <@t> sickkids.ca  Thu May 21 18:22:10 2009
From: michael.ho <@t> sickkids.ca (michael.ho@sickkids.ca)
Date: Fri May 22 16:39:46 2009
Subject: [Histonet] Re: [IHCRG] Re: CMV antibody
In-Reply-To: 
Message-ID: 

Hi Tony
If you are interested I can email more information...touch base with me
....what are you using at your institute???

best wishes


Michael  Ho, MLT
Resource Technologist
Immunopathology laboratory
Division of Pathology
DPLM
The Hospital for Sick Children
Toronto, Ontario
Canada
Tel#:416-813-5950
Fax: 416-813-5974


                                                                           
             "Tony Henwood"                                                
                                                                    To 
             Sent by:                  ,           
             ihcrg@googlegroup                        
             s.com                                                      cc 
                                        
                                       ,           
             05/21/2009 07:00                                      Subject 
             PM                        [IHCRG] Re: CMV antibody            
                                                                           
                                                                           
             Please respond to                                             
             anthonyh@chw.edu.                                             
                    au                                                     
                                                                           
                                                                           





Interestingly,

This data sheet does not seem to list the clone of the CMV monoclonal
antibody, or am I misreading it?


Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA




-----Original Message-----
From: ihcrg@googlegroups.com [mailto:ihcrg@googlegroups.com] On Behalf
Of michael.ho@sickkids.ca
Sent: Thursday, 21 May 2009 5:20 AM
To: JTAYLOR@meriter.com
Cc: 'histonet@lists.utsouthwestern.edu'; 'ihcrg@googlegroups.com'
Subject: [IHCRG] Re: CMV antibody


See attached data sheet, we use pepsin  enzyme digestion @ 37C for 30
minutes manually on FFPE

For Ventana Users Protease I for 16 minutes





Michael  Ho, MLT



Resource Technologist(See attached file: CMV-E.pdf) Immunopathology
laboratory Division of Pathology DPLM The Hospital for Sick Children
Toronto, Ontario Canada Tel#:416-813-5950
Fax: 416-813-5974




             "Taylor, Jean"

             
To
             Sent by:                  "'ihcrg@googlegroups.com'"

             ihcrg@googlegroup         ,

             s.com
"'histonet@lists.utsouthwestern.edu
                                       '"



             2009-05-20 03:06
cc
             PM


Subject
                                       [IHCRG] CMV antibody

             Please respond to

             JTAYLOR@meriter.c

                    om











Hi Everyone,

I'm wondering what antibody clone labs are using to detect
Cytomegalovirus by IHC.

Thank you!

Jean Taylor, HT(ASCP) QIHC
IHC Tech.
Meriter Health Services
Madison, WI 53715

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From angela_l_smith <@t> baxter.com  Fri May 22 09:28:14 2009
From: angela_l_smith <@t> baxter.com (angela_l_smith@baxter.com)
Date: Fri May 22 16:39:48 2009
Subject: [Histonet] Re: [IHCRG] Re: CMV antibody
In-Reply-To: 
Message-ID: 

I am leaving my position with Baxter but want to continue to receive 
emails from this group. Can someone tell me how to change the email 
address from my work address to my personal email address?
Thank you,
Angela

Angela L Smith, HT (ASCP)cm
Research Associate II/Histology
BioScience
Baxter Healthcare Corporation
25212 W. Illinois Route 120
WG2-2S Building
Round Lake, IL 60073
847-270-5530
847-270-5559 (fax)
The information transmitted is intended only for the person(s)or entity to which it is addressed and may contain confidential and/or legally privileged material. Delivery of this message to any person other than the intended recipient(s) is not intended in any way to waive privilege or confidentiality. Any review, retransmission, dissemination or other use of , or taking of any action in reliance upon, this information by entities other than the intended recipient is prohibited. If you receive this in error, please contact the sender and delete the material from any computer.

For Translation:

http://www.baxter.com/email_disclaimer
From Popp.Laurie <@t> mayo.edu  Fri May 22 15:12:58 2009
From: Popp.Laurie <@t> mayo.edu (Popp, Laurie A.)
Date: Fri May 22 16:39:50 2009
Subject: [Histonet] Cleaning embedding molds 
In-Reply-To: 
References: 
Message-ID: <1488342ADB74EC4E969C48BDCDE0BD98FB7689@msgebe23.mfad.mfroot.org>


 Happy Friday all and a long weekend too!

I am in a research area where all tissue received is fixed, processed,
and embedded as a general rule but we occasionally have blocks that
require re-embedding.  Today I was re-embedding very old blocks today
and some of my pans have a brown sludge in the bottom of them that is
probably degraded paraffin as the blocks were from 1970-1980 roughly.
We don't normally have a way to clean our pans but this is something
that is not going to just melt out and wipe up ( Very Sticky).  Does
anyone have any suggestion for cleaning?  

Thanks!

Laurie Popp, BA HT ( ASCP)
TACMA Shared Resources
Mayo Clinic Rochester



From brad <@t> yourbiomed.com  Fri May 22 15:13:34 2009
From: brad <@t> yourbiomed.com (Brad (YourBiomed))
Date: Fri May 22 16:39:52 2009
Subject: [Histonet] Leica UltraCut UCT Microtome operators manual wanted
Message-ID: <000c01c9db19$c9137250$5b3a56f0$@com>

Hello All,

 

I was hoping that someone had the operators manual for the Leica UltraCut
UCT Microtome.

Leica no longer has it on their website. I would be happy to purchase or
trade for another 

Manual.

 

Thank you for your valuable time and have a great weekend everyone.

 

Brad

 

P.S. It is not the EM model, UltraCut UCT is the only thing written on the
instrument.

From Popp.Laurie <@t> mayo.edu  Fri May 22 15:17:04 2009
From: Popp.Laurie <@t> mayo.edu (Popp, Laurie A.)
Date: Fri May 22 16:39:54 2009
Subject: [Histonet] Cleaning embedding molds 
Message-ID: <1488342ADB74EC4E969C48BDCDE0BD98FB768B@msgebe23.mfad.mfroot.org>

 Happy Friday all and a long weekend too!

I am in a research area where all tissue received is fixed, processed,
and embedded as a general rule but we occasionally have blocks that
require re-embedding.  Today I was re-embedding very old blocks today
and some of my pans have a brown sludge in the bottom of them that is
probably degraded paraffin as the blocks were from 1970-1980 roughly.
We don't normally have a way to clean our pans but this is something
that is not going to just melt out and wipe up ( Very Sticky).  Does
anyone have any suggestion for cleaning?  

Thanks!

Laurie Popp, BA HT ( ASCP)
TACMA Shared Resources
Mayo Clinic Rochester


Laurie Popp,  BA HT ASCP cm
TACMA Shared Services


From drvet_anjan <@t> hotmail.com  Sun May 24 15:48:57 2009
From: drvet_anjan <@t> hotmail.com (anjan kumar)
Date: Sun May 24 15:49:01 2009
Subject: [Histonet] antigen retrieval-pressure cooker-CD44- CD24-canine
	tissue
Message-ID: 


hello everyone,
                     I wanted to know the exact stepwise protocol for antigen retrieval using a pressure cooker. i tried using some protocol but there is no staining. Kindly tell me the detailed protocol, like when i should place the slides into the pressure cooker and from when i should count the time. 

And i am using mouse monoclonals CD44 & CD24 labvision in canine tissue, has anyone got experience with these antibodies as citrate buffer doesnt at all seems to work.
plz send me some information on this.

Regards,
Dr. Anjan Kumar.K.R

M.V.Sc Scholar

Dept. of Veterinary Pathology

Madras Veterinary College

Chennai-7

India

email: drvet_anjan@hotmail.com

Phone: +91-9940475801



_________________________________________________________________
Live Search extreme As India feels the heat of poll season, get all the info you need on the MSN News Aggregator
http://news.in.msn.com/National/indiaelections2009/aggregator/default.aspx
From amosbrooks <@t> gmail.com  Sun May 24 15:55:50 2009
From: amosbrooks <@t> gmail.com (Amos Brooks)
Date: Sun May 24 15:55:53 2009
Subject: [Histonet] CD8 or CD4 non Mouse primary
Message-ID: <582736990905241355v4595b4a4ub2bd666ac323639d@mail.gmail.com>

Hi,
   There is a new Rabbit Monoclonal out. I know Cell Marque carries it. I
haven't had a chance to evaluate it yet (copious spare time & all), but I
have had general good luck with rabbit monos. Check this site ...
http://www.cellmarque.com/07/p_detail.php?id=2030

All the best,
Amos Brooks


Message: 3
Date: Fri, 22 May 2009 10:22:14 -0700
From: "Mark Elliott" 
Subject: [Histonet] CD8 or CD4 non Mouse primary
To: 
Message-ID: <4A167CD6020000D60003F37F@mail.mrl.ubc.ca>
Content-Type: text/plain; charset=US-ASCII
Hi everyone.
We are needing to do a triple stain for IF using MHC Class II and CD4 and
CD8.  I have Mouse monoclonals to CD4 and CD8 and we have a goat anti-MHC.
We were wondering if anyone knows of a good CD4 or CD 8 that is made in
another species that we could use to avoid using two mouse antibodies. We
are doing this on human lung.
Thanks
Mark
From c.m.vanderloos <@t> amc.uva.nl  Mon May 25 01:42:05 2009
From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos)
Date: Mon May 25 01:42:20 2009
Subject: [Histonet] RE: CD8 or CD4 non Mouse primary
Message-ID: <9912e31f35c517eb.4a1a59dd@amc.uva.nl>

Hi Mark,Mouse CD4, clone 4B12 and Rabbit monoclonal CD8, clone SP16 is a good choice for your triple. Lots of success!Chris Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands Date: Fri, 22 May 2009 10:22:14 -0700
From: "Mark Elliott" 
Subject: [Histonet] CD8 or CD4 non Mouse primary
To: histonet@lists.utsouthwestern.edu

Hi everyone.

We are needing to do a triple stain for IF using MHC Class II and CD4 and CD8.  I have Mouse monoclonals to CD4 and CD8 and we have a goat anti-MHC.  We were wondering if anyone knows of a good CD4 or CD 8 that is made in another species that we could use to avoid using two mouse antibodies. We are doing this on human lung.

Thanks

Mark
From tifei <@t> foxmail.com  Mon May 25 02:53:20 2009
From: tifei <@t> foxmail.com (TF)
Date: Mon May 25 02:53:46 2009
Subject: [Histonet] Whole rat skull decalcification protocol
Message-ID: <200905251553152113230@foxmail.com>

Hi alle
i am writing to see if you can provide me any useful protocol in removing the calcium from whole rat skull fixed in PFA for frozen sections later on.

I noticed there is use of 12% formic acid mixed in 6.25% formalin;
                              or 4% HCl mixed with 8% formic acid;
or, in a gentle but slower way, 14% tetrasodium EDTA (pH 7.4-7.6)

Anyone has other good sugestions to do so?
Thanks!

2009-05-25 



TF 
From tgenade <@t> gmail.com  Mon May 25 06:23:44 2009
From: tgenade <@t> gmail.com (Tyrone Genade)
Date: Mon May 25 06:23:53 2009
Subject: [Histonet] Luxol fast blue-cresyl violet and DAB
Message-ID: 

Hello,

Anyone out there tried IHC/DAB on sections and thereafter stained with
luxol fast blue-cresyl violet? I'm looking at neurogensis in the brain
(optic tectum) of fish and want to be able to get a good idea of which
OT layers the DAB+ nuclei are lying in. Any other suggestions for a
counterstain that can be used on DAB stained sections?

Thanks
-- 
Tyrone Genade
http://tgenade.freeshell.org
email: tgenade@freeshell.org
tel: +27-84-632-1925 (c)
********************************************************************************
Romans 6:23: The gift of God is eternal life through Christ Jesus our Lord.
To find out how to receive this FREE gift visit http://www.alpha.org.

From chana.de.wolf <@t> gmail.com  Mon May 25 14:15:25 2009
From: chana.de.wolf <@t> gmail.com (Chana de Wolf)
Date: Mon May 25 14:15:30 2009
Subject: [Histonet] Accepting bids for EM work
Message-ID: <3f4b71f10905251215x780186bfye7383a8a9f110f7c@mail.gmail.com>

Hello,

We are accepting bids from EM labs for the processing and imaging of
10 rat brains. We will need 20-25 images (cortex and possibly
hippocampus) from each brain. Brains will be perfusion fixed, stored
in post-fixative solution, and shipped to the EM lab for secondary
fixation, processing, and imaging.

Please send any questions or bid proposals to me at chana.de.wolf@gmail.com.

Thank you,

Chana de Wolf
Advanced Neural Biosciences, Inc.

From pruegg <@t> ihctech.net  Mon May 25 15:42:41 2009
From: pruegg <@t> ihctech.net (pruegg@ihctech.net)
Date: Mon May 25 15:42:49 2009
Subject: [Histonet] antigen retrieval-pressure cooker-CD44- CD24-canine
	tissue
Message-ID: <20090525134241.f86bd30e73b823f57b516b5451216a98.49b9a841d6.wbe@email.secureserver.net>


   Anjan,

   I  am  more  of a fan of steamer or waterbath than pressure cooker, in
   my   my  tissue   pressure  is  s   boiling  seems to be t   it.

   are  you  sure  the  antibodies work on canine tissue?  When testing    abs, I have four protocols I run

   1. no pretreatment

   2. HIER with lph citrate buffer

   3. HIER with hph buffer, such as edta

   4. EIER, I will start with proteinase K for 5min.

   usually  with  one of these, if the ab x reacts with canine tissue you
   w   pretreatment  times   of them and I know    for  the  longest  incubatio   incubation  of primary) then I really w   species I am trying to use it on.



   Cheers,

   Patsy



   -------- Original Message --------
   Subject:   [Histonet]   antigen   r   CD24-canine
   tissue
   From: anjan kumar &   Date: Sun, May 24, 2009 1:48 pm
   To: tr   h   I  wanted  to know the exact stepwise protocol for antigen    using  a pressure cooker. i tried using some protocol but there is   staining.  Kindly  tell  me  the detailed protocol, like when i should
   plac   count the time   And  i  am  using  mouse monoclonals CD44 & CD24 labvision in can   tissue,  has  anyone  got  experience with these antibodies as citrate
   buff   plz send me some information on this.
   Regards,
   Dr. Anjan Kumar.K.R
   M.V.Sc Scholar
   Dept. of    Madras Veterinary College
   Chennai-7
   <   email: drvet_anjan@hotmail.com
   Phone: +91-9940475801   ___________________________________________________________   Live  Search extreme As India feels the heat of poll season, get a   the info you need on the MSN News Aggregator
   [1]http://news.in.msn.com/National/indiael   ections2009/aggregator/default.aspx___________________________________
   _____   Histonet mailing list
   Histonet@lists.utsouthwestern.edu[2]http://lists.utsouthwestern.edu/ma
   ilman/listinfo/histonet
   <
References

   1. 3D"http://news.i=/
   2. 3D"http://lists.utsouthwestern.edu/mailman/listinfo/histonet"
From CBraaten <@t> Cheshire-Med.COM  Tue May 26 04:26:25 2009
From: CBraaten <@t> Cheshire-Med.COM (Christine I. Braaten)
Date: Tue May 26 04:26:36 2009
Subject: [Histonet] cloudy formalin in vip processor
Message-ID: 

Dennis,

We do a warm water flush every Tuesday. We generally have between 40 and
120 blocks a day. Most of our specimens are GI biopsies but we do have
things like uterus, breast tissue, tonsils, things like that. Knock on
wood we have not had a blockage in the 9 years we have been using our
processor. Our biggest problem is the pathologists dissatisfaction with
how our GI biopsies look (dry, cracked). That could be because we run
them on the overnight schedule with our big surgical specimens. I
mentioned washing the specimens to one pathologist and she's not really
up for doing that. So, we'll keep trying and see what happens. Have a
great weekend. 


CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients 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 electronic mail and destroy all copies of the original message.
From Kemlo.Rogerson <@t> waht.swest.nhs.uk  Tue May 26 06:16:04 2009
From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson)
Date: Tue May 26 06:16:13 2009
Subject: [Histonet] cloudy formalin in vip tissue processor
Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06C0D3C8@wahtntex2.waht.swest.nhs.uk>

That's a 'harsh' fixative; used it years and years ago as it mordants
the haematoxylin. I think that I still stand by the harsh effects of
that fixative on coagulating protein and that the culprit is precisely
that; coagulated protein from using a additive fixative (as you know
formalin is a coagulant, non-additive fixative, normally).



Kemlo Rogerson	
e-mail kemlorogerson@nhs.net if not at work.			
DD   01934 647057 or extension 3311     Mob 07749 754194; 
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer
This e-mail is confidential and privileged. If you are not the intended
recipient please accept my apologies; please do not disclose, copy or
distribute information in this e-mail or take any action in reliance on
its contents: to do so is strictly prohibited and may be unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation

 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Christine I. Braaten
Sent: 22 May 2009 17:36
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] cloudy formalin in vip tissue processor

Thanks to all who have replied to my query. I probably should have
mentioned that we use zinc formalin in our processor and as our primary
fixative. I have spoken to both Sakura and Anatech (supplier of zinc
formalin) and feel more confused. Anatech says it's a change in pH that
causes the zinc to precipitate and that xylene will cause a whole lot
more problems than a little cloudiness. Sakura thinks the zinc formalin
is the problem. We clean the retort with the cleaning program every
morning and make sure to dry the retort so I'm not sure how xylene would
get into the formalin. If this brings up any other issues I would
appreciate more feedback. Thanks 


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contain confidential and privileged information.  Any unauthorized
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the intended recipient, please contact the sender by electronic mail and
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From Kemlo.Rogerson <@t> waht.swest.nhs.uk  Tue May 26 06:19:00 2009
From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson)
Date: Tue May 26 06:19:05 2009
Subject: [Histonet] Bone!
Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06C0D3CC@wahtntex2.waht.swest.nhs.uk>

As I say I don't have experience of preservatives but question if the
effects of ethanol (70%) over a long time may also 'overfix' the bone?
Anyway 70% ethanol has perfectly preserved me!!


Kemlo Rogerson	
e-mail kemlorogerson@nhs.net if not at work.			
DD   01934 647057 or extension 3311     Mob 07749 754194; 
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer
This e-mail is confidential and privileged. If you are not the intended
recipient please accept my apologies; please do not disclose, copy or
distribute information in this e-mail or take any action in reliance on
its contents: to do so is strictly prohibited and may be unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation

 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rittman,
Barry R
Sent: 22 May 2009 13:21
To: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Bone!

Lets face it there is no really ideal medium for long term storage.
However this depends on what techniques you want to carry out later and
when.
I agree that 10% NBF is a good fixative for bone, I would definitely not
advise leaving it in this fixative for extended periods of time.
The reason is that formalin continues cross linking.
This means that many stains and immunohistochemistry may no longer be
possible depending on the amount of time left in fixative.
Also Kemlo is correct in that temporary formalin bonds can be broken by
extended washing in watre. This only applies to the weak temporary bonds
and not to the more stable bonds. The longer the fixing the greater the
number of stable bonds.
I would recommend having a mixture of 70% ethanol plus glycerin up to
10-20%.  The glycerin will prevent any drying out of the solution over
extended periods of tim.
Barry


________________________________________
From: histonet-bounces@lists.utsouthwestern.edu
[histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kemlo Rogerson
[Kemlo.Rogerson@waht.swest.nhs.uk]
Sent: Friday, May 22, 2009 2:24 AM
To: Robert Edward Pogue
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Bone!

Probably of all the fixatives 10% neutral buffered formalin is least
likely to overfix and I've left tissues in it for years. Formalin, as
you know, is one of those fixatives that you can wash out. There are
preservatives, rather than fixatives, you could try but I don't have
experience of those; my best advise is formalin but make sure that it is
neutral and buffered. I think the effects of pH would be more damaging
than the relatively 'soft' formalin fixation.



Kemlo Rogerson
e-mail kemlorogerson@nhs.net if not at work.
DD   01934 647057 or extension 3311     Mob 07749 754194;
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer This e-mail is confidential and privileged. If you are not the
intended recipient please accept my apologies; please do not disclose,
copy or distribute information in this e-mail or take any action in
reliance on its contents: to do so is strictly prohibited and may be
unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation



-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert
Edward Pogue
Sent: 21 May 2009 16:33
To: Kemlo Rogerson
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Bone!

Thanks Kemlo...

I thought of that, but was worried about over-fixation,- what do you
think?

Redward.


------------------------------

In 10% neutral buffered formalin?



Kemlo Rogerson
e-mail kemlorogerson@nhs.net if not at work.
DD   01934 647057 or extension 3311     Mob 07749 754194;
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer This e-mail is confidential and privileged. If you are not the
intended recipient please accept my apologies; please do not disclose,
copy or distribute information in this e-mail or take any action in
reliance on its contents: to do so is strictly prohibited and may be
unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation



-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert
Edward Pogue
Sent: 21 May 2009 14:47
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Bone!

Friends,

Can someone recommend to me how to store bones (juvenile rat femur) that
I have fixed and decalcified, but am not yet ready to cut (they're for
vibratome, so not embedded).

Thanks!

Redward.

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From aj.taylor <@t> blueyonder.co.uk  Tue May 26 06:52:06 2009
From: aj.taylor <@t> blueyonder.co.uk (alan taylor)
Date: Tue May 26 06:52:10 2009
Subject: [Histonet] Slide Clips for Shandon Linistain GLX
Message-ID: <5B9982E86DAF4900A5D512B88D1A4D02@merlin>

Hi to everyone in Histoland

We have a Shandon Linistain GLX that we use for our H & E staining. The problem we have is that the plastic clips used to attach the slides to the chain drive are no longer available. The stubs have broken on many of our clips preventing their attachment to the chain links and rendering them unusable. We depend heavily on this machine for our work and now have only a small supply of useable clips.

Does anyone know of a possible 'second supplier' source of these clips, anywhere in the world? Or does anyone have any of these clips gathering dust in a store room somewhere that we could use. Any assistance or information would be greatly appreciated.   

Alan Taylor BSc(Hons), FRMS
Microtechnical Services
Exeter. Devon. UK
From rjr6 <@t> psu.edu  Tue May 26 07:38:03 2009
From: rjr6 <@t> psu.edu (Roberta Horner)
Date: Tue May 26 07:38:20 2009
Subject: [Histonet] RE: Cleaning embedding molds 
In-Reply-To: <1488342ADB74EC4E969C48BDCDE0BD98FB768B@msgebe23.mfad.mfroot.org>
References: <1488342ADB74EC4E969C48BDCDE0BD98FB768B@msgebe23.mfad.mfroot.org>
Message-ID: 

To clean my molds I periodically put them in water with soap and bring it to a boil.  Then I set the pan somewhere to cool and once the paraffin hardens I take it off then rinse the molds with a lot of water, dry and spray with mold release.  I don't need to do this too often but I like doing it in the winter so I can set the hot pan outside to cool
Roberta Horner HTL/HT
Penn State University
Animal Diagnostic Lab

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Popp, Laurie A.
Sent: Friday, May 22, 2009 4:17 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Cleaning embedding molds 

 Happy Friday all and a long weekend too!

I am in a research area where all tissue received is fixed, processed,
and embedded as a general rule but we occasionally have blocks that
require re-embedding.  Today I was re-embedding very old blocks today
and some of my pans have a brown sludge in the bottom of them that is
probably degraded paraffin as the blocks were from 1970-1980 roughly.
We don't normally have a way to clean our pans but this is something
that is not going to just melt out and wipe up ( Very Sticky).  Does
anyone have any suggestion for cleaning?  

Thanks!

Laurie Popp, BA HT ( ASCP)
TACMA Shared Resources
Mayo Clinic Rochester


Laurie Popp,  BA HT ASCP cm
TACMA Shared Services


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From Kemlo.Rogerson <@t> waht.swest.nhs.uk  Tue May 26 08:11:25 2009
From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson)
Date: Tue May 26 08:11:31 2009
Subject: [Histonet] cloudy formalin in vip tissue processor
Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06C0D471@wahtntex2.waht.swest.nhs.uk>

hehehe, thanks...
 
Only got it completely the wrong way around didn't I? Freida is correct,
I think!
 

Kemlo Rogerson  

e-mail kemlorogerson@nhs.net   if not at
work.                    

DD   01934 647057 or extension 3311     Mob 07749 754194; 

Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
Lehrer

This e-mail is confidential and privileged. If you are not the intended
recipient please accept my apologies; please do not disclose, copy or
distribute information in this e-mail or take any action in reliance on
its contents: to do so is strictly prohibited and may be unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation


 

 


________________________________

From: Freida Carson [mailto:freidac@sbcglobal.net] 
Sent: 26 May 2009 13:38
To: Kemlo Rogerson
Subject: RE: [Histonet] cloudy formalin in vip tissue processor


I'm sorry, but formalin is an additive, non-coagulant fixative.  I am
not sending this through the histonet, but did want to correct you.  I
imagine others will also.  Formalin adds on at the amino group.
 
Freida Carson

--- On Tue, 5/26/09, Kemlo Rogerson 
wrote:


	
	From: Kemlo Rogerson 
	Subject: RE: [Histonet] cloudy formalin in vip tissue processor
	To: "Christine I. Braaten" ,
histonet@lists.utsouthwestern.edu
	Date: Tuesday, May 26, 2009, 6:16 AM
	
	
	That's a 'harsh' fixative; used it years and years ago as it
mordants
	the haematoxylin. I think that I still stand by the harsh
effects of
	that fixative on coagulating protein and that the culprit is
precisely
	that; coagulated protein from using a additive fixative (as you
know
	formalin is a coagulant, non-additive fixative, normally).
	
	
	
	Kemlo Rogerson    
	e-mail kemlorogerson@nhs.net
  if
not at work.            
	DD   01934 647057 or extension 3311     Mob 07749 754194; 
	Embrace uncertainty. Hard problems rarely have easy solutions.
--Jonah
	Lehrer
	This e-mail is confidential and privileged. If you are not the
intended
	recipient please accept my apologies; please do not disclose,
copy or
	distribute information in this e-mail or take any action in
reliance on
	its contents: to do so is strictly prohibited and may be
unlawful.
	Please inform me that this message has gone astray before
deleting it.
	Thank you for your co-operation
	
	
	
	-----Original Message-----
	From: histonet-bounces@lists.utsouthwestern.edu
 
	[mailto:histonet-bounces@lists.utsouthwestern.edu
 ] On Behalf Of
	Christine I. Braaten
	Sent: 22 May 2009 17:36
	To: histonet@lists.utsouthwestern.edu
 
	Subject: [Histonet] cloudy formalin in vip tissue processor
	
	Thanks to all who have replied to my query. I probably should
have
	mentioned that we use zinc formalin in our processor and as our
primary
	fixative. I have spoken to both Sakura and Anatech (supplier of
zinc
	formalin) and feel more confused. Anatech says it's a change in
pH that
	causes the zinc to precipitate and that xylene will cause a
whole lot
	more problems than a little cloudiness. Sakura thinks the zinc
formalin
	is the problem. We clean the retort with the cleaning program
every
	morning and make sure to dry the retort so I'm not sure how
xylene would
	get into the formalin. If this brings up any other issues I
would
	appreciate more feedback. Thanks 
	
	
	CONFIDENTIALITY NOTICE: This electronic message, including any
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	review, use, disclosure or distribution is prohibited.  If you
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From fudo <@t> ufl.edu  Tue May 26 08:40:04 2009
From: fudo <@t> ufl.edu (FU,DONGTAO)
Date: Tue May 26 08:40:12 2009
Subject: [Histonet] autoflurescence of cytospin cells
Message-ID: <2123048600.400051243345204465.JavaMail.osg@osgjas04.cns.ufl.edu>

Hi, all

  I need you help.I was doing immunoflurescent staining on 
cytospin mouse tumor cells and I noticed high autoflurescence 
background of the cells, it happened even before I added first 
antibody. I tried 4% paraformaldehyde or acetone fixation, but 
either of them gave me the same result.

  Does anyone here have experiences on decreasing the 
autoflurescence on the cytospin cells? Or any suggestions? The PI 
wants to see double staining, so it is impossible to do chromagen 
methods.

  Thank you,


Ann Dongtao Fu
University of Flodrida


From JBower <@t> hei.org  Tue May 26 10:26:48 2009
From: JBower <@t> hei.org (Bower, Jennifer)
Date: Tue May 26 10:26:55 2009
Subject: [Histonet] Laser tweezer
Message-ID: <87449E4A2B01DA47B29424CE5D6E0F830C56CA6C@hi0sml1.hei.org>

   Does anyone have an instruction manual for a Cell Robotics Laser
microdissection/ Laser Tweezer system? I would love to get a copy of the
instructions. Thanks for you help.

 

Jennifer Bower, HTL (ASCP)

Histology Core, CB&G

House Ear Institute

2100 W 3rd Street, 5th Floor

Los Angeles, CA 90057

(213) 989-7460

 

From Herrick.James <@t> mayo.edu  Tue May 26 11:08:30 2009
From: Herrick.James <@t> mayo.edu (Herrick, James L. (Jim))
Date: Tue May 26 11:08:36 2009
Subject: [Histonet] Question on Sanderson's Rapid Bone Stain
Message-ID: <7267A64D75F58241B577876D8A885631191F09@msgebe41>

Hello everyone,

I am trying to run an SRBS on a 5 ?m thick, deplasticized (MMA) section of rabbit calvaria. I have had really good luck with thick sections (plastic left on), but cannot get a good stain on the thin, deplasticized section. Does anyone have any protocols they don't mind sharing? Your help is and has always been greatly appreciated. Thanks again.

Jim
From sjchtascp <@t> yahoo.com  Tue May 26 11:34:22 2009
From: sjchtascp <@t> yahoo.com (Steven Coakley)
Date: Tue May 26 11:34:25 2009
Subject: [Histonet] Looking for an HT position
Message-ID: <411189.80250.qm@web38201.mail.mud.yahoo.com>

So.WI/No.Ill.? Will work FT, PT, FTE or just as needed to cover staffing shortages.
Thanks everyone,
?
Steve Coakley
Beloit, WI


      
From lpaveli1 <@t> hurleymc.com  Tue May 26 13:16:18 2009
From: lpaveli1 <@t> hurleymc.com (Lynette Pavelich)
Date: Tue May 26 13:16:40 2009
Subject: [Histonet] IHC control debate
Message-ID: <4A1BF9B2020000EE0002970A@smtp-gw.hurleymc.com>

Hello,
I need help from all of our IHC expert guru's!  We have a debate going
on about the known positive control blocks.  All are FFPE with 8-48 hrs
in formalin.  All have been tested and have been approved for use as
positive controls.  We currently cut the positive control from the block
as needed.
I have learned that some controls do start to deteriorate over time,
tonsils for example.  But, is there a list, an article, SOMETHING that
can guide me so that we can cut some of these controls ahead of time? 
This would not only save tech time, but would further the use of our
controls.  In cutting the block, some of the good control is wasted and
some positives are hard to find.
My argument is that the purchased controls work wonderfully, even after
a very long time and our improved detection kits have improved so much
over the years, it makes this possible.

If there is no published article out there, I implore you experts to
please help us in this area.  Would also like to hear from the technical
experts from our suppliers too!  Until then, the debate continues
on!!!!!!!

Thank you!!
Lynette

From LSebree <@t> uwhealth.org  Tue May 26 13:40:58 2009
From: LSebree <@t> uwhealth.org (Sebree Linda A)
Date: Tue May 26 13:41:03 2009
Subject: [Histonet] IHC control debate
In-Reply-To: <4A1BF9B2020000EE0002970A@smtp-gw.hurleymc.com>
Message-ID: <5998F3BDFF7AAC4091C7AE93A7A1A5892CE00C@UWHC-MAIL01.uwhis.hosp.wisc.edu>

Lynette,

Its not so much the positive control tissue as the antigen it's
detecting; there are antigens that are quite labile and so lose their
reactivity over time when the positive control tissue slides are stored
at room temperature.  Off the top of my head, ER, PR, Ki67, bcl-2, bcl-6
are some that come to mind...there are a number of others.  So we store
some previously cut control slides at RT and others at -20 degrees C
depending upon the antigen they will be used for.  You can find further
information regarding the more sensitive antigens in the Histonet
Archives.

Hope this helps,

Linda A. Sebree
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
DB1-223 VAH
600 Highland Ave.
Madison, WI 53792
(608)265-6596



-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lynette
Pavelich
Sent: Tuesday, May 26, 2009 1:16 PM
To: histonet@lists.utsouthwestern.edu; fudo@ufl.edu
Subject: [Histonet] IHC control debate


Hello,
I need help from all of our IHC expert guru's!  We have a debate going
on about the known positive control blocks.  All are FFPE with 8-48 hrs
in formalin.  All have been tested and have been approved for use as
positive controls.  We currently cut the positive control from the block
as needed. I have learned that some controls do start to deteriorate
over time, tonsils for example.  But, is there a list, an article,
SOMETHING that can guide me so that we can cut some of these controls
ahead of time? 
This would not only save tech time, but would further the use of our
controls.  In cutting the block, some of the good control is wasted and
some positives are hard to find. My argument is that the purchased
controls work wonderfully, even after a very long time and our improved
detection kits have improved so much over the years, it makes this
possible.

If there is no published article out there, I implore you experts to
please help us in this area.  Would also like to hear from the technical
experts from our suppliers too!  Until then, the debate continues
on!!!!!!!

Thank you!!
Lynette

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

From SDrew <@t> uwhealth.org  Tue May 26 13:55:43 2009
From: SDrew <@t> uwhealth.org (Drew Sally A)
Date: Tue May 26 13:55:48 2009
Subject: [Histonet] IHC control debate
In-Reply-To: <4A1BF9B2020000EE0002970A@smtp-gw.hurleymc.com>
Message-ID: <738A7878143FF74BB77436E255743C1A1F7BCF@UWHC-MAIL03.uwhis.hosp.wisc.edu>

I think that besides checking Histonet archives for past discussions on
this, if you keep a file of freshly cut, freshly stained control slides
that you can go back to compare day-to-day results with, you will be
able to pinpoint fading stains better.

You may also want to have cut control slides inventory that coincides
with an estimated workload-for example, have 1 months-worth of tonsil at
room-temp and 2-3 months in a frost-free freezer.  If you have some at
both storage conditions, then you can check the staining quality versus
the control section's age as you optimize antibodies.

 

Although this is a generalization-most of the stains we've seen affected
by control age/storage conditions have been nuclear-staining antibodies.

 

Sally Ann Drew, MT (ASCP)

sdrew@uwhealth.org

 

IHC/ISH Lab DB1-223, Mail Code 3224

600 Highland Ave.

Madison, WI 53792

Phone (608) 265-6596

Fax (608) 262-7174

 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lynette
Pavelich
Sent: Tuesday, May 26, 2009 1:16 PM
To: histonet@lists.utsouthwestern.edu; fudo@ufl.edu
Subject: [Histonet] IHC control debate

 

Hello,

I need help from all of our IHC expert guru's!  We have a debate going

on about the known positive control blocks.  All are FFPE with 8-48 hrs

in formalin.  All have been tested and have been approved for use as

positive controls.  We currently cut the positive control from the block

as needed.

I have learned that some controls do start to deteriorate over time,

tonsils for example.  But, is there a list, an article, SOMETHING that

can guide me so that we can cut some of these controls ahead of time? 

This would not only save tech time, but would further the use of our

controls.  In cutting the block, some of the good control is wasted and

some positives are hard to find.

My argument is that the purchased controls work wonderfully, even after

a very long time and our improved detection kits have improved so much

over the years, it makes this possible.

 

If there is no published article out there, I implore you experts to

please help us in this area.  Would also like to hear from the technical

experts from our suppliers too!  Until then, the debate continues

on!!!!!!!

 

Thank you!!

Lynette

 

_______________________________________________

Histonet mailing list

Histonet@lists.utsouthwestern.edu

http://lists.utsouthwestern.edu/mailman/listinfo/histonet

From CIngles <@t> uwhealth.org  Tue May 26 18:03:48 2009
From: CIngles <@t> uwhealth.org (Ingles Claire )
Date: Tue May 26 18:03:52 2009
Subject: [Histonet] Slide Clips for Shandon Linistain GLX
References: <5B9982E86DAF4900A5D512B88D1A4D02@merlin>
Message-ID: 

Alan:
I hope your first supplier isn't Shandon. I do have a part number through them, but don't know how much they are. If anyone else knows of a second supplier, let me know too. I'm in the same boat as Alan. SOMEBODY should still sell these, as they still sell the stainers.
The part number I have is 6754002. This number should be good as of a couple years ago. These are listed as the while clips, though I think there are green and pink ones too. We don't really care what color they are, just so they work! Don't even get me started on trying to get a replacement chain drive motor... (I think I have that number too if anyone needs it.)
 
Claire

________________________________

From: histonet-bounces@lists.utsouthwestern.edu on behalf of alan taylor
Sent: Tue 5/26/2009 6:52 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Slide Clips for Shandon Linistain GLX



Hi to everyone in Histoland

We have a Shandon Linistain GLX that we use for our H & E staining. The problem we have is that the plastic clips used to attach the slides to the chain drive are no longer available. The stubs have broken on many of our clips preventing their attachment to the chain links and rendering them unusable. We depend heavily on this machine for our work and now have only a small supply of useable clips.

Does anyone know of a possible 'second supplier' source of these clips, anywhere in the world? Or does anyone have any of these clips gathering dust in a store room somewhere that we could use. Any assistance or information would be greatly appreciated.  

Alan Taylor BSc(Hons), FRMS
Microtechnical Services
Exeter. Devon. UK
_______________________________________________
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Histonet@lists.utsouthwestern.edu
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From AnthonyH <@t> chw.edu.au  Tue May 26 18:44:35 2009
From: AnthonyH <@t> chw.edu.au (Tony Henwood)
Date: Tue May 26 18:44:47 2009
Subject: [Histonet] cloudy formalin in vip processor
In-Reply-To: 
Message-ID: 

Chris,

Often the cracking is due to incomplete formalin fixation combined with
too harsh processing. We found a marked improvement when we extended the
fixation time and decreased the time in the processing solutions.

Having a rotary processor also helps. GI and other small biopsie are
processed on the rotary processor whilst the larger, thicker specimen
are processed in the retort processor.

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Christine I. Braaten
Sent: Tuesday, 26 May 2009 7:26 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] cloudy formalin in vip processor


Dennis,

We do a warm water flush every Tuesday. We generally have between 40 and
120 blocks a day. Most of our specimens are GI biopsies but we do have
things like uterus, breast tissue, tonsils, things like that. Knock on
wood we have not had a blockage in the 9 years we have been using our
processor. Our biggest problem is the pathologists dissatisfaction with
how our GI biopsies look (dry, cracked). That could be because we run
them on the overnight schedule with our big surgical specimens. I
mentioned washing the specimens to one pathologist and she's not really
up for doing that. So, we'll keep trying and see what happens. Have a
great weekend. 


CONFIDENTIALITY NOTICE: This electronic message, including any
attachments, is for the sole use of the intended recipients 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 electronic mail and
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_______________________________________________
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This note also confirms that this email message has been
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From AnthonyH <@t> chw.edu.au  Tue May 26 18:57:47 2009
From: AnthonyH <@t> chw.edu.au (Tony Henwood)
Date: Tue May 26 18:57:53 2009
Subject: [Histonet] IHC control debate
In-Reply-To: <4A1BF9B2020000EE0002970A@smtp-gw.hurleymc.com>
Message-ID: 

Here is one reference that might be of use:

Philippe Bertheau "Variability of immunohistochemical reactivity on
stored paraffin slides" J Clin Pathol 1998;51:370-374

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lynette
Pavelich
Sent: Wednesday, 27 May 2009 4:16 AM
To: histonet@lists.utsouthwestern.edu; fudo@ufl.edu
Subject: [Histonet] IHC control debate


Hello,
I need help from all of our IHC expert guru's!  We have a debate going
on about the known positive control blocks.  All are FFPE with 8-48 hrs
in formalin.  All have been tested and have been approved for use as
positive controls.  We currently cut the positive control from the block
as needed. I have learned that some controls do start to deteriorate
over time, tonsils for example.  But, is there a list, an article,
SOMETHING that can guide me so that we can cut some of these controls
ahead of time? 
This would not only save tech time, but would further the use of our
controls.  In cutting the block, some of the good control is wasted and
some positives are hard to find. My argument is that the purchased
controls work wonderfully, even after a very long time and our improved
detection kits have improved so much over the years, it makes this
possible.

If there is no published article out there, I implore you experts to
please help us in this area.  Would also like to hear from the technical
experts from our suppliers too!  Until then, the debate continues
on!!!!!!!

Thank you!!
Lynette

_______________________________________________
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 note also confirms that this email message has been
virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses.
**********************************************************************


From hlukey <@t> msn.com  Tue May 26 20:48:18 2009
From: hlukey <@t> msn.com (Hugh Luk)
Date: Tue May 26 20:48:24 2009
Subject: [Histonet] Slide Clips for Shandon Linistain GLX
In-Reply-To: 
References: <5B9982E86DAF4900A5D512B88D1A4D02@merlin>
	
Message-ID: 


Alan,

I use these too, but I'm not sure what is available.  I found the web address at:

www1.fishersci.com/wps/portal/PRODUCTDETAIL?productId=9097251&catalogId=29103&pos=25&catCode=HC_SC&fromCat=yes&keepSessionSearchOutPut=true&brCategoryId=57660&hlpi=y&fromSearch=
 
is the Fisher Linear stainer (as Fisher bought out ThermoShandon).  Click on accessories.  It appears to list pink, blue and white stainer clips are available for $87.12 (for 25).  I don't know why white is expensive or why green and yellow are not available.

I've been looking at other plastic manufacturers (Simport, Marketlab, ect.), but I have not found another vendor.  I hope this helps and I wish you (us) luck.

Hugh
KP-Hawaii/
Cancer research center of Hawaii

> Date: Tue, 26 May 2009 18:03:48 -0500
> From: CIngles@uwhealth.org
> To: Histonet@lists.utsouthwestern.edu
> Subject: RE: [Histonet] Slide Clips for Shandon Linistain GLX
> CC: 
> 
> Alan:
> I hope your first supplier isn't Shandon. I do have a part number through them, but don't know how much they are. If anyone else knows of a second supplier, let me know too. I'm in the same boat as Alan. SOMEBODY should still sell these, as they still sell the stainers.
> The part number I have is 6754002. This number should be good as of a couple years ago. These are listed as the while clips, though I think there are green and pink ones too. We don't really care what color they are, just so they work! Don't even get me started on trying to get a replacement chain drive motor... (I think I have that number too if anyone needs it.)
>  
> Claire
> 
> ________________________________
> 
> From: histonet-bounces@lists.utsouthwestern.edu on behalf of alan taylor
> Sent: Tue 5/26/2009 6:52 AM
> To: Histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Slide Clips for Shandon Linistain GLX
> 
> 
> 
> Hi to everyone in Histoland
> 
> We have a Shandon Linistain GLX that we use for our H & E staining. The problem we have is that the plastic clips used to attach the slides to the chain drive are no longer available. The stubs have broken on many of our clips preventing their attachment to the chain links and rendering them unusable. We depend heavily on this machine for our work and now have only a small supply of useable clips.
> 
> Does anyone know of a possible 'second supplier' source of these clips, anywhere in the world? Or does anyone have any of these clips gathering dust in a store room somewhere that we could use. Any assistance or information would be greatly appreciated.  
> 
> Alan Taylor BSc(Hons), FRMS
> Microtechnical Services
> Exeter. Devon. UK
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> 
> 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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From Maxim_71 <@t> mail.ru  Tue May 26 21:00:33 2009
From: Maxim_71 <@t> mail.ru (Maxim Peshkov)
Date: Tue May 26 21:01:02 2009
Subject: [Histonet] Modifications of human and viral DNA by formaldehyde
	fixation
Message-ID: <1324408654.20090527060033@mail.ru>

Dear colleagues!
One friend of mine can not able to find full text of the article:
Karlsen F, Kalantari M, Chitemerere M, et al.
Modifications of human and viral deoxyribonucleic
acid by formaldehyde fixation. Lab Invest 1994;71:604?11.
Can anyone help to us?
Many thanks.

Maxim Peshkov,
Russia,
Taganrog.
                          mailto:Maxim_71@mail.ru


From gu.lang <@t> gmx.at  Wed May 27 04:32:49 2009
From: gu.lang <@t> gmx.at (Gudrun Lang)
Date: Wed May 27 04:32:59 2009
Subject: AW: [Histonet] IHC control debate
In-Reply-To: <4A1BF9B2020000EE0002970A@smtp-gw.hurleymc.com>
References: <4A1BF9B2020000EE0002970A@smtp-gw.hurleymc.com>
Message-ID: <9B11ACA232114CAEB4739FE96E1A382E@dielangs.at>

Here is another publication:
Preservation of Estrogen Receptor in Paraffin Sections, Christine M. Bromley
et al; J. Histotech. 17:115, 1994

Gudrun Lang

-----Urspr?ngliche Nachricht-----
Von: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Lynette
Pavelich
Gesendet: Dienstag, 26. Mai 2009 20:16
An: histonet@lists.utsouthwestern.edu; fudo@ufl.edu
Betreff: [Histonet] IHC control debate

Hello,
I need help from all of our IHC expert guru's!  We have a debate going
on about the known positive control blocks.  All are FFPE with 8-48 hrs
in formalin.  All have been tested and have been approved for use as
positive controls.  We currently cut the positive control from the block
as needed.
I have learned that some controls do start to deteriorate over time,
tonsils for example.  But, is there a list, an article, SOMETHING that
can guide me so that we can cut some of these controls ahead of time? 
This would not only save tech time, but would further the use of our
controls.  In cutting the block, some of the good control is wasted and
some positives are hard to find.
My argument is that the purchased controls work wonderfully, even after
a very long time and our improved detection kits have improved so much
over the years, it makes this possible.

If there is no published article out there, I implore you experts to
please help us in this area.  Would also like to hear from the technical
experts from our suppliers too!  Until then, the debate continues
on!!!!!!!

Thank you!!
Lynette

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


From tgenade <@t> gmail.com  Wed May 27 07:44:55 2009
From: tgenade <@t> gmail.com (Tyrone Genade)
Date: Wed May 27 07:45:04 2009
Subject: [Histonet] LFB and DAB: do they play nice together?
Message-ID: 

Hello,

Sorry to be a pest, but normally responses come quite quick to queries
so I'm figuring that maybe my last query never registered (this being
the wake of Memorial Day weekend and all...).

Has anyone out there tried DAB on sections and thereafter stained with
luxol fast blue-cresyl violet? I'm looking at neurogensis in the brain
of fish and want to be able to get a good idea of which layers the
DAB+ nuclei are lying in. Any other suggestions for a counterstain
that can be used on DAB stained sections?

I'm adjusting the DAB color with Cobalt Chloride, which should change
it from brown to blue-black---maybe not the best idea with LFB. What
about when using copper or nickle?

Thanks
-- 
Tyrone Genade
http://tgenade.freeshell.org
email: tgenade@freeshell.org
tel: +27-84-632-1925 (c)
********************************************************************************
Romans 6:23: The gift of God is eternal life through Christ Jesus our Lord.
To find out how to receive this FREE gift visit http://www.alpha.org.

From DixonM <@t> vetmed.ufl.edu  Wed May 27 10:24:15 2009
From: DixonM <@t> vetmed.ufl.edu (MaryAnn Dixon)
Date: Wed May 27 10:24:23 2009
Subject: [Histonet] test
Message-ID: <530D827EC657DE418C3572ADD63FCDC3249811@EXGVMCNETWORK.vetmed.ufl.edu>

This is a test as I am not receiving email from histonet.

MaryAnn Dixon BS, HT(ASCP)
Biological Scientist
Oncology
UF Veterinary Medical Center
(352) 392-2226 Ext. 5739

From Debra.Ortiz <@t> uchospitals.edu  Wed May 27 11:10:36 2009
From: Debra.Ortiz <@t> uchospitals.edu (Debra.Ortiz@uchospitals.edu)
Date: Wed May 27 11:10:46 2009
Subject: [Histonet] CJD procedures
Message-ID: <5392DB699B157E4C8E65B3779634BD7D011AF58B@uchmbx04-hpk03s.UCHAD.uchospitals.edu>

We are looking to revamp our procedure for CJD and I would like to see
if there are different protocols out there. At this point we are
handling the whole process by hand, including melting the paraffin in a
melting pot. Do we need a separate oven for melting the paraffin and
heating the slides?

Thank you

 

 

Debra Ann Ortiz

Chief Medical Technologist

The University of Chicago Medical Center

Room E-602-A

5841 S. Maryland Avenue

Chicago, Il 60637

phone: 773.702.5237

 

********************************************************************************
This e-mail is intended only for the use of the individual or entity to which
it is addressed and may contain information that is privileged and confidential.
If the reader of this e-mail message is not the intended recipient, you are 
hereby notified that any dissemination, distribution or copying of this
communication is prohibited. If you have received this e-mail in error, please 
notify the sender and destroy all copies of the transmittal. 

Thank you
University of Chicago Medical Center 
********************************************************************************
From mari.ann.mailhiot <@t> leica-microsystems.com  Wed May 27 11:34:18 2009
From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com)
Date: Wed May 27 11:34:27 2009
Subject: [Histonet] CJD procedures
In-Reply-To: <5392DB699B157E4C8E65B3779634BD7D011AF58B@uchmbx04-hpk03s.UCHAD.uchospitals.edu>
Message-ID: 

Debra

Contact Case Western University. They have National Pathology Survelience
Prion Disease site - www.cjdsurveillance.com. This is a excellent resource

Best Regards

Mari Ann Mailhiot BA HT ASCP
Application Specialist/Trainer
Leica Microsystems
Biosystems Division
Technical Assistance Center
800 248 0123 x7267
847 236 3063 fax
mari.ann.mailhiot@leica-microsystems.com
www.leica-microsystems.com


                                                                           
                                                               
             Sent by:                                                   To 
             histonet-bounces@          
             lists.utsouthwest                                          cc 
             ern.edu                                                       
                                                                   Subject 
                                       [Histonet] CJD procedures           
             05/27/2009 11:10                                              
             AM                                                            
                                                                           
                                                                           
                                                                           
                                                                           



We are looking to revamp our procedure for CJD and I would like to see
if there are different protocols out there. At this point we are
handling the whole process by hand, including melting the paraffin in a
melting pot. Do we need a separate oven for melting the paraffin and
heating the slides?

Thank you





Debra Ann Ortiz

Chief Medical Technologist

The University of Chicago Medical Center

Room E-602-A

5841 S. Maryland Avenue

Chicago, Il 60637

phone: 773.702.5237



********************************************************************************

This e-mail is intended only for the use of the individual or entity to
which
it is addressed and may contain information that is privileged and
confidential.
If the reader of this e-mail message is not the intended recipient, you are
hereby notified that any dissemination, distribution or copying of this
communication is prohibited. If you have received this e-mail in error,
please
notify the sender and destroy all copies of the transmittal.

Thank you
University of Chicago Medical Center
********************************************************************************

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Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


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From Michael.Owen <@t> fda.hhs.gov  Wed May 27 11:44:45 2009
From: Michael.Owen <@t> fda.hhs.gov (Owen, Michael P)
Date: Wed May 27 11:44:52 2009
Subject: [Histonet] CJD procedures
In-Reply-To: <5392DB699B157E4C8E65B3779634BD7D011AF58B@uchmbx04-hpk03s.UCHAD.uchospitals.edu>
Message-ID: <449E51C6DA0AD840B44F57C7A6EB07BF09267EF8@FMD3VS022.fda.gov>

The American Biological Safety Association (ABSA) and its Biosafety
Mailing List are two excellent resources for obtaining information on
handling CJD materials.

American Biological Safety Association (ABSA)
http://www.absa.org

ABSA - Resources and Tools - E-Mail Groups
http://www.absa.org/restool.html
http://www.absa.org/resgroups.html


ABSA is also useful for any other question about biological safety. Its
members have practical knowledge of laboratory operations. :o)




Michael P. Owen, Regulatory Microbiologist
U.S. FDA Pacific Regional Lab Northwest
22201 23rd Drive SE  Bothell, WA 98021-4421
Phone: 425-483-4865     E-Mail: michael.owen@fda.hhs.gov

From Sarah.Yates <@t> STJUDE.ORG  Tue May 26 10:50:45 2009
From: Sarah.Yates <@t> STJUDE.ORG (Yates, Sarah)
Date: Wed May 27 12:17:12 2009
Subject: [Histonet] Tennessee Society for Histotechnology State Meeting
Message-ID: 

The TnSH would like to announce the 2009 meeting held in Memphis. We have two full days of workshops covering safety, alternative embedding methods, Immunohistochemistry, xylene free processing, basic photography for anatomic pathology, special stains, bone preparation, research histology techniques, diagnostic panels and laboratory management. Please join us to meet your continuing education requirements while making new friends and experiencing the Memphis atmosphere down on Beale Street with our famous BBQ and Blues music!
For more information on workshops and registration, go to www.tnsh.org and click on the 2009 TSH Meeting Program Brochure link. You can print the registration pages and mail to:

Rhonda Schalk
TSH Treasurer
1675 Pleasant Road
Evensville, TN 37332

OR

Charlene Henry
c/o St. Jude Children's Research Hospital
262 Danny Thomas Place M/S 250
Memphis, Tn 38105

________________________________
Email Disclaimer: www.stjude.org/emaildisclaimer
From hborgeri <@t> wfubmc.edu  Wed May 27 12:44:40 2009
From: hborgeri <@t> wfubmc.edu (Hermina Borgerink)
Date: Wed May 27 12:45:56 2009
Subject: [Histonet] Custom ISH probes
Message-ID: <9AEEF1FB6254224AA355ED285F849165377802E7@EXCHVS2.medctr.ad.wfubmc.edu>

We are interested in having a custom ISH probe prepared using a specific sequence for which we already have the necessary information. Anyone know of a reliable company who does this?  Vendors are welcome to contact me.

Hermina

Hermina M. Borgerink, BA, HT, HTL(ASCP)QIHC
Wake Forest University Primate Center
Department of Pathology
Medical Center Blvd.
Winston-Salem, NC 27157
Tel. (336) 716-1538
Fax. (336) 716-1515
e-mail:? hborgeri@wfubmc.edu



From jtaylor <@t> meriter.com  Wed May 27 12:52:15 2009
From: jtaylor <@t> meriter.com (Taylor, Jean)
Date: Wed May 27 12:52:19 2009
Subject: [Histonet] CD21
Message-ID: <466B666475DE6547BBB0641E540A4BB504C3F5C476@EXVS1.meriter.com>

Hi everyone,

We are looking into ordering the CD21 antibody and was wondering what antibody clone labs are using.

Thanks!

Jean Taylor, HT(ASCP)QIHC
IHC Tech
Meriter Health Services
Madison, WI 53715
From rhbrown1 <@t> histocs.com  Wed May 27 11:23:45 2009
From: rhbrown1 <@t> histocs.com (Leroy Brown)
Date: Wed May 27 13:07:34 2009
Subject: [Histonet] help needed to find parts for tissue processor
Message-ID: 

Hi ,  I have two MVP-1 tissue processor  that are still in great shape except that both have lost the display  screens.   Other than that they are wonderful tissue  processors.    I am told by the folks that "know" this  machine that I can swap out these screens from any working model from  any of the following processors.   The RMC models,  the MVP-1  or MVP-2 the older 1530 , the Renaissance processor.   They are  all similar since this machine changed hands so many times.  It was a  Fisher Scientific,  then Instrumentation Lab,  then Vantana  and so on.   The problem is that the parts are not available any  longer.    My only hope if I want to continue using these  processors is to find a parts machine or one that is sitting in the basement  of someones hospital or lab perhaps for years.  So, if you have one or  know where to find one I would very much like to hear from you.    I bought these two processors about 2 years ago from a  government auction still new in the ori
ginal crate  unopened.    I really would like to get them  up and running again.   

Thanks  

LeRoy Brown HT(ASCP) HTL  

www.histocs.com  

email to me directly at lhbhcs@comcast.net or call me at  360-966-7300
From mmcgraw <@t> phenopath.com  Wed May 27 13:35:48 2009
From: mmcgraw <@t> phenopath.com (Medea McGraw)
Date: Wed May 27 13:36:00 2009
Subject: [Histonet] Melanin / melanocytes staining method tintorial or IHC
Message-ID: 

Craig,

I was searching the archives on the histonet concerning the Dopa Oxidase
stain. I have not performed this stain but have done the Schmorls stain. I
was wondering if you had a particular method/protocol that you use for this
stain? Are there any kits available or is it cheaper to do buying the
reagents separately? Any suggestions would be greatly appreciated.

Regards,
Medea


-- 
Medea J. McGraw BS, HT, HTL(ASCP) CM
IHC Research Histotechnologist
Phenopath Laboratories
Phone: 206-374-9000 x1033
Fax: 206-374-9009



This e-mail message, including any attachments, is for the sole use of the 
intended recipients and may contain privileged information. Any unauthorized 
review, use, disclosure or distribution is prohibited. If you are not the intended 
recipient, please contact the sender by e-mail and destroy all copies of the 
original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. 
at (206) 374-9000.
From histo20 <@t> hotmail.com  Wed May 27 14:14:03 2009
From: histo20 <@t> hotmail.com (Paula Wilder)
Date: Wed May 27 14:14:08 2009
Subject: [Histonet] Full Time Histology Position Opening in Towson, Md
Message-ID: 


Hello Histonet,

 

St. Joseph Medical Center in Towson, Md. is looking for a full time Histology Technician for day shift.  If interested, please contact Paula Wilder, Histology Supervisor at 410-337-1741.  

_________________________________________________________________
Hotmail? has ever-growing storage! Don?t worry about storage limits.
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From AnthonyH <@t> chw.edu.au  Wed May 27 19:10:13 2009
From: AnthonyH <@t> chw.edu.au (Tony Henwood)
Date: Wed May 27 19:10:26 2009
Subject: [Histonet] RE: [IHCRG] CD21
In-Reply-To: <466B666475DE6547BBB0641E540A4BB504C3F5C476@EXVS1.meriter.com>
Message-ID: 

Jean,
 
Here is the details from our manual
 

ANTIGEN NAME:


CD 21


 

OTHER NAMES:        

 

Clone:             2G9

Isotype:          IgG2a

 

DESCRIPTION:         Monoclonal antibody to a Prokaryotic recombinant
protein corresponding to the external domain of the CD21 molecule.

          

STORAGE CONDITIONS:        4-8oC undiluted

 

 

SUPPLIER:                Novocastra NCL-CD21-2G9 

 


PROCEDURE:


 

Methodology:        HRP-POLYMER

Working Dilution:    1/20        Bond   1/40

Special Conditions:      HIER (pH6 - Citrate)

Frozen Section: 

CONTROL TISSUE:   Lymph Node/Tonsil               

 

Inbuilt Controls                       Lymphocytes

 

 

 


CLINICAL SIGNIFICANCE:


CD21 antigen is a type I integral membrane glycoprotein of molecular
weight 140kD. The CD21 molecule, present on mature B cells, is involved
in transmitting growth-promoting signals to the interior of the B cell
and acts as a receptor for Epstein-Barr virus. The antigen is absent on
T-lymphocytes, monocytes and granulocytes.


REFERENCES:


 

Ling N R, Brown B and Hardie D (1994) Journal of Immunological Methods.
173: 11-17 

Timens W, Boes A, Vos H, et al (1991) Histochemistry. 95: 605-611 

Regards 

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 


	-----Original Message-----
	From: ihcrg@googlegroups.com [mailto:ihcrg@googlegroups.com] On
Behalf Of Taylor, Jean
	Sent: Thursday, 28 May 2009 3:52 AM
	To: 'ihcrg@googlegroups.com';
'histonet@lists.utsouthwestern.edu'
	Subject: [IHCRG] CD21
	
	

	Hi everyone,

	 

	We are looking into ordering the CD21 antibody and was wondering
what antibody clone labs are using.

	 

	Thanks!

	 

	Jean Taylor, HT(ASCP)QIHC

	IHC Tech

	Meriter Health Services

	Madison, WI 53715


*********************************************************************
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 note 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 gu.lang <@t> gmx.at  Thu May 28 02:12:36 2009
From: gu.lang <@t> gmx.at (Gudrun Lang)
Date: Thu May 28 02:12:51 2009
Subject: AW: [Histonet] Melanin / melanocytes staining method tintorial or IHC
In-Reply-To: 
References: 
Message-ID: <62FBE1C6607A4C9F9B5020BABF7EF2F7@dielangs.at>


There's a silverstain for melanin and other argentaffine granules:
Fontana-Masson.
The principle is based on the reducing effect of melanin on
silverdiaminions.
You can find the procedure in the AFIP manual.
Gudrun

-----Urspr?ngliche Nachricht-----
Von: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Medea
McGraw
Gesendet: Mittwoch, 27. Mai 2009 20:36
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Melanin / melanocytes staining method tintorial or IHC

Craig,

I was searching the archives on the histonet concerning the Dopa Oxidase
stain. I have not performed this stain but have done the Schmorls stain. I
was wondering if you had a particular method/protocol that you use for this
stain? Are there any kits available or is it cheaper to do buying the
reagents separately? Any suggestions would be greatly appreciated.

Regards,
Medea


-- 
Medea J. McGraw BS, HT, HTL(ASCP) CM
IHC Research Histotechnologist
Phenopath Laboratories
Phone: 206-374-9000 x1033
Fax: 206-374-9009



This e-mail message, including any attachments, is for the sole use of the 
intended recipients and may contain privileged information. Any unauthorized

review, use, disclosure or distribution is prohibited. If you are not the
intended 
recipient, please contact the sender by e-mail and destroy all copies of the

original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A.

at (206) 374-9000.
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


From ree3 <@t> leicester.ac.uk  Thu May 28 03:28:32 2009
From: ree3 <@t> leicester.ac.uk (Edwards, R.E.)
Date: Thu May 28 03:28:40 2009
Subject: [Histonet] muscle striations
Message-ID: <7722595275A4DD4FA225B92CDBF174A18C50DBF4E0@EXC-MBX3.cfs.le.ac.uk>


Any favourite methods for the above??  I'm aware of Heidenhain's and Mallory's PTAH.
                           Many thanks
                              Richard Edwards
                                    University of Leicester

From d.a.faichney <@t> stir.ac.uk  Thu May 28 04:56:10 2009
From: d.a.faichney <@t> stir.ac.uk (Deborah Faichney)
Date: Thu May 28 04:56:23 2009
Subject: [Histonet] Horseshoe crab..help!
Message-ID: <8ED3F2CA5B78E142B8193376C57330F8E198E18F55@EXCH2007.ad.stir.ac.uk>

Hi all,

I have a Masters student here who has Horseshoe crab tissues fixed in 10% NBF.  Most of the tissues have been sectioned successfully but the eyes are surrounded by armour plated chitin! (she broke two Dremel drills trying to cut the carapace and finally tin snips had to be used to cut them out!!).
Any thoughts on how to soften them?  Looking around on the Net, I have found  that processing and clearing through Chloroform may soften, also a solution called Diaphanol has been recommended.
At the moment they have been two days in a 1 part acetic acid to 5 parts NBF with no effect.
We only have a couple of weeks to do this work so cannot adopt any trial or lengthy methods.

Is this possible?  I like a challenge, but suspect this needs a miracle.  Its no wonder these beasties have been on the planet for so long.
Many thanks

Debbie Faichney
Histopathology
Institute of Aquaculture
University of Stirling
Stirling, FK7 7QS
Scotland
Uk




-- 
Academic Excellence at the Heart of Scotland.
The University of Stirling is a charity registered in Scotland, 
 number SC 011159.

From TMcNemar <@t> lmhealth.org  Thu May 28 05:12:57 2009
From: TMcNemar <@t> lmhealth.org (Tom McNemar)
Date: Thu May 28 05:14:28 2009
Subject: [Histonet] Certification CEUs
Message-ID: <51D5D78FBEDAEA4FBCCD9A9D44211DC505B8E0EE@lmhsmail.lmhealth.org>

Hello all,

I'm sure this has been discussed but I can't seem to find it.....

What are people doing to meet the requirements for CEUs?  Is there a list of acceptable programs or methods of aquiring the required CEUs?  I never had to worry about it but I now have a newly certified tech who will need them.  Thanks in advance for any input.

Tom McNemar, HT(ASCP)
Histology Co-ordinator
Licking Memorial Health Systems
(740) 348-4163
(740) 348-4166
tmcnemar@lmhealth.org
www.LMHealth.org


From lpwenk <@t> sbcglobal.net  Thu May 28 06:04:40 2009
From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk)
Date: Thu May 28 06:04:53 2009
Subject: [Histonet] Certification CEUs
In-Reply-To: <51D5D78FBEDAEA4FBCCD9A9D44211DC505B8E0EE@lmhsmail.lmhealth.org>
Message-ID: 

ASCP has a booklet for the Certification Maintenance Program (CMP) that
lists types of CE, how many hours each, and through what agencies. 
http://ascp.org/pdf/CMPBooklet.aspx
If that doesn't work, go to
www.ascp.org  -->  Laboratory Professional --> CMP --> #1 Booklet. Look at
pages 6-8.

It does list NSH. But certificates from state society meetings can also be
used. In-services at your lab can also be used, just have to document -
title, presenter, date, amount of time, name of institution, have it signed
by someone in charge. Sign in sheets with this information could be used, if
you don't want to make certificates for each in-service. Teleconferences
count, as do sending in the tests from the articles in magazines/journals
such as MLO, Advance or Journal of Histotechnology.

Just a little plug for NSH Teleconferences - $125 for each session, and you
can have as many people attend and get CEU for each. About 2 months after
the teleconference, you get a CD of the handouts, PowerPoint and speaker,
with a test for the teleconference you signed up for. You can use this up to
2 years later, and people can still earn CEU (for those people on vacation,
had to stay back in the lab to work, you hired 6 months later.) 

NSH Teleconferences are 4th Wednesday of the month, 1-2 pm Eastern time, for
1 hour-ish. The next presentations are Troubleshooting IHC, Staining of
Pigments and Minerals, and Role of IHC in Lymphomas.
http://www.nsh.org/TC/2009TCbrochure.pdf
If that doesn't work, go to
www.nsh.org --> Meetings/Events --> NSH Events --> 2009 Teleconference

Disclaimer - I am the NSH Teleconference coordinator. No, I don't get a
kickback from getting more labs to sign up for the NSH Teleconferences. Just
the satisfaction of knowing that histotechs around the US, Canada, and
sometimes other countries, are getting continuing education. 

Peggy A. Wenk, HTL(ASCP)SLS
Beaumont Hospital
Royal Oak, MI 48073

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tom McNemar
Sent: Thursday, May 28, 2009 6:13 AM
To: histonet@pathology.swmed.edu
Subject: [Histonet] Certification CEUs

Hello all,

I'm sure this has been discussed but I can't seem to find it.....

What are people doing to meet the requirements for CEUs?  Is there a list of
acceptable programs or methods of aquiring the required CEUs?  I never had
to worry about it but I now have a newly certified tech who will need them.
Thanks in advance for any input.

Tom McNemar, HT(ASCP)
Histology Co-ordinator
Licking Memorial Health Systems
(740) 348-4163
(740) 348-4166
tmcnemar@lmhealth.org
www.LMHealth.org


_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


From bhewlett <@t> cogeco.ca  Thu May 28 06:59:25 2009
From: bhewlett <@t> cogeco.ca (Bryan Hewlett)
Date: Thu May 28 06:59:31 2009
Subject: [Histonet] muscle striations
References: <7722595275A4DD4FA225B92CDBF174A18C50DBF4E0@EXC-MBX3.cfs.le.ac.uk>
Message-ID: 

Richard,

My quick and easy test for muscle striations is to check the H&E under 
crossed polarizers.
Myosin is birefringent. The birefringence can be intensified by staining 
with picric acid, e.g. following a Van Gieson stain.

Regards,

Bryan

----- Original Message ----- 
From: "Edwards, R.E." 
To: "Histonet" 
Sent: Thursday, May 28, 2009 4:28 AM
Subject: [Histonet] muscle striations



Any favourite methods for the above??  I'm aware of Heidenhain's and 
Mallory's PTAH.
                           Many thanks
                              Richard Edwards
                                    University of Leicester

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



From igor.deyneko <@t> gmail.com  Thu May 28 10:04:18 2009
From: igor.deyneko <@t> gmail.com (Igor Deyneko)
Date: Thu May 28 10:04:24 2009
Subject: [Histonet] Alpha Smooth Muscle Actin in Xenografts
Message-ID: <35e16a770905280804n15a325d8k5c627feb755a0a33@mail.gmail.com>

Dear Histonetters!
I was wondering if anyone out there is working with xenograft tumor models
and knows of a good alpha smooth muscle actin antibody that will detect
mouse tissue but would give minimal background in tumor cells??? I have
tried 6 different ones, 3 from Biocare, 1 from Santa Cruz, and 2 from Abcam,
but they all gave me very strong backgrounds. Any information would be
greatly appreciated.
Thank you.
Igor Deyneko
Infinity Pharmaceuticals
Cambridge, MA 02139
From akemiat3377 <@t> yahoo.com  Thu May 28 11:14:25 2009
From: akemiat3377 <@t> yahoo.com (akemiat3377@yahoo.com)
Date: Thu May 28 11:14:35 2009
Subject: [Histonet] Recycling Formalin for Fresh Tissue
Message-ID: <715329.45426.qm@web31306.mail.mud.yahoo.com>

Good Morning Histo Land,

I am asking all you out there to give me your input on recycIing formalin.? I realize this has been discussed in the not too distant past, but this may be a little different situation.? I realize some labs are recycling formalin to put on their tissue processors, but this is a totally different scenario.

I have been a manager at my lab for only a month.? Much to my surprise, I just found out yesterday that our lab is recycling formalin and shipping it out to one of our clients to put their FRESH SPECIMENS into.? 

I just had a lengthy conversation with Robert Lott the day before yesterday, regarding pre-analytical protocol's effecting the test results of AFB.? By the way, we were in full agreement.? This formalin issue was unknown to me then.? I realize that the pre-analytical, pre-analytical, pre-analytical, process effects the final out come.? I need to present my case to our Director of Pathology why this is compromising patient care.

Any and all responses are gladly welcome.

Thanks,
Akemi

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287 
W: E-Mail: aallison-tacha@apmglab.com
P: E-Mail: akemiat3377@yahoo.com

From pattennj <@t> mail.nih.gov  Thu May 28 11:49:24 2009
From: pattennj <@t> mail.nih.gov (Patten, Nicole (NIH/NIAAA) [F])
Date: Thu May 28 11:49:29 2009
Subject: [Histonet] Hypoxia in Post-Mortem Samples
Message-ID: 

Hey there!

I have some post-mortem brain samples (FFPE) with varying PMI's and I'd like to determine the hypoxia levels (and location) at time of death (to identify if this may play a role in the disease I am studying). First of all, does anyone have any suggestions as to a good hypoxia antibody? Second of all, does anyone know how the PMI would affect the staining for hypoxia?

Thanks, I'd appreciate any comments!

Nicole J. Patten
Post-Baccalaureate Fellow/IRTA
National Institutes of Health
From Carrie <@t> nsh.org  Thu May 28 12:34:05 2009
From: Carrie <@t> nsh.org (Carrie Diamond)
Date: Thu May 28 12:34:12 2009
Subject: [Histonet] RE: Certification CEUs
References: 
Message-ID: 

Hello Tom,

The National Society of Histotechnology has a number of different
options for maintaining certification. Membership in the society enables
you a possible 6 contact hours per year for participating in reading
select articles and quizzes. These are included with your membership and
will get you halfway to your re-certification in 3 years. 

https://www.nshonline.org/eweb/DynamicPage.aspx?expires=yes&Site=NSH&Web
Key=bb7290f9-de36-4e1e-b6a7-c83609ce6d38


Additionally, we offer a Teleconference Series which may be beneficial
to your entire lab. 11 possible hours are available per year. Please see
included link.
http://www.nsh.org/organizations.php3?action=printContentTypeHome&orgid=
111&typeID=1380


We also offer a number of events that provide the face to face education
that is so valuable.
http://www.nsh.org/organizations.php3?action=printContentTypeHome&orgid=
111&typeID=1380


If you or your colleagues are unable to attend an event, we have a
number of courses archived in our Live Learning Center, which are
available online and the attendee can be a little more flexible with
their participation. 
http://www.softconference.com/NSH/default.asp


Please feel free to contact me directly with any questions about the
above mentioned programs. Good luck in your pursuit of continuing
education. 

Thank you, 

Carrie Diamond
Executive Director
National Society for Histotechnology
10320 Little Patuxent Parkway
Suite 804
Columbia, MD 21044
P: 443.535.4060
Direct: 443.535.4066
Fax: 443.535.4055
E-mail: carrie@nsh.org


Save the Date!
35th Annual Symposium Convention
October 3-8, 2009
Birmingham, AL
Registration Details available in March
www.nsh.org 



------------------------------

Message: 11
Date: Thu, 28 May 2009 06:12:57 -0400
From: "Tom McNemar" 
Subject: [Histonet] Certification CEUs
To: 
Message-ID:
	<51D5D78FBEDAEA4FBCCD9A9D44211DC505B8E0EE@lmhsmail.lmhealth.org>
Content-Type: text/plain;	charset="iso-8859-1"

Hello all,

I'm sure this has been discussed but I can't seem to find it.....

What are people doing to meet the requirements for CEUs?  Is there a
list of acceptable programs or methods of aquiring the required CEUs?  I
never had to worry about it but I now have a newly certified tech who
will need them.  Thanks in advance for any input.

Tom McNemar, HT(ASCP)
Histology Co-ordinator
Licking Memorial Health Systems
(740) 348-4163
(740) 348-4166
tmcnemar@lmhealth.org
www.LMHealth.org




From gu.lang <@t> gmx.at  Thu May 28 12:34:45 2009
From: gu.lang <@t> gmx.at (Gudrun Lang)
Date: Thu May 28 12:34:52 2009
Subject: AW: [Histonet] Recycling Formalin for Fresh Tissue
In-Reply-To: <715329.45426.qm@web31306.mail.mud.yahoo.com>
References: <715329.45426.qm@web31306.mail.mud.yahoo.com>
Message-ID: <2D92C3B7D45940128A8CBFBFFD55B857@dielangs.at>

Just for interest. Your lab takes the recycled formalin to make 4% neutral
buffered formalin (NBF)? Or do they ship the recycled formalin directly in
bottles to the surgery?
AFB = acid fast bacili?

Do you have concerns, that they produce false-positive AFB stains? I cannot
imagine, that a (hopefully) small number of dead bacili on the surface of
tissue will pretend this. An other question: Isn't there a filter in the
recycler to prevent such contamination?
Gudrun

-----Urspr?ngliche Nachricht-----
Von: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von
akemiat3377@yahoo.com
Gesendet: Donnerstag, 28. Mai 2009 18:14
An: Histonet
Betreff: [Histonet] Recycling Formalin for Fresh Tissue

Good Morning Histo Land,

I am asking all you out there to give me your input on recycIing formalin.?
I realize this has been discussed in the not too distant past, but this may
be a little different situation.? I realize some labs are recycling formalin
to put on their tissue processors, but this is a totally different scenario.

I have been a manager at my lab for only a month.? Much to my surprise, I
just found out yesterday that our lab is recycling formalin and shipping it
out to one of our clients to put their FRESH SPECIMENS into.? 

I just had a lengthy conversation with Robert Lott the day before yesterday,
regarding pre-analytical protocol's effecting the test results of AFB.? By
the way, we were in full agreement.? This formalin issue was unknown to me
then.? I realize that the pre-analytical, pre-analytical, pre-analytical,
process effects the final out come.? I need to present my case to our
Director of Pathology why this is compromising patient care.

Any and all responses are gladly welcome.

Thanks,
Akemi

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287 
W: E-Mail: aallison-tacha@apmglab.com
P: E-Mail: akemiat3377@yahoo.com

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


From Vickroy.Jim <@t> mhsil.com  Thu May 28 12:42:47 2009
From: Vickroy.Jim <@t> mhsil.com (Vickroy, Jim)
Date: Thu May 28 12:42:53 2009
Subject: [Histonet] GUIDELINES FOR HER2NEU FIXATION
Message-ID: <24A4826E8EF0964D86BC5317306F58A52BBB293EE7@mmc-mail.ad.mhsil.com>


Last year I understood that the guidelines for formalin fixation of breast tissue for Her2neu recommended 6 - 48 hours.  I believe the guidelines left some discretion with needle biopsies also.  I am now hearing that the guidelines are going to or are already changed or restated.  Does anybody have any new information?

Jim




Jim Vickroy BS, HT(ASCP)
Technical Supervisor - Surgical and Autopsy Pathology
Memorial Medical Center
217-788-4046
vickroy.jim@mhsil.com



This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited.

From akemiat3377 <@t> yahoo.com  Thu May 28 13:21:25 2009
From: akemiat3377 <@t> yahoo.com (Akemi Allison-Tacha)
Date: Thu May 28 13:21:30 2009
Subject: Fw: Re: AW: [Histonet] AFB & Recycling Formalin for Fresh Tissue 
Message-ID: <966137.39305.qm@web31306.mail.mud.yahoo.com>

Hello,
I responded to Gudrun personally, but I thought that the information I am supplying him below might be a good educational source for other histologists who are having similar scenario's for AFB contamination.? 

I would also like to say that I have an extremely eager histology team that have expressed an interest in learning theory and practice.? Most of my team are OJT.? They have had several pathologists approach them with these concerns regarding AFB, and because of their knowledge, or lack of it, couldn't fix the problem.? We are starting with the basics and I am giving in-services along the way.

Hi  Gudrun,

They are shipping the recycled formalin directly to the outside hospital surgery to put their fresh tissues into.....

As far as the AFB, we have had intermitant (+) AFB on the edges of tissues, that has been extremely questionable.? We currently are not cutting a negative (-) & positive(+) control on the same water bath as the test sample. That will be remedied beginning Monday. The water here is also in question.? I will be sending it out for microbacteria analysis.

We are going through a thorough housekeeping and in-service process.? We are starting with the tissue processors, which are currently only changed weekly (not rotated daily), embedding centers, which have to my knowledge, never been drained, and the hopper cleaned-out.? The water baths are not scrubbed out with soap and HOT water daily and
 covered at night to prevent contamination.? The water bath is not skimmed of excess tissue debris, each and every time another specimen is placed on it.? The Carbol Fuchsin is not being filtered prior to use.? The working solution is currently being poured back into the stock bottle.? Need I go on................

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287 
W: E-Mail: aallison-tacha@apmglab.com
P: E-Mail: akemiat3377@yahoo.com



--- On Thu, 5/28/09, Gudrun Lang  wrote:

From: Gudrun Lang 
Subject: AW: [Histonet] Recycling Formalin for Fresh Tissue
To: akemiat3377@yahoo.com
Cc:
 histonet@lists.utsouthwestern.edu
Date: Thursday, May 28, 2009, 10:34 AM

Just for interest. Your lab takes the recycled formalin to make 4% neutral
buffered formalin (NBF)? Or do they ship the recycled formalin directly in
bottles to the surgery?
AFB = acid fast bacili?

Do you have concerns, that they produce false-positive AFB stains? I cannot
imagine, that a (hopefully) small number of dead bacili on the surface of
tissue will pretend this. An other question: Isn't there a filter in the
recycler to prevent such contamination?
Gudrun

-----Urspr?ngliche Nachricht-----
Von: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von
akemiat3377@yahoo.com
Gesendet: Donnerstag, 28. Mai 2009 18:14
An: Histonet
Betreff: [Histonet] Recycling Formalin for Fresh Tissue

Good Morning Histo Land,

I am asking all you out there to give me your input on recycIing formalin.?
I realize this has been discussed in the not too distant past, but this may
be a little different situation.? I realize some labs are recycling formalin
to put on their tissue processors, but this is a totally different scenario.

I have been a manager at my lab for only a month.? Much to my surprise, I
just found out yesterday that our lab is recycling formalin and shipping it
out to one of our clients to put their FRESH SPECIMENS into.?
 

I just had a lengthy conversation with Robert Lott the day before yesterday,
regarding pre-analytical protocol's effecting the test results of AFB.? By
the way, we were in full agreement.? This formalin issue was unknown to me
then.? I realize that the pre-analytical, pre-analytical, pre-analytical,
process effects the final out come.? I need to present my case to our
Director of Pathology why this is compromising patient care.

Any and all responses are gladly welcome.

Thanks,
Akemi

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287 
W: E-Mail: aallison-tacha@apmglab.com
P: E-Mail: akemiat3377@yahoo.com

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

From freidac <@t> sbcglobal.net  Thu May 28 14:09:18 2009
From: freidac <@t> sbcglobal.net (Freida Carson)
Date: Thu May 28 14:09:24 2009
Subject: Fw: Re: AW: [Histonet] AFB & Recycling Formalin for Fresh Tissue
Message-ID: <335684.74616.qm@web82507.mail.mud.yahoo.com>

Wang reported on the contamination of tissue sections with AFB by the use of fluorescence microscopy, and?if I remember correctly, when he tested the water in water fountains, he found something like 33% contained AFB.? Non-pathogenic, but we can't tell that on our stains. The paper is in Am J Clin Pathol 51:71, 1969.? We (Carson, Kingsley, Haberman, and Race) also reported the contamination in the 1964 issue of the same journal.? We stopped using tap water for our water baths and began using millipore filtered water that had already been through a deionizing column and charcoal filter.? We always cut a negative control from the same days workload as the patient case - uterus in our case.? You don't need a positive control cut the same day, just a positive control with only a medium?number of organisms. ?We also?did not use any tap water in the deparaffinization prior to the carbol fuchsin.? AFB organisms have also been reported growing in 40
 gal formalin tanks believe it or not and in the old?Fisher?Paraffin wafers.
?
We centrifuged?tap water in 50-ml tubes, poured off the supernatent, and refilled and recentrifuged until we could see some sediment in the bottom.? We took that to Microbioloby and had it cultured to definited prove the presence of AFB in the tap water.
I?would recommend this approach to anyone suspecting?tissue contamination.?
?
Freida Carson
--- On Thu, 5/28/09, Akemi Allison-Tacha  wrote:


From: Akemi Allison-Tacha 
Subject: Fw: Re: AW: [Histonet] AFB & Recycling Formalin for Fresh Tissue
To: "Histonet" 
Date: Thursday, May 28, 2009, 1:21 PM


Hello,
I responded to Gudrun personally, but I thought that the information I am supplying him below might be a good educational source for other histologists who are having similar scenario's for AFB contamination.? 

I would also like to say that I have an extremely eager histology team that have expressed an interest in learning theory and practice.? Most of my team are OJT.? They have had several pathologists approach them with these concerns regarding AFB, and because of their knowledge, or lack of it, couldn't fix the problem.? We are starting with the basics and I am giving in-services along the way.

Hi? Gudrun,

They are shipping the recycled formalin directly to the outside hospital surgery to put their fresh tissues into.....

As far as the AFB, we have had intermitant (+) AFB on the edges of tissues, that has been extremely questionable.? We currently are not cutting a negative (-) & positive(+) control on the same water bath as the test sample. That will be remedied beginning Monday. The water here is also in question.? I will be sending it out for microbacteria analysis.

We are going through a thorough housekeeping and in-service process.? We are starting with the tissue processors, which are currently only changed weekly (not rotated daily), embedding centers, which have to my knowledge, never been drained, and the hopper cleaned-out.? The water baths are not scrubbed out with soap and HOT water daily and
covered at night to prevent contamination.? The water bath is not skimmed of excess tissue debris, each and every time another specimen is placed on it.? The Carbol Fuchsin is not being filtered prior to use.? The working solution is currently being poured back into the stock bottle.? Need I go on................

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287 
W: E-Mail: aallison-tacha@apmglab.com
P: E-Mail: akemiat3377@yahoo.com



--- On Thu, 5/28/09, Gudrun Lang  wrote:

From: Gudrun Lang 
Subject: AW: [Histonet] Recycling Formalin for Fresh Tissue
To: akemiat3377@yahoo.com
Cc:
histonet@lists.utsouthwestern.edu
Date: Thursday, May 28, 2009, 10:34 AM

Just for interest. Your lab takes the recycled formalin to make 4% neutral
buffered formalin (NBF)? Or do they ship the recycled formalin directly in
bottles to the surgery?
AFB = acid fast bacili?

Do you have concerns, that they produce false-positive AFB stains? I cannot
imagine, that a (hopefully) small number of dead bacili on the surface of
tissue will pretend this. An other question: Isn't there a filter in the
recycler to prevent such contamination?
Gudrun

-----Urspr?ngliche Nachricht-----
Von: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von
akemiat3377@yahoo.com
Gesendet: Donnerstag, 28. Mai 2009 18:14
An: Histonet
Betreff: [Histonet] Recycling Formalin for Fresh Tissue

Good Morning Histo Land,

I am asking all you out there to give me your input on recycIing formalin.?
I realize this has been discussed in the not too distant past, but this may
be a little different situation.? I realize some labs are recycling formalin
to put on their tissue processors, but this is a totally different scenario.

I have been a manager at my lab for only a month.? Much to my surprise, I
just found out yesterday that our lab is recycling formalin and shipping it
out to one of our clients to put their FRESH SPECIMENS into.?


I just had a lengthy conversation with Robert Lott the day before yesterday,
regarding pre-analytical protocol's effecting the test results of AFB.? By
the way, we were in full agreement.? This formalin issue was unknown to me
then.? I realize that the pre-analytical, pre-analytical, pre-analytical,
process effects the final out come.? I need to present my case to our
Director of Pathology why this is compromising patient care.

Any and all responses are gladly welcome.

Thanks,
Akemi

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287 
W: E-Mail: aallison-tacha@apmglab.com
P: E-Mail: akemiat3377@yahoo.com

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
From sweaver <@t> tvmdl.tamu.edu  Thu May 28 14:17:34 2009
From: sweaver <@t> tvmdl.tamu.edu (Stephanie Weaver)
Date: Thu May 28 14:18:13 2009
Subject: [Histonet] Re: horseshoe crabs
In-Reply-To: <20090528171631.C6D7A1C3B2@sr-4-int.cis.tamu.edu>
References: <20090528171631.C6D7A1C3B2@sr-4-int.cis.tamu.edu>
Message-ID: <4A1E9CFE.A3DC.00A0.0@tvmdl.tamu.edu>

I have no experience with horseshoe crabs, but we do process many aquatic animal specimens.  We have used Davidson's fixative (acetic-alcohol-formalin) to soften the exoskeleton.  Post-fixation after 10% NBF works as well, since many of our clients do not have anything but formalin on hand.  I'm not sure how well it would work on something as large and solid as a horseshoe crab, but we have had success with fish, shrimp, and hermit crabs.

Stephanie Weaver
Texas Veterinary Medical Diagnostic Laboratory

Message: 10
Date: Thu, 28 May 2009 10:56:10 +0100
From: Deborah Faichney 
Subject: [Histonet] Horseshoe crab..help!
To: "histonet@lists.utsouthwestern.edu"
	
Message-ID:
	<8ED3F2CA5B78E142B8193376C57330F8E198E18F55@EXCH2007.ad.stir.ac.uk>
Content-Type: text/plain; charset="us-ascii"

Hi all,

I have a Masters student here who has Horseshoe crab tissues fixed in 10% NBF.  Most of the tissues have been sectioned successfully but the eyes are surrounded by armour plated chitin! (she broke two Dremel drills trying to cut the carapace and finally tin snips had to be used to cut them out!!).
Any thoughts on how to soften them?  Looking around on the Net, I have found  that processing and clearing through Chloroform may soften, also a solution called Diaphanol has been recommended.
At the moment they have been two days in a 1 part acetic acid to 5 parts NBF with no effect.
We only have a couple of weeks to do this work so cannot adopt any trial or lengthy methods.

Is this possible?  I like a challenge, but suspect this needs a miracle.  Its no wonder these beasties have been on the planet for so long.
Many thanks

Debbie Faichney
Histopathology
Institute of Aquaculture
University of Stirling
Stirling, FK7 7QS
Scotland
Uk







From david.kinsley <@t> spcorp.com  Thu May 28 14:20:51 2009
From: david.kinsley <@t> spcorp.com (Kinsley, David)
Date: Thu May 28 14:20:55 2009
Subject: [Histonet] Advice on fatty liver sectioning
Message-ID: 

Hi,
I have some very fatty mouse livers that I have to obtain frozen sections for Oil red O and IHC.  I have tried sectioning from -18 to -30 degrees and from 5um to 10um and the liver just seems to compress, pull away from the OCT, or roll up on itself while the OCT sections well.  When I section below -30 parts of the liver have chatter in them as if they are too cold.  Does anyone have any suggestions on what the best cutting temperature would be for these?

I have successfully cut normal and less fatty livers at 5um ad 10 um at -18 and -25 degrees respectively, it is these extremely fatty livers that are giving me problems.  Any advice is appreciated.

Dave


 


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From carmen_loiselle <@t> hotmail.com  Thu May 28 14:22:30 2009
From: carmen_loiselle <@t> hotmail.com (carmen loiselle)
Date: Thu May 28 14:22:35 2009
Subject: [Histonet] Multispecimen tissue blocks /sausage
Message-ID: 


To all histonetters,

 

I need your expertise for the preparation of sausage multispecimen block for quality control in immunoshistochemistry lab.  At first my pathologist wanted to try microarray but because of the cost he had decided to go for sausage instead.  I would greatly appreciate any information regarding this matter starting with the procurement of the specimen, thickness for cutting (ideal, do you use a punch like for skin biopsy)  and preparing the sausage (with or without wrapping , I've read that some people are using amnionic membrane), etc.  Do you retrieve the specimen wanted directly from the paraffin block then gathering all the specimen needed into the same cassette , etc.

 

As you can see I'm trying to get all the pertinent information before starting this project which will be very challenging into our institution.  

 

Thank you in advance for all the info you'll share

 

Have a nice day !

_________________________________________________________________
Cr?ez un personnage ? votre image pour votre WL Messenger
http://go.microsoft.com/?linkid=9656622
From resendes.ana <@t> gmail.com  Thu May 28 14:30:13 2009
From: resendes.ana <@t> gmail.com (Ana Resendes)
Date: Thu May 28 14:30:19 2009
Subject: [Histonet] immunophenotyping pigs
In-Reply-To: <3D8B70934FF500CE917904B9@CDYwxp1931.ad.med.buffalo.edu>
References: <905854.25089.qm@web50310.mail.re2.yahoo.com>
	<3D8B70934FF500CE917904B9@CDYwxp1931.ad.med.buffalo.edu>
Message-ID: 

For detailed information regarding this issue you should get incribed in the
vetimmunology forum.
Vetimm mailing list
Vetimm@vasci.umass.edu
https://list.umass.edu/mailman/listinfo/vetimm

there are plenty of papers published for pig immunophenotyping in the
journal of veterinary immunology and immunopathology
one good is
Chianini el al.


2009/5/21 Merced M Leiker 

> Anyone know how to immunophenotype pigs?  Or any published references?
> Thanks...
>
> Merced M Leiker
> Cardiovascular Medicine
> 348 Biomedical Research Building
> State University of New York at Buffalo
> 3435 Main St, Buffalo, NY  14214
> leiker@buffalo.edu
> 716-829-6118
>
> No trees were harmed in the sending of this email.
> However, many electrons were severely inconvenienced.
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>



-- 
Ana Resendes
DVM, MSc, PhD, Veterinary Pathologist
Postdoctoral DRCT-fellow research scientist
Centro de Investiga??o de Recursos Naturais (CIRN)
Laborat?rio de Microscopia e Histologia
Departamento de Biologia, Universidade dos A?ores.
Rua da M?e de Deus, 58 - Apartado 1422
P - 9501-801 Ponta Delgada (A?ores)
Portugal
Tel. (+351) 296 650 111
Fax (+351) 296 650 100
http://www.uac.pt/~pherg/
From resendes.ana <@t> gmail.com  Thu May 28 14:56:06 2009
From: resendes.ana <@t> gmail.com (Ana Resendes)
Date: Thu May 28 14:56:12 2009
Subject: [Histonet] microtome vibration:
Message-ID: 

In the lab I am currently working the microtome is often used to cut
speciments of hard shell snails and other mterials that most of the times
have calcifications that are difficult to retrieve, so most times tissues
have to go to further decalcification and I believe that this could have
caused an unvisable deviation on the microtome since, we have striations in
all parafin sections, even with new blades and soft tissues, that compromise
a good histology section for evaluation of tissues.
Anyone has any comments to do on a possible deviation a microtome can get.
Any suggestions or test to prove this possibility? If you were suspecting
from such a problem in your microtome, what would you do? contact the
manufecturer?
Anyone had this problem? Or is a deviation of the microtome possible when it
is used often with calcified tissues?

-- 
Ana Resendes
DVM, MSc, PhD, Veterinary Pathologist
Postdoctoral DRCT-fellow research scientist
Centro de Investiga??o de Recursos Naturais (CIRN)
Laborat?rio de Microscopia e Histologia
Departamento de Biologia, Universidade dos A?ores.
Rua da M?e de Deus, 58 - Apartado 1422
P - 9501-801 Ponta Delgada (A?ores)
Portugal
Tel. (+351) 296 650 111
Fax (+351) 296 650 100
http://www.uac.pt/~pherg/
From bliven.laura <@t> marshfieldclinic.org  Thu May 28 14:58:51 2009
From: bliven.laura <@t> marshfieldclinic.org (Bliven, Laura)
Date: Thu May 28 14:58:58 2009
Subject: [Histonet] Antibodies - nNOS & CD56
Message-ID: <200905281958.n4SJwqa1008831@mailhost2.mfldclin.edu>

I am looking for nNOS and CD56 antibodies that work well on formalin-fixed, paraffin embedded, human tissue. There are many CD56 clones. Also the monoclonal nNOS that we used didn't stain very well. Maybe I should be using a polyclonal?
Thanks for any input.
Laura



From rhbrown1 <@t> histocs.com  Thu May 28 14:55:16 2009
From: rhbrown1 <@t> histocs.com (Leroy Brown)
Date: Thu May 28 14:59:31 2009
Subject: [Histonet] looking for contact info on Dolby-Jamison
Message-ID: 

I am trying to contact Dolby  Jamison.   Does anyone have this info available?  

Thanks  

LeRoy Brown HT(ASCP) HTL  

histocs.com  

360-966-7300
From rjbuesa <@t> yahoo.com  Thu May 28 15:00:46 2009
From: rjbuesa <@t> yahoo.com (Rene J Buesa)
Date: Thu May 28 15:00:52 2009
Subject: [Histonet] Recycling Formalin for Fresh Tissue
Message-ID: <635261.73013.qm@web65705.mail.ac4.yahoo.com>

Recycling a known carcinogen like formalin, no matter the method, is something I would absolutely not do, nor encourage anybody to do.
One of the few advantages of formalin is that it is cheap and readily available.
It is absolutely not worth it to recycle it and get exposed more than needed.
Ren? J.

--- On Thu, 5/28/09, akemiat3377@yahoo.com  wrote:


From: akemiat3377@yahoo.com 
Subject: [Histonet] Recycling Formalin for Fresh Tissue
To: "Histonet" 
Date: Thursday, May 28, 2009, 12:14 PM


Good Morning Histo Land,

I am asking all you out there to give me your input on recycIing formalin.? I realize this has been discussed in the not too distant past, but this may be a little different situation.? I realize some labs are recycling formalin to put on their tissue processors, but this is a totally different scenario.

I have been a manager at my lab for only a month.? Much to my surprise, I just found out yesterday that our lab is recycling formalin and shipping it out to one of our clients to put their FRESH SPECIMENS into.? 

I just had a lengthy conversation with Robert Lott the day before yesterday, regarding pre-analytical protocol's effecting the test results of AFB.? By the way, we were in full agreement.? This formalin issue was unknown to me then.? I realize that the pre-analytical, pre-analytical, pre-analytical, process effects the final out come.? I need to present my case to our Director of Pathology why this is compromising patient care.

Any and all responses are gladly welcome.

Thanks,
Akemi

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287 
W: E-Mail: aallison-tacha@apmglab.com
P: E-Mail: akemiat3377@yahoo.com

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



      
From Robert.D.Kobus <@t> Medstar.net  Thu May 28 15:01:36 2009
From: Robert.D.Kobus <@t> Medstar.net (Robert.D.Kobus@Medstar.net)
Date: Thu May 28 15:01:48 2009
Subject: [Histonet] Robert D Kobus is out of the Laboratory.
Message-ID: 



I will be out of the office starting Wed 05/27/2009 and will not return
until Sun 05/31/2009.

Due to a death in the family, I will not be in the Laboratory till Monday
June 1, 2009.  While I am gone, please contact Michelle Gitu at 7-5282 or
in an emergency please call me on my cell at 540-470-0461.  When I return I
will respond to your message.

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From Paul <@t> Firnschild.com  Thu May 28 15:02:41 2009
From: Paul <@t> Firnschild.com (Paul Firnschild)
Date: Thu May 28 15:04:47 2009
Subject: [Histonet] help needed to find parts for tissue processor
Message-ID: <001901c9dfcf$41fddbb0$6402a8c0@qa44c0f386def9>

Leroy,

You could also have a electronics technician troubleshoot and repair your
screens.  You know. Transistors, diodes, triacs, capacitors which are all
available online. Hardly anyone even thinks of that option anymore.  Let me
know if I can help you.

Paul
 
Paul M. Firnschild
QA Support Services, Inc.
404.291.3715
 

-----Original Message-----
From: Leroy Brown [mailto:rhbrown1@histocs.com] 
Sent: Wednesday, May 27, 2009 12:24 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] help needed to find parts for tissue processor

Hi ,  I have two MVP-1 tissue processor  that are still in great shape
except that both have lost the display  screens.   Other than that they are
wonderful tissue  processors.    I am told by the folks that "know" this
machine that I can swap out these screens from any working model from  any
of the following processors.   The RMC models,  the MVP-1  or MVP-2 the
older 1530 , the Renaissance processor.   They are  all similar since this
machine changed hands so many times.  It was a  Fisher Scientific,  then
Instrumentation Lab,  then Vantana  and so on.   The problem is that the
parts are not available any  longer.    My only hope if I want to continue
using these  processors is to find a parts machine or one that is sitting in
the basement  of someones hospital or lab perhaps for years.  So, if you
have one or  know where to find one I would very much like to hear from you.
I bought these two processors about 2 years ago from a  government auction
still new in the ori
ginal crate  unopened.    I really would like to get them  up and running
again.   

Thanks  

LeRoy Brown HT(ASCP) HTL  

www.histocs.com  

email to me directly at lhbhcs@comcast.net or call me at  360-966-7300



From juditw <@t> u.washington.edu  Thu May 28 15:36:56 2009
From: juditw <@t> u.washington.edu (Judith L. Williams)
Date: Thu May 28 15:37:03 2009
Subject: [Histonet] sodium barbituate buffer cloudiness?
Message-ID: 


Hi everyone - need some fairly quick feedback. am making solutions for muscle enzyme histochem. the sodium barb. 0.1M buffer 2g in 100 mls is taking forever to mix, with heat and stirring. is this normal???? will it ever go clear?
Judy

Judith Williams, PhD, HT(ASCP)
Research Scientist
Department of Comparative Medicine
University of Washington
Seattle, WA 98195




From kgrobert <@t> rci.rutgers.edu  Thu May 28 16:06:47 2009
From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts)
Date: Thu May 28 15:53:11 2009
Subject: [Histonet] looking for contact info on Dolby-Jamison
In-Reply-To: <20090528200219.C7BBBABAA4@annwn22.rutgers.edu>
References: <20090528200219.C7BBBABAA4@annwn22.rutgers.edu>
Message-ID: <4A1EFCE7.6030309@rci.rutgers.edu>

>From www.dolbeyjamison.com:

Dolbey-Jamison provides product sales and service 
 for all of Pennsylvania, 
New Jersey, Maryland, Delaware and the New York City Metropolitan area. 













	

Phone

	

610.495.2010

Fax

	

610.495.2005

E-mail

	

sales@dolbey-jamison.com 


	

service@dolbey-jamison.com 

Address

	

213 Jones Boulevard - Suite 105
Pottstown, Pennsylvania, 19464



Leroy Brown wrote:

>I am trying to contact Dolby  Jamison.   Does anyone have this info available?  
>
>Thanks  
>
>LeRoy Brown HT(ASCP) HTL  
>
>histocs.com  
>
>360-966-7300
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>  
>
From billodonnell <@t> catholichealth.net  Thu May 28 16:02:01 2009
From: billodonnell <@t> catholichealth.net (O'Donnell, Bill)
Date: Thu May 28 16:02:10 2009
Subject: [Histonet] Multispecimen tissue blocks /sausage
In-Reply-To: 
References: 
Message-ID: 

 
Carmen,

I have developed a VERY inexpensive methodology for making tissue micro-array blocks for immuno controls. It is a bit time consuming, but no more than making sausage blocks. I would be happy to share it with you if you are interested. It conserves precious tissue and comfortably places 16+ pieces of tissue on a slide in about the space of a nickel, utilizing materials you very likely have in your lab.

William (Bill) O'Donnell, HT (ASCP) QIHC 
Lead Histologist
Good Samaritan Hospital
10 East 31st Street
Kearney, NE 68847


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of carmen loiselle
Sent: Thursday, May 28, 2009 2:23 PM
To: pathology education
Subject: [Histonet] Multispecimen tissue blocks /sausage


To all histonetters,

 

I need your expertise for the preparation of sausage multispecimen block for quality control in immunoshistochemistry lab.  At first my pathologist wanted to try microarray but because of the cost he had decided to go for sausage instead.  I would greatly appreciate any information regarding this matter starting with the procurement of the specimen, thickness for cutting (ideal, do you use a punch like for skin biopsy)  and preparing the sausage (with or without wrapping , I've read that some people are using amnionic membrane), etc.  Do you retrieve the specimen wanted directly from the paraffin block then gathering all the specimen needed into the same cassette , etc.

 

As you can see I'm trying to get all the pertinent information before starting this project which will be very challenging into our institution.  

 

Thank you in advance for all the info you'll share

 

Have a nice day !

_________________________________________________________________
Cr?ez un personnage ? votre image pour votre WL Messenger http://go.microsoft.com/?linkid=9656622_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


From jnocito <@t> satx.rr.com  Thu May 28 16:07:16 2009
From: jnocito <@t> satx.rr.com (Joe Nocito)
Date: Thu May 28 16:07:35 2009
Subject: [Histonet] archive help
Message-ID: 

Greetings all,
I'm going to admit that I'm a dummy and need help with getting info from the archives. I'm specifically looking for postings dealing with using freeze spray while doing frozens. So, how do I get to the archives again? That sucking noise is me pulling my head out of my butt. Thanks

JTT
From JWeems <@t> sjha.org  Thu May 28 16:09:50 2009
From: JWeems <@t> sjha.org (Weems, Joyce)
Date: Thu May 28 16:09:54 2009
Subject: [Histonet] archive help
In-Reply-To: 
References: 
Message-ID: <5D64396A0D4A5346BEBC759022AAEAA554CD8E@ITSSSXM01V6.one.ads.che.org>

http://www.histosearch.com/histonet.html


Vaseline is in the mail... J
 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe
Nocito
Sent: Thursday, May 28, 2009 5:07 PM
To: Histonet
Subject: [Histonet] archive help

Greetings all,
I'm going to admit that I'm a dummy and need help with getting info from
the archives. I'm specifically looking for postings dealing with using
freeze spray while doing frozens. So, how do I get to the archives
again? That sucking noise is me pulling my head out of my butt. Thanks

JTT
_______________________________________________
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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From macveigh <@t> usc.edu  Thu May 28 16:15:54 2009
From: macveigh <@t> usc.edu (Michelle MacVeigh-Aloni)
Date: Thu May 28 16:14:00 2009
Subject: [Histonet] microtome vibration:
Message-ID: <004e01c9dfd9$7c48c190$5c237d80@DFS66DD1>

Hi Ana,

First, I would remove the knife and the knife holder and clean everything from even the smallest debris.
After it is all spotless, I would put it together, make sure all screws are tight and try it again. More likely this will solve the problem.

Second - look carefully at the knife holder. We have had a problem with a notched knife holder that had to be polished patiently with fine sand paper for metal (on flat surface) until we removed the damage/notch from it and got it to be very smooth again.

Hope this helps
Michelle

USC Keck School of Medicine
From akemiat3377 <@t> yahoo.com  Thu May 28 16:46:54 2009
From: akemiat3377 <@t> yahoo.com (Akemi Allison-Tacha)
Date: Thu May 28 16:47:01 2009
Subject: Fw: Re: AW: [Histonet] AFB & Recycling Formalin for Fresh Tissue
Message-ID: <15928.69556.qm@web31306.mail.mud.yahoo.com>

Thanks Freida,

What better source to validate the steps I am taking to eliminate the problem of "AFB Contaminate".? 

I would also like to mention in my haste to compose my e-mail, I may have stated something incorrectly.? I did not mean that a positive (+) control be cut at the same time as the test sample and negative (-) control.? I usually place the test tissue on the same pre-cut positive (+) control slide.

Thanks Rene,

For your input on recycling formalin.? This is good ammunition for my defense in stopping the use of recycling formalin.? I still would welcome any and all replies.

Akemi


Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287 
W: E-Mail: aallison-tacha@apmglab.com
P: E-Mail: akemiat3377@yahoo.com



--- On Thu, 5/28/09, Freida Carson  wrote:

From: Freida Carson 
Subject: Re: Fw: Re: AW: [Histonet] AFB & Recycling Formalin for Fresh Tissue
To: "Histonet" , "Akemi Allison-Tacha" 
Date: Thursday, May 28, 2009, 12:09 PM

Wang reported on the contamination of tissue sections with AFB by the use of fluorescence microscopy, and?if I remember correctly, when he tested the water in water fountains, he found something like 33% contained AFB.? Non-pathogenic, but we can't tell that on our stains. The paper is in Am J Clin Pathol 51:71, 1969.? We (Carson, Kingsley, Haberman, and Race) also reported the contamination in the 1964 issue of the same journal.? We stopped using tap water for our water baths and began using millipore filtered water that had already been through a deionizing column and charcoal filter.? We always cut a negative control from the same days workload as the patient case - uterus in our case.? You don't need a positive control cut the same day, just a positive control with only a medium?number of organisms. ?We
 also?did not use any tap water in the deparaffinization prior to the carbol fuchsin.? AFB organisms have also been reported growing in 40 gal formalin tanks believe it or not and in the old?Fisher?Paraffin wafers.
?
We centrifuged?tap water in 50-ml tubes, poured off the supernatent, and refilled and recentrifuged until we could see some sediment in the bottom.? We took that to Microbioloby and had it cultured to definited prove the presence of AFB in the tap water.
I?would recommend this approach to anyone suspecting?tissue contamination.?
?
Freida Carson
--- On Thu, 5/28/09, Akemi Allison-Tacha  wrote:


From: Akemi Allison-Tacha 
Subject: Fw: Re: AW: [Histonet] AFB & Recycling Formalin for Fresh Tissue
To: "Histonet" 
Date: Thursday, May 28, 2009, 1:21 PM


Hello,
I responded to Gudrun personally, but I thought that the information I am supplying him below might be a good educational source for other histologists who are having similar scenario's for AFB contamination.? 

I would also like to say that I have an extremely eager histology team that have expressed an interest in learning theory and practice.? Most of my team are OJT.? They have had several pathologists approach them with these concerns regarding AFB, and because of their knowledge, or lack of it, couldn't fix the problem.? We are starting with the basics and I am giving in-services along the way.

Hi? Gudrun,

They are shipping the recycled formalin directly to the outside hospital surgery to put their fresh tissues into.....

As far as the AFB, we have had intermitant (+) AFB on the edges of tissues, that has been extremely questionable.? We currently are not
 cutting a negative (-) & positive(+) control on the same water bath as the test sample. That will be remedied beginning Monday. The water here is also in question.? I will be sending it out for microbacteria analysis.

We are going through a thorough housekeeping and in-service process.? We are starting with the tissue processors, which are currently only changed weekly (not rotated daily), embedding centers, which have to my knowledge, never been drained, and the hopper cleaned-out.? The water baths are not scrubbed out with soap and HOT water daily and
covered at night to prevent contamination.? The water bath is not skimmed of excess tissue debris, each and every time another specimen is placed on it.? The Carbol Fuchsin is not being filtered prior to use.? The working solution is currently being poured back into the stock bottle.? Need I go on................

Akemi Allison-Tacha BS, HT (ASCP)
 HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287 
W: E-Mail: aallison-tacha@apmglab.com
P: E-Mail: akemiat3377@yahoo.com



--- On Thu, 5/28/09, Gudrun Lang  wrote:

From: Gudrun Lang 
Subject: AW: [Histonet] Recycling Formalin for Fresh Tissue
To: akemiat3377@yahoo.com
Cc:
histonet@lists.utsouthwestern.edu
Date: Thursday, May 28, 2009, 10:34 AM

Just for interest. Your lab takes the recycled formalin to make 4% neutral
buffered formalin (NBF)? Or do they ship the recycled formalin directly in
bottles to the surgery?
AFB = acid fast bacili?

Do you have concerns, that they produce false-positive AFB stains? I cannot
imagine, that a (hopefully) small number of dead bacili on the surface of
tissue will pretend this. An other question: Isn't there a filter in the
recycler to prevent such contamination?
Gudrun

-----Urspr?ngliche Nachricht-----
Von: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von
akemiat3377@yahoo.com
Gesendet: Donnerstag, 28. Mai 2009 18:14
An: Histonet
Betreff: [Histonet] Recycling Formalin for Fresh Tissue

Good Morning Histo Land,

I am asking all you out there to give me your input on recycIing formalin.?
I realize this has been discussed in the not too distant past, but this may
be a little different situation.? I realize some labs are recycling formalin
to put on their tissue processors, but this is a totally different
 scenario.

I have been a manager at my lab for only a month.? Much to my surprise, I
just found out yesterday that our lab is recycling formalin and shipping it
out to one of our clients to put their FRESH SPECIMENS into.?


I just had a lengthy conversation with Robert Lott the day before yesterday,
regarding pre-analytical protocol's effecting the test results of AFB.? By
the way, we were in full agreement.? This formalin issue was unknown to me
then.? I realize that the pre-analytical, pre-analytical, pre-analytical,
process effects the final out come.? I need to present my case to our
Director of Pathology why this is compromising patient care.

Any and all responses are gladly welcome.

Thanks,
Akemi

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Ct. Los Gatos, CA 95032
Cell:
 425.941.4287 
W: E-Mail: aallison-tacha@apmglab.com
P: E-Mail: akemiat3377@yahoo.com

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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From algranth <@t> email.arizona.edu  Thu May 28 17:08:25 2009
From: algranth <@t> email.arizona.edu (Andrea Grantham)
Date: Thu May 28 17:08:30 2009
Subject: [Histonet] Advice on fatty liver sectioning
In-Reply-To: 
References: 
Message-ID: 

-18? is the temp I use when I section liver for ORO and I usually cut  
the sections at 5 microns. It sounds like your tissue was not frozen  
correctly.

Andi



Andrea Grantham, HT (ASCP)
Senior Research Specialist
University of Arizona
Cell Biology and Anatomy
Histology Service Laboratory
P.O.Box 245044
Tucson, AZ 85724

algranth@email.arizona.edu
Tel: 520.626.4415     Fax: 520.626.2097

"happy slicing and dicing and may all your stains work perfectly" -  
Paula Sicurello
P Please consider the environment before printing this email.



On May 28, 2009, at 12:20 PM, Kinsley, David wrote:

> Hi,
> I have some very fatty mouse livers that I have to obtain frozen  
> sections for Oil red O and IHC.  I have tried sectioning from -18 to  
> -30 degrees and from 5um to 10um and the liver just seems to  
> compress, pull away from the OCT, or roll up on itself while the OCT  
> sections well.  When I section below -30 parts of the liver have  
> chatter in them as if they are too cold.  Does anyone have any  
> suggestions on what the best cutting temperature would be for these?
>
> I have successfully cut normal and less fatty livers at 5um ad 10 um  
> at -18 and -25 degrees respectively, it is these extremely fatty  
> livers that are giving me problems.  Any advice is appreciated.
>
> Dave
>
>
>
>
>
> *********************************************************************
> This message and any attachments are solely for the
> intended recipient. If you are not the intended recipient,
> disclosure, copying, use or distribution of the information
> included in this message is prohibited -- Please
> immediately and permanently delete.
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>

From crochieresteve <@t> aol.com  Thu May 28 18:17:57 2009
From: crochieresteve <@t> aol.com (crochieresteve@aol.com)
Date: Thu May 28 18:18:10 2009
Subject: [Histonet] Histology Supervisor opening in Springfield MA
Message-ID: <8CBAE01ABE06A12-39C-A8E@webmail-dd21.sysops.aol.com>

LifePath Partners, LLC has an immediate opening for a supervisory level histotechnologist with experience in IHC to oversee daily operations of the main histology lab at Mercy Medical Center and the satellite prostate bx labs at Urology Group of Western New England. Staff includes 4 FTE Histotechs and 2 FTE Laboratory assistants. Annual surgical volume is 25k +/-.
If interested send resume or call:

Christine Sullivan, Business Manager
Christine.sullivan@sphs.com
299 Carew St.
Springfield, MA 01104
(413) 748-9779
From CIngles <@t> uwhealth.org  Thu May 28 18:40:17 2009
From: CIngles <@t> uwhealth.org (Ingles Claire )
Date: Thu May 28 18:43:05 2009
Subject: [Histonet] Horseshoe crab..help!
References: <8ED3F2CA5B78E142B8193376C57330F8E198E18F55@EXCH2007.ad.stir.ac.uk>
Message-ID: 

Anyone please correct me if I'm wrong, but is chitin similar to fingernails? If so, we just use Nair for a bit. You may need to use it for a few hours since the shell in horseshoe crabs is harder than regular fingernails.
Claire

________________________________

From: histonet-bounces@lists.utsouthwestern.edu on behalf of Deborah Faichney
Sent: Thu 5/28/2009 4:56 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Horseshoe crab..help!



Hi all,

I have a Masters student here who has Horseshoe crab tissues fixed in 10% NBF.  Most of the tissues have been sectioned successfully but the eyes are surrounded by armour plated chitin! (she broke two Dremel drills trying to cut the carapace and finally tin snips had to be used to cut them out!!).
Any thoughts on how to soften them?  Looking around on the Net, I have found  that processing and clearing through Chloroform may soften, also a solution called Diaphanol has been recommended.
At the moment they have been two days in a 1 part acetic acid to 5 parts NBF with no effect.
We only have a couple of weeks to do this work so cannot adopt any trial or lengthy methods.

Is this possible?  I like a challenge, but suspect this needs a miracle.  Its no wonder these beasties have been on the planet for so long.
Many thanks

Debbie Faichney
Histopathology
Institute of Aquaculture
University of Stirling
Stirling, FK7 7QS
Scotland
Uk




--
Academic Excellence at the Heart of Scotland.
The University of Stirling is a charity registered in Scotland,
 number SC 011159.

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From lscott <@t> sfcn.org  Thu May 28 23:42:12 2009
From: lscott <@t> sfcn.org (Scott)
Date: Thu May 28 23:42:34 2009
Subject: [Histonet] Sakura pigma micro 01 pens
Message-ID: <0B5BDFD9DD1144DE8E1101C5E8498634@LesliePC>

Hi,
    Does anyone use the Sakura pigma micron 01 pens?  Our supplier no longer sales them.  If so I would like to get the supplier info, Thanks!



Scott Hendricksen, HT(ASCP)
From lscott <@t> sfcn.org  Fri May 29 00:03:16 2009
From: lscott <@t> sfcn.org (Scott)
Date: Fri May 29 00:03:35 2009
Subject: [Histonet] Sakura film coverslipper xylene residue issue
Message-ID: <36FFE50816914030A16AC7213B94A1E2@LesliePC>

Hi,
    We have a newer Sakura film coverslipper, does anyone have experience with this piece of equipment. I have a problem with the slides having a light xylene residue, after the slides come off of the coverslipper. 

How do you dry them? If I leave the slides on the coverslipper to air dry the xylene residue is noticeable. If I lightly wipe off all of the slides as they come off the coverslipper they look clean.

How do you guys do it? I think it is a waste of time to wipe off all of the slides. I don't think the xylene flow is too high.

Thanks for your help,

Scott Hendricksen HT(ASCP)
From Kemlo.Rogerson <@t> waht.swest.nhs.uk  Fri May 29 04:15:05 2009
From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson)
Date: Fri May 29 04:15:11 2009
Subject: [Histonet] Recycling Formalin for Fresh Tissue
Message-ID: <86ADE4EB583CE64799A9924684A0FBBF06E12F99@wahtntex2.waht.swest.nhs.uk>

Why would you want to? Surely the time and energy needed to make sure that the recycled formalin was of the correct strength, was not infective and was at the correct pH and properly buffered would not make it cost effective.

Bit like using a teabag twice; that's uncivilised!!!


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of akemiat3377@yahoo.com
Sent: 28 May 2009 17:14
To: Histonet
Subject: [Histonet] Recycling Formalin for Fresh Tissue

Good Morning Histo Land,

I am asking all you out there to give me your input on recycIing formalin.? I realize this has been discussed in the not too distant past, but this may be a little different situation.? I realize some labs are recycling formalin to put on their tissue processors, but this is a totally different scenario.

I have been a manager at my lab for only a month.? Much to my surprise, I just found out yesterday that our lab is recycling formalin and shipping it out to one of our clients to put their FRESH SPECIMENS into.? 

I just had a lengthy conversation with Robert Lott the day before yesterday, regarding pre-analytical protocol's effecting the test results of AFB.? By the way, we were in full agreement.? This formalin issue was unknown to me then.? I realize that the pre-analytical, pre-analytical, pre-analytical, process effects the final out come.? I need to present my case to our Director of Pathology why this is compromising patient care.

Any and all responses are gladly welcome.

Thanks,
Akemi

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories 105A Cooper Ct. Los Gatos, CA 95032
Cell: 425.941.4287
W: E-Mail: aallison-tacha@apmglab.com
P: E-Mail: akemiat3377@yahoo.com

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

From lpwenk <@t> sbcglobal.net  Fri May 29 05:00:04 2009
From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk)
Date: Fri May 29 05:00:14 2009
Subject: [Histonet] archive help
In-Reply-To: 
Message-ID: <05252C55381A4598821220ABD38DC03A@HPPav2>


CFR 1910.1030  (OSHA regulation)

"All procedures involving blood or other potential infectious materials
shall be performed in such as manner as to minimize splashing, spraying,
spattering, or generation of droplets of these substances."


CLSI DOCUMENT M29  (Clinical Laboratory Standards Institute (what CAP is
always quoting))

- Protection of Laboratory Workers

"Frozen sections done on unfixed tissue pose a high risk because accidents
are common.  Freezing of tissue does not inactivate infectious agents.
Freezing propellants under pressure should not be used for frozen sections
as they may cause spattering of droplets of infectious material."
 
Peggy A. Wenk, HTL(ASCP)SLS
Beaumont Hospital
Royal Oak, MI 48073

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito
Sent: Thursday, May 28, 2009 5:07 PM
To: Histonet
Subject: [Histonet] archive help

Greetings all,
I'm going to admit that I'm a dummy and need help with getting info from the
archives. I'm specifically looking for postings dealing with using freeze
spray while doing frozens. So, how do I get to the archives again? That
sucking noise is me pulling my head out of my butt. Thanks

JTT
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From njoydobro <@t> aol.com  Fri May 29 06:19:30 2009
From: njoydobro <@t> aol.com (njoydobro@aol.com)
Date: Fri May 29 06:19:45 2009
Subject: [Histonet] Peroxidase block again................
Message-ID: <8CBAE66781E438A-38C-1C80@FWM-M20.sysops.aol.com>

good morning,

???? I would like to pose this questions again...Is there anyone out there who has eliminated using Peroxidase Block on either IHC or ICC?
Your feedback is very helpful.


Thanks,
Gene
Cleveland Clinic
From rjbuesa <@t> yahoo.com  Fri May 29 07:52:14 2009
From: rjbuesa <@t> yahoo.com (Rene J Buesa)
Date: Fri May 29 07:52:19 2009
Subject: [Histonet] Sakura film coverslipper xylene residue issue
Message-ID: <260661.66606.qm@web65708.mail.ac4.yahoo.com>

The solution is not drying the slides but regulating the amount of xylene dispensed over each section before the film rolls over than, in spite of what you think, has to be excessive.
Being a new instrument I think that you are better off by asking the Sakura sales person.
Ren? J.

--- On Fri, 5/29/09, Scott  wrote:


From: Scott 
Subject: [Histonet] Sakura film coverslipper xylene residue issue
To: Histonet@lists.utsouthwestern.edu
Date: Friday, May 29, 2009, 1:03 AM


Hi,
? ? We have a newer Sakura film coverslipper, does anyone have experience with this piece of equipment. I have a problem with the slides having a light xylene residue, after the slides come off of the coverslipper. 

How do you dry them? If I leave the slides on the coverslipper to air dry the xylene residue is noticeable. If I lightly wipe off all of the slides as they come off the coverslipper they look clean.

How do you guys do it? I think it is a waste of time to wipe off all of the slides. I don't think the xylene flow is too high.

Thanks for your help,

Scott Hendricksen HT(ASCP)
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



      
From carmenliza_stefan <@t> yahoo.com  Fri May 29 07:54:18 2009
From: carmenliza_stefan <@t> yahoo.com (carmen stefan)
Date: Fri May 29 07:54:23 2009
Subject: [Histonet] Sakura film coverslipper xylene residue issue
Message-ID: <223563.64207.qm@web32207.mail.mud.yahoo.com>

Hi Scott
We noticed the same problem, but we decided to dry the slides under the fome hood (where the air flow is stronger and the process is faster). The slides are ok, we might have some with xylene, but we clean, at the end, only those.



      
From anh2006 <@t> med.cornell.edu  Fri May 29 08:18:38 2009
From: anh2006 <@t> med.cornell.edu (Andrea Hooper)
Date: Fri May 29 08:18:47 2009
Subject: [Histonet] H&E
Message-ID: 

POLL: What is everyone's favorite hematoxylin for work-horse H&E 
stains? What about eosin?
I am re-evaluating the quality of our current set-up and think we can 
improve - especially for the frozen sections.
Any suggestions/protocols/brand suggestions?

Thanks,
Andrea
-- 

From KGroeger <@t> USLABS.net  Fri May 29 08:19:22 2009
From: KGroeger <@t> USLABS.net (Karin Groeger)
Date: Fri May 29 08:19:28 2009
Subject: [Histonet] Sakura film coverslipper xylene residue issue
In-Reply-To: <36FFE50816914030A16AC7213B94A1E2@LesliePC>
References: <36FFE50816914030A16AC7213B94A1E2@LesliePC>
Message-ID: 

Hi Scott,
  We place ours in front of a fan to dry if we are in a hurry, even if
we air dry them we do not have a problem with the xylene residue,
perhaps you have  a bad batch, check your lot # and call Sakura.  We
coverslip over 1,000 slides a day and have not seen this.

Karin Groeger

Histology Supervisor

US LABS, Irvine,CA

949-450-0145 ext. 649

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Scott
Sent: Thursday, May 28, 2009 10:03 PM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Sakura film coverslipper xylene residue issue

Hi,
    We have a newer Sakura film coverslipper, does anyone have
experience with this piece of equipment. I have a problem with the
slides having a light xylene residue, after the slides come off of the
coverslipper. 

How do you dry them? If I leave the slides on the coverslipper to air
dry the xylene residue is noticeable. If I lightly wipe off all of the
slides as they come off the coverslipper they look clean.

How do you guys do it? I think it is a waste of time to wipe off all of
the slides. I don't think the xylene flow is too high.

Thanks for your help,

Scott Hendricksen HT(ASCP)
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



Confidentiality Notice: This message, including any attachments, may contain CONFIDENTIAL AND/OR LEGALLY PRIVILEGED information. The information is intended for use by the individual named above and may not be disseminated to any other party without US LABS' written permission. If you are not the intended recipient, or the employee or agent responsible for delivering this information to the intended recipient, you are hereby notified that any dissemination, disclosure, distribution, copying or taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this information in error, please notify US LABS immediately at 1-888-450-0145 attn: Compliance Department to arrange for return of this message including all attachments.

From mpence <@t> grhs.net  Fri May 29 08:19:51 2009
From: mpence <@t> grhs.net (Mike Pence)
Date: Fri May 29 08:19:56 2009
Subject: [Histonet] Sakura film coverslipper xylene residue issue
In-Reply-To: <36FFE50816914030A16AC7213B94A1E2@LesliePC>
Message-ID: <661949901A768E4F9CC16D8AF8F2838C017A3B61@IS-E2K3.grhs.net>

I have used this coverslipper for over 10 years and what I have found is
to allow the slides to dry under a fume hood in the rack as they come
off the coverslipper for 5 minutes. This allows the excess xylene to run
to the frosted end of the slide.

Also, the correct amount for dispensing xylene on the slide is a "string
of pearls" drop placement. This leaves about 5mm between drops from
beginning to end of slide.

Mike

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Scott
Sent: Friday, May 29, 2009 12:03 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Sakura film coverslipper xylene residue issue


Hi,
    We have a newer Sakura film coverslipper, does anyone have
experience with this piece of equipment. I have a problem with the
slides having a light xylene residue, after the slides come off of the
coverslipper. 

How do you dry them? If I leave the slides on the coverslipper to air
dry the xylene residue is noticeable. If I lightly wipe off all of the
slides as they come off the coverslipper they look clean.

How do you guys do it? I think it is a waste of time to wipe off all of
the slides. I don't think the xylene flow is too high.

Thanks for your help,

Scott Hendricksen HT(ASCP)
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



From robert_schoonhoven <@t> yahoo.com  Fri May 29 08:35:42 2009
From: robert_schoonhoven <@t> yahoo.com (Robert Schoonhoven)
Date: Fri May 29 08:35:45 2009
Subject: [Histonet] looking for Ford Royer
Message-ID: <765914.66157.qm@web31101.mail.mud.yahoo.com>

  Please contact me off list.

Thanks

 Robert Schoonhoven, HT/HTL (ASCP)



      
From annigyg <@t> gmail.com  Fri May 29 08:36:01 2009
From: annigyg <@t> gmail.com (Anne van Binsbergen)
Date: Fri May 29 08:36:08 2009
Subject: [Histonet] Multispecimen tissue blocks /sausage
In-Reply-To: 
References: 
	
Message-ID: 

i am very interested in your technique - please do share here on the
histonet if possible
thanks
Annie in Abu Dhabi

2009/5/29 O'Donnell, Bill 

>
> Carmen,
>
> I have developed a VERY inexpensive methodology for making tissue
> micro-array blocks for immuno controls. It is a bit time consuming, but no
> more than making sausage blocks. I would be happy to share it with you if
> you are interested. It conserves precious tissue and comfortably places 16+
> pieces of tissue on a slide in about the space of a nickel, utilizing
> materials you very likely have in your lab.
>
> William (Bill) O'Donnell, HT (ASCP) QIHC
> Lead Histologist
> Good Samaritan Hospital
> 10 East 31st Street
> Kearney, NE 68847
>
>
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu [mailto:
> histonet-bounces@lists.utsouthwestern.edu] On Behalf Of carmen loiselle
> Sent: Thursday, May 28, 2009 2:23 PM
> To: pathology education
> Subject: [Histonet] Multispecimen tissue blocks /sausage
>
>
> To all histonetters,
>
>
>
> I need your expertise for the preparation of sausage multispecimen block
> for quality control in immunoshistochemistry lab.  At first my pathologist
> wanted to try microarray but because of the cost he had decided to go for
> sausage instead.  I would greatly appreciate any information regarding this
> matter starting with the procurement of the specimen, thickness for cutting
> (ideal, do you use a punch like for skin biopsy)  and preparing the sausage
> (with or without wrapping , I've read that some people are using amnionic
> membrane), etc.  Do you retrieve the specimen wanted directly from the
> paraffin block then gathering all the specimen needed into the same cassette
> , etc.
>
>
>
> As you can see I'm trying to get all the pertinent information before
> starting this project which will be very challenging into our institution.
>
>
>
> Thank you in advance for all the info you'll share
>
>
>
> Have a nice day !
>
> _________________________________________________________________
> Cr?ez un personnage ? votre image pour votre WL Messenger
> http://go.microsoft.com/?linkid=9656622_______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>



-- 
Anne van Binsbergen (Hope)
Abu Dhabi
UAE
From Jackie.O'Connor <@t> abbott.com  Fri May 29 08:52:27 2009
From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor)
Date: Fri May 29 08:52:46 2009
Subject: [Histonet] Sakura film coverslipper xylene residue issue
In-Reply-To: 
Message-ID: 

I have an older Sakura coverslipper, circa 1997ish.  We have noticed an 
increasing number of brown spots on tissue sections recently.   This does 
not occur if the same slides are covered with glass coverslips.   It does 
not look like a drying artefact, occurs randomly throughout tissue types, 
location on tissue, i.e., edge, center.   I've even showed these spots to 
the rep who admitted she has never seen this artefact before.   We use 
only the Tissue Tek tape, we maintain the equipment on a regular basis. 
The xylene flow is good.   Our only problem are these random brown spots.  
 Any suggestions?
Jackie O'



"Karin Groeger"  
Sent by: histonet-bounces@lists.utsouthwestern.edu
05/29/2009 08:19 AM

To
"Scott" , 
cc

Subject
RE: [Histonet] Sakura film coverslipper xylene residue issue






Hi Scott,
  We place ours in front of a fan to dry if we are in a hurry, even if
we air dry them we do not have a problem with the xylene residue,
perhaps you have  a bad batch, check your lot # and call Sakura.  We
coverslip over 1,000 slides a day and have not seen this.

Karin Groeger

Histology Supervisor

US LABS, Irvine,CA

949-450-0145 ext. 649

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Scott
Sent: Thursday, May 28, 2009 10:03 PM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Sakura film coverslipper xylene residue issue

Hi,
    We have a newer Sakura film coverslipper, does anyone have
experience with this piece of equipment. I have a problem with the
slides having a light xylene residue, after the slides come off of the
coverslipper. 

How do you dry them? If I leave the slides on the coverslipper to air
dry the xylene residue is noticeable. If I lightly wipe off all of the
slides as they come off the coverslipper they look clean.

How do you guys do it? I think it is a waste of time to wipe off all of
the slides. I don't think the xylene flow is too high.

Thanks for your help,

Scott Hendricksen HT(ASCP)
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



Confidentiality Notice: This message, including any attachments, may 
contain CONFIDENTIAL AND/OR LEGALLY PRIVILEGED information. The 
information is intended for use by the individual named above and may not 
be disseminated to any other party without US LABS' written permission. If 
you are not the intended recipient, or the employee or agent responsible 
for delivering this information to the intended recipient, you are hereby 
notified that any dissemination, disclosure, distribution, copying or 
taking of any action in reliance on the contents of this information is 
strictly prohibited. If you have received this information in error, 
please notify US LABS immediately at 1-888-450-0145 attn: Compliance 
Department to arrange for return of this message including all 
attachments.

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From laurie.colbert <@t> huntingtonhospital.com  Fri May 29 09:07:31 2009
From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert)
Date: Fri May 29 09:07:36 2009
Subject: [Histonet] Sakura film coverslipper xylene residue issue
Message-ID: <57BE698966D5C54EAE8612E8941D768305AE3038@EXCHANGE3.huntingtonhospital.com>

We set the racks in front of a small fan.  It would be nice if Sakura
could incorporate some kind of drying fan in their coverslipper.
Laurie Colbert

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Scott
Sent: Thursday, May 28, 2009 10:03 PM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Sakura film coverslipper xylene residue issue

Hi,
    We have a newer Sakura film coverslipper, does anyone have
experience with this piece of equipment. I have a problem with the
slides having a light xylene residue, after the slides come off of the
coverslipper. 

How do you dry them? If I leave the slides on the coverslipper to air
dry the xylene residue is noticeable. If I lightly wipe off all of the
slides as they come off the coverslipper they look clean.

How do you guys do it? I think it is a waste of time to wipe off all of
the slides. I don't think the xylene flow is too high.

Thanks for your help,

Scott Hendricksen HT(ASCP)
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

From vetlab <@t> caribsurf.com  Fri May 29 09:11:54 2009
From: vetlab <@t> caribsurf.com (Veterinary Services Laboratory)
Date: Fri May 29 09:12:23 2009
Subject: [Histonet] Microtome Problems
Message-ID: 

 

Hello All

 

I'm new to the group and histo. (3 years) so far I'm getting a lot of useful
tips from all the messages.  Thank you.

 

I have two microtomes that were working almost perfectly before I had a
technician service them.  Now, the Leica 2035 Jung biocuts' blade holder is
twisting to the side (going up at one end).  I have tried tightening,
slacking, and cleaning the entire assembly to no avail.  Does anyone know of
where I might be able to source one or even how to fix the one I have.

 

Secondly, the Leitz 1512 rotary microtome, is now jumping forward and giving
a loud clanking noise.  It takes huge chunks out of the tissue.

 

Any and all help is appreciated,

 

Thank you

Sharon

 

Ms. Sharon Drayton

Veterinary Services Laboratory

Ministry of Agriculture & Rural Development

The Pine

St. Michael BB11091

Barbados

 

Tel:  (246) 427-5492 or (246) 427-5073

Fax: (246)426-7517

Email:  vetlab@caribsurf.com

Website:  www.agriculture.gov.bb

 

 

From lpaveli1 <@t> hurleymc.com  Fri May 29 09:13:11 2009
From: lpaveli1 <@t> hurleymc.com (Lynette Pavelich)
Date: Fri May 29 09:13:25 2009
Subject: [Histonet] Sakura film coverslipper xylene residue issue
Message-ID: <4A1FB537020000EE0002989C@smtp-gw.hurleymc.com>

Our Sakura coverslipper is from 1994 (yep...I win the prize!!).  When we
have had those "brown" spots, we have discovered that they are actually
dry areas.  I would look at the last little white roller bar on the
track.  The springs(2) that keeps it in place, and keeps that roller bar
nice and flat to squish out the air bubbles, may be going bad.  When we
have replaced the springs, our brown spots have been eliminated.  
In addition, we have to have the volume of xylene at about 6 drops to
accomadate the cytology smears, which does leave the slides a little on
the wet side.  So we just put them under the hood for a couple minutes
to dry.

Hope this helped!  Happy Friday!!!
Lynette

>>> Jackie M O'Connor  05/29/09 9:52 AM >>>
I have an older Sakura coverslipper, circa 1997ish.  We have noticed an 
increasing number of brown spots on tissue sections recently.   This
does 
not occur if the same slides are covered with glass coverslips.   It
does 
not look like a drying artefact, occurs randomly throughout tissue
types, 
location on tissue, i.e., edge, center.   I've even showed these spots
to 
the rep who admitted she has never seen this artefact before.   We use 
only the Tissue Tek tape, we maintain the equipment on a regular basis. 
The xylene flow is good.   Our only problem are these random brown
spots.  
 Any suggestions?
Jackie O'



"Karin Groeger"  
Sent by: histonet-bounces@lists.utsouthwestern.edu
05/29/2009 08:19 AM

To
"Scott" , 
cc

Subject
RE: [Histonet] Sakura film coverslipper xylene residue issue






Hi Scott,
  We place ours in front of a fan to dry if we are in a hurry, even if
we air dry them we do not have a problem with the xylene residue,
perhaps you have  a bad batch, check your lot # and call Sakura.  We
coverslip over 1,000 slides a day and have not seen this.

Karin Groeger

Histology Supervisor

US LABS, Irvine,CA

949-450-0145 ext. 649

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Scott
Sent: Thursday, May 28, 2009 10:03 PM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Sakura film coverslipper xylene residue issue

Hi,
    We have a newer Sakura film coverslipper, does anyone have
experience with this piece of equipment. I have a problem with the
slides having a light xylene residue, after the slides come off of the
coverslipper. 

How do you dry them? If I leave the slides on the coverslipper to air
dry the xylene residue is noticeable. If I lightly wipe off all of the
slides as they come off the coverslipper they look clean.

How do you guys do it? I think it is a waste of time to wipe off all of
the slides. I don't think the xylene flow is too high.

Thanks for your help,

Scott Hendricksen HT(ASCP)
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Confidentiality Notice: This message, including any attachments, may 
contain CONFIDENTIAL AND/OR LEGALLY PRIVILEGED information. The 
information is intended for use by the individual named above and may
not 
be disseminated to any other party without US LABS' written permission.
If 
you are not the intended recipient, or the employee or agent responsible

for delivering this information to the intended recipient, you are
hereby 
notified that any dissemination, disclosure, distribution, copying or 
taking of any action in reliance on the contents of this information is 
strictly prohibited. If you have received this information in error, 
please notify US LABS immediately at 1-888-450-0145 attn: Compliance 
Department to arrange for return of this message including all 
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From jeanie.meyers <@t> trumbulllabs.com  Fri May 29 09:26:38 2009
From: jeanie.meyers <@t> trumbulllabs.com (Jeanie Meyers)
Date: Fri May 29 09:23:41 2009
Subject: [Histonet] (no subject)
Message-ID: <200905291423.n4TENbFx029002@mail40.atl.registeredsite.com>

Please remove me from the listserve.

 

Jeanie Meyers, Laboratory Manager

 

Trumbull Laboratories, LLC

7550 Wolf River Boulevard, Suite 200

Germantown, Tennessee 38138

 

(901)542-6806    (901)507-2633 Fax

 

 

 

 

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 joseph-galbraith <@t> uiowa.edu  Fri May 29 09:35:22 2009
From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe)
Date: Fri May 29 09:35:27 2009
Subject: [Histonet] Microtome Problems
In-Reply-To: 
References: 
Message-ID: 

Sharon:

I will let others with personal experience with your particular
microtomes offer advice on how to repair them but, first and foremost,
if these problems occurred as a direct result of the service call, I
would be in immediate contact with the company that provided the
service, complain to them about the outcome you experienced, and insist
that they make the situation right by you.  Especially if you contact
them right away, they should at the least return to correct the problems
they created, possibly by sending a different rep.  Best of luck.

Joe

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Veterinary Services Laboratory
Sent: Friday, May 29, 2009 9:12 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Microtome Problems


 

Hello All

 

I'm new to the group and histo. (3 years) so far I'm getting a lot of
useful
tips from all the messages.  Thank you.

 

I have two microtomes that were working almost perfectly before I had a
technician service them.  Now, the Leica 2035 Jung biocuts' blade holder
is
twisting to the side (going up at one end).  I have tried tightening,
slacking, and cleaning the entire assembly to no avail.  Does anyone
know of
where I might be able to source one or even how to fix the one I have.

 

Secondly, the Leitz 1512 rotary microtome, is now jumping forward and
giving
a loud clanking noise.  It takes huge chunks out of the tissue.

 

Any and all help is appreciated,

 

Thank you

Sharon

 

Ms. Sharon Drayton

Veterinary Services Laboratory

Ministry of Agriculture & Rural Development

The Pine

St. Michael BB11091

Barbados

 

Tel:  (246) 427-5492 or (246) 427-5073

Fax: (246)426-7517

Email:  vetlab@caribsurf.com

Website:  www.agriculture.gov.bb

 

 

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

From trathborne <@t> somerset-healthcare.com  Fri May 29 09:38:26 2009
From: trathborne <@t> somerset-healthcare.com (Rathborne, Toni)
Date: Fri May 29 09:38:36 2009
Subject: [Histonet] Microtome Problems
In-Reply-To: 
Message-ID: 

Usually the service techs that come in will give a 30 day call-back period that is included as part of the service call. I would have them come back until it is working properly.

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of
Veterinary Services Laboratory
Sent: Friday, May 29, 2009 10:12 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Microtome Problems


 

Hello All

 

I'm new to the group and histo. (3 years) so far I'm getting a lot of useful
tips from all the messages.  Thank you.

 

I have two microtomes that were working almost perfectly before I had a
technician service them.  Now, the Leica 2035 Jung biocuts' blade holder is
twisting to the side (going up at one end).  I have tried tightening,
slacking, and cleaning the entire assembly to no avail.  Does anyone know of
where I might be able to source one or even how to fix the one I have.

 

Secondly, the Leitz 1512 rotary microtome, is now jumping forward and giving
a loud clanking noise.  It takes huge chunks out of the tissue.

 

Any and all help is appreciated,

 

Thank you

Sharon

 

Ms. Sharon Drayton

Veterinary Services Laboratory

Ministry of Agriculture & Rural Development

The Pine

St. Michael BB11091

Barbados

 

Tel:  (246) 427-5492 or (246) 427-5073

Fax: (246)426-7517

Email:  vetlab@caribsurf.com

Website:  www.agriculture.gov.bb

 

 

_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


CONFIDENTIALITY NOTICE
This message and any included attachments are from 
Somerset Medical Center and are intended only for the addressee.
The information contained in this message is confidential 
and may contain privileged, confidential, proprietary and/or 
trade secret information entitled to protection and/or exemption 
from disclosure under applicable law. Unauthorized forwarding, 
printing, copying, distribution, or use of such information is 
strictly prohibited and may be unlawful. If you are not the 
addressee, please promptly delete this message and notify 
the sender of the delivery error by e-mail or you may call 
Somerset Medical Center's computer Help Desk at 
908-685-2200, ext. 4050. 
-------------------------------------------------------------- 
Somerset Medical Center is the recipient of the 
2009 Orthopedic Surgery Excellence Award(tm) from HealthGrades,
the nation's leading health care ratings company.
 
Visit Somerset Medical Center's Web site - www.somersetmedicalcenter.com -
for news, event listings, health information and more. And, find us on Facebook
and "Become a Fan" for up-to-the-minute medical center news.
From billodonnell <@t> catholichealth.net  Fri May 29 09:38:38 2009
From: billodonnell <@t> catholichealth.net (O'Donnell, Bill)
Date: Fri May 29 09:38:50 2009
Subject: [Histonet] Multi-specimen tissue micro array on the cheap
In-Reply-To: 
References: 
	
Message-ID: 

To all who responded to my last post concerning an inexpensive method to make micro arrays: 

I will be more than happy to share, once I actually get around to writing it up. Now I have the impetus to do so.

I'll get it done in the next week and will post in general to this list-serve.

William (Bill) O'Donnell, HT (ASCP) QIHC 
Lead Histologist
Good Samaritan Hospital
10 East 31st Street
Kearney, NE 68847 




-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Andrea Grantham
Sent: Thursday, May 28, 2009 5:08 PM
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Advice on fatty liver sectioning

-18? is the temp I use when I section liver for ORO and I usually cut the sections at 5 microns. It sounds like your tissue was not frozen correctly.

Andi



Andrea Grantham, HT (ASCP)
Senior Research Specialist
University of Arizona
Cell Biology and Anatomy
Histology Service Laboratory
P.O.Box 245044
Tucson, AZ 85724

algranth@email.arizona.edu
Tel: 520.626.4415     Fax: 520.626.2097

"happy slicing and dicing and may all your stains work perfectly" - Paula Sicurello P Please consider the environment before printing this email.



On May 28, 2009, at 12:20 PM, Kinsley, David wrote:

> Hi,
> I have some very fatty mouse livers that I have to obtain frozen 
> sections for Oil red O and IHC.  I have tried sectioning from -18 to 
> -30 degrees and from 5um to 10um and the liver just seems to compress, 
> pull away from the OCT, or roll up on itself while the OCT sections 
> well.  When I section below -30 parts of the liver have chatter in 
> them as if they are too cold.  Does anyone have any suggestions on 
> what the best cutting temperature would be for these?
>
> I have successfully cut normal and less fatty livers at 5um ad 10 um 
> at -18 and -25 degrees respectively, it is these extremely fatty 
> livers that are giving me problems.  Any advice is appreciated.
>
> Dave
>
>
>
>
>
> *********************************************************************
> This message and any attachments are solely for the intended 
> recipient. If you are not the intended recipient, disclosure, copying, 
> use or distribution of the information included in this message is 
> prohibited -- Please immediately and permanently delete.
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>

_______________________________________________
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From talulahgosh <@t> gmail.com  Fri May 29 09:41:57 2009
From: talulahgosh <@t> gmail.com (Emily Sours)
Date: Fri May 29 09:42:05 2009
Subject: [Histonet] Microtome Problems
In-Reply-To: 
References: 
	
Message-ID: 

The microtome jumping forward sounds like something isn't locking correctly
anymore--either the knife stage or the section thickness dial.
When our microtomes do that, I just try jiggling every part separately to
make sure everything is locked.

Emily

"People say: 'How you can see hummingbirds, roses, and orchids and not
believe in the Lord's splendour?.'..Imagine an African boy with a parasitic
worm boring into his eye. If you tell me God not only created but cares for
us all, what about that boy? Are you telling me he says: 'I understand. God
deliberately created a worm that's going to blind me?' I find that
intolerable."
--David Attenborough


On Fri, May 29, 2009 at 10:38 AM, Rathborne, Toni <
trathborne@somerset-healthcare.com> wrote:

> Usually the service techs that come in will give a 30 day call-back period
> that is included as part of the service call. I would have them come back
> until it is working properly.
>
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of
> Veterinary Services Laboratory
> Sent: Friday, May 29, 2009 10:12 AM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Microtome Problems
>
>
>
>
> Hello All
>
>
>
> I'm new to the group and histo. (3 years) so far I'm getting a lot of
> useful
> tips from all the messages.  Thank you.
>
>
>
> I have two microtomes that were working almost perfectly before I had a
> technician service them.  Now, the Leica 2035 Jung biocuts' blade holder is
> twisting to the side (going up at one end).  I have tried tightening,
> slacking, and cleaning the entire assembly to no avail.  Does anyone know
> of
> where I might be able to source one or even how to fix the one I have.
>
>
>
> Secondly, the Leitz 1512 rotary microtome, is now jumping forward and
> giving
> a loud clanking noise.  It takes huge chunks out of the tissue.
>
>
>
> Any and all help is appreciated,
>
>
>
> Thank you
>
> Sharon
>
>
>
> Ms. Sharon Drayton
>
> Veterinary Services Laboratory
>
> Ministry of Agriculture & Rural Development
>
> The Pine
>
> St. Michael BB11091
>
> Barbados
>
>
>
> Tel:  (246) 427-5492 or (246) 427-5073
>
> Fax: (246)426-7517
>
> Email:  vetlab@caribsurf.com
>
> Website:  www.agriculture.gov.bb
>
>
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> CONFIDENTIALITY NOTICE
> This message and any included attachments are from
> Somerset Medical Center and are intended only for the addressee.
> The information contained in this message is confidential
> and may contain privileged, confidential, proprietary and/or
> trade secret information entitled to protection and/or exemption
> from disclosure under applicable law. Unauthorized forwarding,
> printing, copying, distribution, or use of such information is
> strictly prohibited and may be unlawful. If you are not the
> addressee, please promptly delete this message and notify
> the sender of the delivery error by e-mail or you may call
> Somerset Medical Center's computer Help Desk at
> 908-685-2200, ext. 4050.
> --------------------------------------------------------------
> Somerset Medical Center is the recipient of the
> 2009 Orthopedic Surgery Excellence Award(tm) from HealthGrades,
> the nation's leading health care ratings company.
>
> Visit Somerset Medical Center's Web site - www.somersetmedicalcenter.com -
> for news, event listings, health information and more. And, find us on
> Facebook
> and "Become a Fan" for up-to-the-minute medical center news.
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
From jmcgough <@t> clinlab.com  Fri May 29 09:56:51 2009
From: jmcgough <@t> clinlab.com (Jason McGough)
Date: Fri May 29 09:56:56 2009
Subject: [Histonet] Sakura film coverslipper xylene residue issue
In-Reply-To: 
References: 
Message-ID: <20090529085651.6pa1aldhvb44sw0s@mail.clinlab.com>

It has nothing to do with your coverslipper. Ours is from the mid 90's  
and we have had the brown spots a couple of times. We found that there  
to be water in our xylene. The little water droplets on the side of a  
container will cause this. Make sure everything that holds xylene is  
completely dry. Once we did this, we had no more brown spots. Good luck!

As far as xylene residue, it is your dispensing volume on the  
coverslipper that is causing this. Try turning the flow down and see  
what happens.

Jason McGough HT(ASCP)
Account Representative-Anatomic Pathology
Clinical Laboratory of the Black Hills
2805 5th Street Suite 210
Rapid City, SD 57701
605-343-2267
jmcgough@clinlab.com


Quoting Jackie M O'Connor :

> I have an older Sakura coverslipper, circa 1997ish.  We have noticed an
> increasing number of brown spots on tissue sections recently.   This does
> not occur if the same slides are covered with glass coverslips.   It does
> not look like a drying artefact, occurs randomly throughout tissue types,
> location on tissue, i.e., edge, center.   I've even showed these spots to
> the rep who admitted she has never seen this artefact before.   We use
> only the Tissue Tek tape, we maintain the equipment on a regular basis.
> The xylene flow is good.   Our only problem are these random brown spots.
>  Any suggestions?
> Jackie O'
>
>
>
> "Karin Groeger" 
> Sent by: histonet-bounces@lists.utsouthwestern.edu
> 05/29/2009 08:19 AM
>
> To
> "Scott" , 
> cc
>
> Subject
> RE: [Histonet] Sakura film coverslipper xylene residue issue
>
>
>
>
>
>
> Hi Scott,
>   We place ours in front of a fan to dry if we are in a hurry, even if
> we air dry them we do not have a problem with the xylene residue,
> perhaps you have  a bad batch, check your lot # and call Sakura.  We
> coverslip over 1,000 slides a day and have not seen this.
>
> Karin Groeger
>
> Histology Supervisor
>
> US LABS, Irvine,CA
>
> 949-450-0145 ext. 649
>
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Scott
> Sent: Thursday, May 28, 2009 10:03 PM
> To: Histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Sakura film coverslipper xylene residue issue
>
> Hi,
>     We have a newer Sakura film coverslipper, does anyone have
> experience with this piece of equipment. I have a problem with the
> slides having a light xylene residue, after the slides come off of the
> coverslipper.
>
> How do you dry them? If I leave the slides on the coverslipper to air
> dry the xylene residue is noticeable. If I lightly wipe off all of the
> slides as they come off the coverslipper they look clean.
>
> How do you guys do it? I think it is a waste of time to wipe off all of
> the slides. I don't think the xylene flow is too high.
>
> Thanks for your help,
>
> Scott Hendricksen HT(ASCP)
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> Confidentiality Notice: This message, including any attachments, may
> contain CONFIDENTIAL AND/OR LEGALLY PRIVILEGED information. The
> information is intended for use by the individual named above and may not
> be disseminated to any other party without US LABS' written permission. If
> you are not the intended recipient, or the employee or agent responsible
> for delivering this information to the intended recipient, you are hereby
> notified that any dissemination, disclosure, distribution, copying or
> taking of any action in reliance on the contents of this information is
> strictly prohibited. If you have received this information in error,
> please notify US LABS immediately at 1-888-450-0145 attn: Compliance
> Department to arrange for return of this message including all
> attachments.
>
> _______________________________________________
> Histonet mailing list
> Histonet@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 azhistotech <@t> cox.net  Fri May 29 10:48:16 2009
From: azhistotech <@t> cox.net (azhistotech@cox.net)
Date: Fri May 29 10:48:20 2009
Subject: [Histonet] Looking for travel job in Ca or Florida
Message-ID: <20090529114816.2H1BE.137513.imail@fed1rmwml44>

Hi Netters,
Is anyone looking for a Histologist in California or Florida? I am an HT, (ascp) with Florida license, 18 years experience, 5 years management and know all aspects of Histology. Looking for a Travel position or short term assignment. Please respond to this email and I can forward you my resume. Thank you..

From Robert.Lott <@t> trinitymedicalonline.com  Fri May 29 11:35:16 2009
From: Robert.Lott <@t> trinitymedicalonline.com (Lott, Robert)
Date: Fri May 29 11:35:56 2009
Subject: [Histonet] MAP-2 antibody
Message-ID: <4A3619571D9F6C4CB79C980E91DBE4E60106801E@TNTRIEXEVS03.triadhospitals.net>

Hi Everyone,

One of our pathologists is interested in MAP-2 IHC for the following
application:

 

Microtubule Associated Protein-2 Is a Sensitive Marker of Primary and
Metastatic Neuroblastoma.                         

Krishnan C, Heerema-McKenney A, Arber DA.  Department of Pathology,
Stanford University, Stanford, CA.

 

Comment: Microtubule associated protein-2 (MAP-2) is a protein expressed
in high levels in cells derived from the neural crest.  

It is a very robust marker of neuronal differentiation and has been used
most often in evaluating neuronal differentiation in tumors 

of the central nervous system.  MAP-2 showed comparable sensitivity in
staining primary neuroblastomas 

(pre- and post-treatment samples) as compared to synaptophysin,
chromogranin, CD56, and beta-catenin.  

In contrast to other markers of neuroblastoma, MAP-2 did not show
significant cross reactivity to native bone marrow 

precursors, thus eliminating a potential source of confusion.

 

Does anyone use this antibody for this or like applications....

I have looked and am "fairly" sure that there is not an IVD available,
but would still be interested in knowing which antibody you use.

 

Thanks,

 

Robert L. Lott, HTL(ASCP) / Manager, Anatomic Pathology 

Trinity Medical Center/ LabFirst/ 800 Montclair Road / Birmingham, AL
35213/ 205-592-5387 

 



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From mari.ann.mailhiot <@t> leica-microsystems.com  Fri May 29 11:50:19 2009
From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com)
Date: Fri May 29 11:50:34 2009
Subject: [Histonet] Microtome Problems
In-Reply-To: 
Message-ID: 

HI Sharon

I am going to refer your email to my collegues in Germany. They will be
able to get you the help you need and all the information..

Best Regards

Mari Ann Mailhiot BA HT ASCP
Application Specialist/Trainer
Leica Microsystems
Biosystems Division
Technical Assistance Center
800 248 0123 x7267
847 236 3063 fax
mari.ann.mailhiot@leica-microsystems.com
www.leica-microsystems.com


                                                                           
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             histonet-bounces@                                     Subject 
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             ern.edu                                                       
                                                                           
                                                                           
             05/29/2009 09:11                                              
             AM                                                            
                                                                           





Hello All



I'm new to the group and histo. (3 years) so far I'm getting a lot of
useful
tips from all the messages.  Thank you.



I have two microtomes that were working almost perfectly before I had a
technician service them.  Now, the Leica 2035 Jung biocuts' blade holder is
twisting to the side (going up at one end).  I have tried tightening,
slacking, and cleaning the entire assembly to no avail.  Does anyone know
of
where I might be able to source one or even how to fix the one I have.



Secondly, the Leitz 1512 rotary microtome, is now jumping forward and
giving
a loud clanking noise.  It takes huge chunks out of the tissue.



Any and all help is appreciated,



Thank you

Sharon



Ms. Sharon Drayton

Veterinary Services Laboratory

Ministry of Agriculture & Rural Development

The Pine

St. Michael BB11091

Barbados



Tel:  (246) 427-5492 or (246) 427-5073

Fax: (246)426-7517

Email:  vetlab@caribsurf.com

Website:  www.agriculture.gov.bb





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From juditw <@t> u.washington.edu  Fri May 29 12:00:26 2009
From: juditw <@t> u.washington.edu (Judith L. Williams)
Date: Fri May 29 12:00:39 2009
Subject: [Histonet] Horseshoe crab..help!
In-Reply-To: 
Message-ID: 

Hi all- chitin (invertebrate shell) is not the same as keratin (fingernails)but that may work! In the past I have found with the inverts I did histo on that long term storage in 70% ETOH rendered the chitin very very soft. so - maybe try this. Another thing to try is chitinase.
good luck with the invert project!
Judy




On Thu, 28 May 2009, Ingles Claire  wrote:

> Anyone please correct me if I'm wrong, but is chitin similar to fingernails? If so, we just use Nair for a bit. You may need to use it for a few hours since the shell in horseshoe crabs is harder than regular fingernails.
> Claire
>
> ________________________________
>
> From: histonet-bounces@lists.utsouthwestern.edu on behalf of Deborah Faichney
> Sent: Thu 5/28/2009 4:56 AM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Horseshoe crab..help!
>
>
>
> Hi all,
>
> I have a Masters student here who has Horseshoe crab tissues fixed in 10% NBF.  Most of the tissues have been sectioned successfully but the eyes are surrounded by armour plated chitin! (she broke two Dremel drills trying to cut the carapace and finally tin snips had to be used to cut them out!!).
> Any thoughts on how to soften them?  Looking around on the Net, I have found  that processing and clearing through Chloroform may soften, also a solution called Diaphanol has been recommended.
> At the moment they have been two days in a 1 part acetic acid to 5 parts NBF with no effect.
> We only have a couple of weeks to do this work so cannot adopt any trial or lengthy methods.
>
> Is this possible?  I like a challenge, but suspect this needs a miracle.  Its no wonder these beasties have been on the planet for so long.
> Many thanks
>
> Debbie Faichney
> Histopathology
> Institute of Aquaculture
> University of Stirling
> Stirling, FK7 7QS
> Scotland
> Uk
>
>
>
>
> --
> Academic Excellence at the Heart of Scotland.
> The University of Stirling is a charity registered in Scotland,
> number SC 011159.
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>


Judith Williams, PhD, HT(ASCP)
Research Scientist
Department of Comparative Medicine
University of Washington
Seattle, WA 98195



From Simoskevitz <@t> Osteotech.com  Fri May 29 12:58:39 2009
From: Simoskevitz <@t> Osteotech.com (Simoskevitz@Osteotech.com)
Date: Fri May 29 12:58:01 2009
Subject: [Histonet] Trichrome stain in GMA
In-Reply-To: <20090529170152.32E3E31A75D@l3inpf1.contentcatcher.com>
Message-ID: 

Does anyone have a good protocol for a Trichrome stain in glycol
methacrylate bone sections.  I keep getting non specific staining for
demineralized bone sections.  I am using a microwave technique that stains
my top section one way and my bottom section another.  I have cut at
different thicknesses tried different times and rinsing techniques.
Sometimes half of my section will stain one way while the other have stains
different.  Any help would be greatly appreciated.

Ricki

*********************************************
Ricki Simoskevitz, Research Associate III
Osteotech Inc.
51 James Way
Eatontown, NJ 07724
Phone: (732) 542-2800 X6328
Fax: (732) 935-1298
**********************************************


From melissa.mazan <@t> tufts.edu  Fri May 29 13:18:31 2009
From: melissa.mazan <@t> tufts.edu (Melissa Mazan)
Date: Fri May 29 13:18:37 2009
Subject: [Histonet] CD11c for mouse tissue
In-Reply-To: <200905291702.n4TH2kUZ003928@mail-proofpoint-2a>
References: <200905291702.n4TH2kUZ003928@mail-proofpoint-2a>
Message-ID: <4A2026F7.3000204@tufts.edu>

Hi all - I'm looking for a good antibody to use on mouse tissue (frozen) 
to look for CD11c - does anyone have a suggestion for a good antibody 
source? Thanks in advance - Melissa

histonet-request@lists.utsouthwestern.edu wrote:
> Send Histonet mailing list submissions to
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>
> Today's Topics:
>
>    1. RE: Sakura film coverslipper xylene residue issue (Jason McGough)
>    2. Looking for travel job in Ca or Florida (azhistotech@cox.net)
>    3. MAP-2 antibody (Lott, Robert)
>    4. Re: Microtome Problems (mari.ann.mailhiot@leica-microsystems.com)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Fri, 29 May 2009 08:56:51 -0600
> From: Jason McGough 
> Subject: RE: [Histonet] Sakura film coverslipper xylene residue issue
> To: Jackie M O'Connor 
> Cc: Histonet@lists.utsouthwestern.edu, Karin Groeger
> 	,	histonet-bounces@lists.utsouthwestern.edu
> Message-ID: <20090529085651.6pa1aldhvb44sw0s@mail.clinlab.com>
> Content-Type: text/plain;	charset=ISO-8859-1;	DelSp="Yes";
> 	format="flowed"
>
> It has nothing to do with your coverslipper. Ours is from the mid 90's  
> and we have had the brown spots a couple of times. We found that there  
> to be water in our xylene. The little water droplets on the side of a  
> container will cause this. Make sure everything that holds xylene is  
> completely dry. Once we did this, we had no more brown spots. Good luck!
>
> As far as xylene residue, it is your dispensing volume on the  
> coverslipper that is causing this. Try turning the flow down and see  
> what happens.
>
> Jason McGough HT(ASCP)
> Account Representative-Anatomic Pathology
> Clinical Laboratory of the Black Hills
> 2805 5th Street Suite 210
> Rapid City, SD 57701
> 605-343-2267
> jmcgough@clinlab.com
>
>
> Quoting Jackie M O'Connor :
>
>   
>> I have an older Sakura coverslipper, circa 1997ish.  We have noticed an
>> increasing number of brown spots on tissue sections recently.   This does
>> not occur if the same slides are covered with glass coverslips.   It does
>> not look like a drying artefact, occurs randomly throughout tissue types,
>> location on tissue, i.e., edge, center.   I've even showed these spots to
>> the rep who admitted she has never seen this artefact before.   We use
>> only the Tissue Tek tape, we maintain the equipment on a regular basis.
>> The xylene flow is good.   Our only problem are these random brown spots.
>>  Any suggestions?
>> Jackie O'
>>
>>
>>
>> "Karin Groeger" 
>> Sent by: histonet-bounces@lists.utsouthwestern.edu
>> 05/29/2009 08:19 AM
>>
>> To
>> "Scott" , 
>> cc
>>
>> Subject
>> RE: [Histonet] Sakura film coverslipper xylene residue issue
>>
>>
>>
>>
>>
>>
>> Hi Scott,
>>   We place ours in front of a fan to dry if we are in a hurry, even if
>> we air dry them we do not have a problem with the xylene residue,
>> perhaps you have  a bad batch, check your lot # and call Sakura.  We
>> coverslip over 1,000 slides a day and have not seen this.
>>
>> Karin Groeger
>>
>> Histology Supervisor
>>
>> US LABS, Irvine,CA
>>
>> 949-450-0145 ext. 649
>>
>> -----Original Message-----
>> From: histonet-bounces@lists.utsouthwestern.edu
>> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Scott
>> Sent: Thursday, May 28, 2009 10:03 PM
>> To: Histonet@lists.utsouthwestern.edu
>> Subject: [Histonet] Sakura film coverslipper xylene residue issue
>>
>> Hi,
>>     We have a newer Sakura film coverslipper, does anyone have
>> experience with this piece of equipment. I have a problem with the
>> slides having a light xylene residue, after the slides come off of the
>> coverslipper.
>>
>> How do you dry them? If I leave the slides on the coverslipper to air
>> dry the xylene residue is noticeable. If I lightly wipe off all of the
>> slides as they come off the coverslipper they look clean.
>>
>> How do you guys do it? I think it is a waste of time to wipe off all of
>> the slides. I don't think the xylene flow is too high.
>>
>> Thanks for your help,
>>
>> Scott Hendricksen HT(ASCP)
>> _______________________________________________
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>
>>
>> Confidentiality Notice: This message, including any attachments, may
>> contain CONFIDENTIAL AND/OR LEGALLY PRIVILEGED information. The
>> information is intended for use by the individual named above and may not
>> be disseminated to any other party without US LABS' written permission. If
>> you are not the intended recipient, or the employee or agent responsible
>> for delivering this information to the intended recipient, you are hereby
>> notified that any dissemination, disclosure, distribution, copying or
>> taking of any action in reliance on the contents of this information is
>> strictly prohibited. If you have received this information in error,
>> please notify US LABS immediately at 1-888-450-0145 attn: Compliance
>> Department to arrange for return of this message including all
>> attachments.
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>     
>
>
>
>
>
> ------------------------------
>
> Message: 2
> Date: Fri, 29 May 2009 8:48:16 -0700
> From: 
> Subject: [Histonet] Looking for travel job in Ca or Florida
> To: histonet@lists.utsouthwestern.edu
> Message-ID: <20090529114816.2H1BE.137513.imail@fed1rmwml44>
> Content-Type: text/plain; charset=utf-8
>
> Hi Netters,
> Is anyone looking for a Histologist in California or Florida? I am an HT, (ascp) with Florida license, 18 years experience, 5 years management and know all aspects of Histology. Looking for a Travel position or short term assignment. Please respond to this email and I can forward you my resume. Thank you..
>
>
>
> ------------------------------
>
> Message: 3
> Date: Fri, 29 May 2009 11:35:16 -0500
> From: "Lott, Robert" 
> Subject: [Histonet] MAP-2 antibody
> To: 
> Cc: Christa Hladik 
> Message-ID:
> 	<4A3619571D9F6C4CB79C980E91DBE4E60106801E@TNTRIEXEVS03.triadhospitals.net>
> 	
> Content-Type: text/plain;	charset="us-ascii"
>
> Hi Everyone,
>
> One of our pathologists is interested in MAP-2 IHC for the following
> application:
>
>  
>
> Microtubule Associated Protein-2 Is a Sensitive Marker of Primary and
> Metastatic Neuroblastoma.                         
>
> Krishnan C, Heerema-McKenney A, Arber DA.  Department of Pathology,
> Stanford University, Stanford, CA.
>
>  
>
> Comment: Microtubule associated protein-2 (MAP-2) is a protein expressed
> in high levels in cells derived from the neural crest.  
>
> It is a very robust marker of neuronal differentiation and has been used
> most often in evaluating neuronal differentiation in tumors 
>
> of the central nervous system.  MAP-2 showed comparable sensitivity in
> staining primary neuroblastomas 
>
> (pre- and post-treatment samples) as compared to synaptophysin,
> chromogranin, CD56, and beta-catenin.  
>
> In contrast to other markers of neuroblastoma, MAP-2 did not show
> significant cross reactivity to native bone marrow 
>
> precursors, thus eliminating a potential source of confusion.
>
>  
>
> Does anyone use this antibody for this or like applications....
>
> I have looked and am "fairly" sure that there is not an IVD available,
> but would still be interested in knowing which antibody you use.
>
>  
>
> Thanks,
>
>  
>
> Robert L. Lott, HTL(ASCP) / Manager, Anatomic Pathology 
>
> Trinity Medical Center/ LabFirst/ 800 Montclair Road / Birmingham, AL
> 35213/ 205-592-5387 
>
>  
>
>
>
> --------------------------------------------------------------------------
> Disclaimer: This electronic message may contain information that is
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>
> ------------------------------
>
> Message: 4
> Date: Fri, 29 May 2009 11:50:19 -0500
> From: mari.ann.mailhiot@leica-microsystems.com
> Subject: Re: [Histonet] Microtome Problems
> To: "Veterinary Services Laboratory" 
> Cc: histonet@lists.utsouthwestern.edu
> Message-ID:
> 	
> 	
> Content-Type: text/plain; charset=US-ASCII
>
> HI Sharon
>
> I am going to refer your email to my collegues in Germany. They will be
> able to get you the help you need and all the information..
>
> Best Regards
>
> Mari Ann Mailhiot BA HT ASCP
> Application Specialist/Trainer
> Leica Microsystems
> Biosystems Division
> Technical Assistance Center
> 800 248 0123 x7267
> 847 236 3063 fax
> mari.ann.mailhiot@leica-microsystems.com
> www.leica-microsystems.com
>
>
>                                                                            
>              "Veterinary                                                   
>              Services                                                      
>              Laboratory"                                                To 
>               
>              .com>                                                      cc 
>              Sent by:                                                      
>              histonet-bounces@                                     Subject 
>              lists.utsouthwest         [Histonet] Microtome Problems       
>              ern.edu                                                       
>                                                                            
>                                                                            
>              05/29/2009 09:11                                              
>              AM                                                            
>                                                                            
>
>
>
>
>
> Hello All
>
>
>
> I'm new to the group and histo. (3 years) so far I'm getting a lot of
> useful
> tips from all the messages.  Thank you.
>
>
>
> I have two microtomes that were working almost perfectly before I had a
> technician service them.  Now, the Leica 2035 Jung biocuts' blade holder is
> twisting to the side (going up at one end).  I have tried tightening,
> slacking, and cleaning the entire assembly to no avail.  Does anyone know
> of
> where I might be able to source one or even how to fix the one I have.
>
>
>
> Secondly, the Leitz 1512 rotary microtome, is now jumping forward and
> giving
> a loud clanking noise.  It takes huge chunks out of the tissue.
>
>
>
> Any and all help is appreciated,
>
>
>
> Thank you
>
> Sharon
>
>
>
> Ms. Sharon Drayton
>
> Veterinary Services Laboratory
>
> Ministry of Agriculture & Rural Development
>
> The Pine
>
> St. Michael BB11091
>
> Barbados
>
>
>
> Tel:  (246) 427-5492 or (246) 427-5073
>
> Fax: (246)426-7517
>
> Email:  vetlab@caribsurf.com
>
> Website:  www.agriculture.gov.bb
>
>
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ______________________________________________________________________
> This email has been scanned by the MessageLabs Email Security System.
> For more information please visit http://www.messagelabs.com/email 
> ______________________________________________________________________
>
>
>
> ------------------------------
>
> _______________________________________________
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>
> End of Histonet Digest, Vol 66, Issue 35
> ****************************************
>   

-- 
Melissa R. Mazan, DVM, Diplomate ACVIM
Associate Professor and Director of Equine Sports Medicine
Department of Clinical Sciences
Tufts Cummings School of Veterinary Medicine
200 Westborough Road
North Grafton,MA 01536
tel:	508-839-5395
fax:	508-839-7903
email:	melissa.mazan@tufts.edu


From contact <@t> excaliburpathology.com  Fri May 29 13:43:02 2009
From: contact <@t> excaliburpathology.com (Paula Pierce)
Date: Fri May 29 13:43:07 2009
Subject: [Histonet] Trichrome stain in GMA
In-Reply-To: 
References: 
Message-ID: <744004.40779.qm@web1111.biz.mail.sk1.yahoo.com>

Hello,

you are getting darker staining on the top sections because the solution is hotter at the top. You should stir the solution after heating to distribute the heat. I use a plastic transfer pipet to do this. A few draws and empties will stir the solution well.

Paula




________________________________
From: "Simoskevitz@Osteotech.com" 
To: histonet@lists.utsouthwestern.edu
Sent: Friday, May 29, 2009 12:58:39 PM
Subject: [Histonet] Trichrome stain in GMA

Does anyone have a good protocol for a Trichrome stain in glycol
methacrylate bone sections.? I keep getting non specific staining for
demineralized bone sections.? I am using a microwave technique that stains
my top section one way and my bottom section another.? I have cut at
different thicknesses tried different times and rinsing techniques.
Sometimes half of my section will stain one way while the other have stains
different.? Any help would be greatly appreciated.

Ricki

*********************************************
Ricki Simoskevitz, Research Associate III
Osteotech Inc.
51 James Way
Eatontown, NJ 07724
Phone: (732) 542-2800 X6328
Fax: (732) 935-1298
**********************************************


_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
From angrodoronar <@t> gmail.com  Fri May 29 14:22:39 2009
From: angrodoronar <@t> gmail.com (Mark H)
Date: Fri May 29 14:22:43 2009
Subject: [Histonet] Still looking for employment in Phoenix or Tucson Arizona
Message-ID: <16c76c910905291222t5f881d58p7f2ae7ad693d5834@mail.gmail.com>

Just wondering anyone knows of any new openings.

I?m a recent Arizona Pima Community College graduate in histology
seeking full time employment in either the Tucson or Phoenix area.
Current member of ASH and NSH, as well as 1 year experience in co-ops
at Ventana Medical Systems and North West Medical Center in Tucson.

Mark

520-904-8005 or respond to this E-mail

Thank You

From Montina.VanMeter <@t> pbrc.edu  Fri May 29 15:07:37 2009
From: Montina.VanMeter <@t> pbrc.edu (Montina Van Meter)
Date: Fri May 29 15:07:43 2009
Subject: [Histonet] 2009 Louisiana Society for Histotechnology
	Symposium/Convention Invitation
Message-ID: <4FE7FB862E90E448AE32388E759220E50140B2C2@pbrcas31.pbrc.edu>

Fellow Histonetters,

     The Louisiana Society for Histotechnology is pleased to announce
the 26th Annual Symposium/Convention: 
                                     
                                                              "Your
Histeaux Surplus Package"

 
June 12 & 13, 2009

 
at the

 
Bourbon Orleans Hotel

 
717 Orleans St.
                  
 
New Orleans, LA  70117

 
www.bourbonorleans.com



 1.  Walk-ins are always welcome!  If you are planning on registering
the day of the meeting we would appreciate it if you 
      would send us an email or call Dixie or Tina prior to the meeting,
which will allow the LSH to make arrangements for 
      workshop handouts and food reservations.  Pre-registrants will
automatically receive a complimentary buffet lunch each 
      day!  Please make additional copies of your registration form for
co-workers in your lab or facility that might be interested   
      in attending.  The LSH would also like to extend the invitation to
our fellow technologists and pathologists from 
      surrounding states.  We encourage everyone to attend in order to
build our networking potential, earn those valuable 
      CEU's and enjoy the beautiful city of New Orleans.  

 2.  The LSH room rates will be honored as long as rooms are available.
For those attendees who may want to visit the 
      beautiful sights and sounds of New Orleans, the Bourbon Orleans
Hotel will honor the room rates for three days prior and 
      three days after the meeting.  Special on-site parking rates for
LSH attendees is available as well as public parking 
      around Jackson Square.  For reservations call 1-504-523-2222 or
1-866-513-9744 and mention you are with the LSH 
      group.  Do to relocation of many of our members we would ask
everyone who would like to receive a 
      brochure/membership form via email or fax, to please contact Tina
Van Meter at 225-603-0953, 
      vanmetmj@pbrc.edu or Dixie Benoit at 337-233-1951,
dixiehistochick@bellsouth.net .  

 
3.   There are many local attractions going on during the week of our
symposium such as: The Cajun - Zydeco Festival,  
      Seafood Festival and the Creole Tomato Festival all in the French
Quarter/Jackson Square area.  Free admission to the 
      festivals.   Check out the links below:                         
                                                                 
 
http://www.neworleansonline.com/news/2009/May/vieuxtodo.html

 
http://ww.neworleansonline.com/neworleans/tours/


4.   Employment Postings:  The LSH will have a "Job Board" for anyone
who would like to advertise opportunities for  
       employment at their facility.  Please contact us via email, phone
or at the meeting. 


     
 
5.   2009 LSH State Meeting Workshops:

     WS#1 - Am I Really Ready for this CAP Inspection?

     WS#2 - Mouse to Horse: It's Not Human

     WS#3 - Breast Cancer and the Standardization of HER2/neu Testing

     WS#4 - Contemporary Trends in Immunohistochemistry 

     WS#5 - Troubleshooting Routine Special Stains 

     WS#6 - Which Code Do I Use?  (CPT Coding)

     WS#7 - Are You REALLY Ready for the Next Catastrophic Event That
Will Affect Your Hospital or City?

     WS#8 - 2 Crazy Cat Ladies Give Their Opinions of Life, Liberty, and
Lab Management    

 We have a variety of topics presented by experienced speakers that
promises to benefit everyone.  The attendees will have  
 access to several scientific vendor exhibits during the entire
symposium.  The LSH would love to have y'all come on down to 
 the Bourbon Orleans Hotel in the French Quarter!

 

Hope to see you in June!

The LSH Committee



Montina J. Van Meter, HT (ASCP)
Lab Manager
Autonomic Neuroscience
Pennington Biomedical Research Center
6400 Perkins Rd.
Baton Rouge, LA  70791
225-763-2564



From gayle.callis <@t> bresnan.net  Sat May 30 09:45:09 2009
From: gayle.callis <@t> bresnan.net (gayle callis)
Date: Sat May 30 09:45:27 2009
Subject: [Histonet] RE: CD11c for murine tissu
Message-ID: <000501c9e135$3c559dd0$b500d970$@callis@bresnan.net>

You wrote:  Hi all - I'm looking for a good antibody to use on mouse tissue
(frozen) 

to look for CD11c - does anyone have a suggestion for a good antibody 

source? Thanks in advance - Melissa

 

We us CD11c, HL3 clone, Armenian Hamster anti Mouse.  Negative control is
Armenian Hamster IgG1 from BD Pharmingen/Invitrogen.  Tissues are fresh
frozen sections fixed with our favorite acetone/absolute ethanol fixative,
although you can use 4C acetone for 10 min on overnight air dried sections.
I will be happy to send the other fixative recipe if you want it. 

 

Gayle M. Callis

HTL(ASCP)HT,MT

Bozeman MT 

 

 

 

From gayle.callis <@t> bresnan.net  Sat May 30 10:00:44 2009
From: gayle.callis <@t> bresnan.net (gayle callis)
Date: Sat May 30 10:01:04 2009
Subject: [Histonet] RE: trichrome stain on GMA bone sections
Message-ID: <000f01c9e137$6ba0c900$42e25b00$@callis@bresnan.net>

Ricki, 

 

I have a Modified Oserhoff (sp) Massons Trichrome method for bone sections
in GMA but it requires a really thin sections (1 um) and no microwave
heating.  Bob Skinner had great success with this modified staining method.
There  is also a publication in J of Histotechnology on this protocol, many
years back.    I can dig  the method  out of my pre-computer day files for
you if you wish.  

 

Doing trichromes on GMA embedded tissues is tricky due to the nature of dyes
(aniline blue) and the dye molecular weights.  Often the GMA stains blue
too??  

 

Gayle M. Callis 

HTL(ASCP)HT,MT

Bozeman MT 

From robert_schoonhoven <@t> yahoo.com  Sat May 30 11:37:26 2009
From: robert_schoonhoven <@t> yahoo.com (Robert Schoonhoven)
Date: Sat May 30 11:37:29 2009
Subject: [Histonet] Looking for new and used equipment
Message-ID: <485074.32572.qm@web31101.mail.mud.yahoo.com>

I am working as a consultant to a new (from scratch) histology lab in the Kentucky - Indiana area and we are looking at both new (some), and refurbished (mostly) equipment.  I would appreciate an off list reply from suppliers.

Thanks,
Bob


 Robert Schoonhoven, HT/HTL (ASCP)


robert_schoonhoven@yahoo.com


      
From pruegg <@t> ihctech.net  Sat May 30 11:42:41 2009
From: pruegg <@t> ihctech.net (Patsy Ruegg)
Date: Sat May 30 11:42:46 2009
Subject: [Histonet] RE: trichrome stain on GMA bone sections
In-Reply-To: <000f01c9e137$6ba0c900$42e25b00$@callis@bresnan.net>
References: <000f01c9e137$6ba0c900$42e25b00$@callis@bresnan.net>
Message-ID: <73C8AA35EBAC4C9DADC9EEC73B92CBE4@prueggihctechlt>

I have a trichrome for GMA as well, it uses green instead of aniline blue
and some other modifications.
Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net 
www.ihcrg.org

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of gayle callis
Sent: Saturday, May 30, 2009 9:01 AM
To: 'Histonet'
Subject: [Histonet] RE: trichrome stain on GMA bone sections

Ricki, 

 

I have a Modified Oserhoff (sp) Massons Trichrome method for bone sections
in GMA but it requires a really thin sections (1 um) and no microwave
heating.  Bob Skinner had great success with this modified staining method.
There  is also a publication in J of Histotechnology on this protocol, many
years back.    I can dig  the method  out of my pre-computer day files for
you if you wish.  

 

Doing trichromes on GMA embedded tissues is tricky due to the nature of dyes
(aniline blue) and the dye molecular weights.  Often the GMA stains blue
too??  

 

Gayle M. Callis 

HTL(ASCP)HT,MT

Bozeman MT 

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From pruegg <@t> ihctech.net  Sat May 30 11:44:14 2009
From: pruegg <@t> ihctech.net (Patsy Ruegg)
Date: Sat May 30 11:44:18 2009
Subject: [Histonet] RE: CD11c for murine tissu
In-Reply-To: <000501c9e135$3c559dd0$b500d970$@callis@bresnan.net>
References: <000501c9e135$3c559dd0$b500d970$@callis@bresnan.net>
Message-ID: <21D20198BF6341BA96FB25ABD32ECF65@prueggihctechlt>

An alternative to cd11c for macrophages is rat anti mouse f4/80 which I use
from Serotec, it works in frozen and ffpe tissue.
Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net 
www.ihcrg.org


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of gayle callis
Sent: Saturday, May 30, 2009 8:45 AM
To: 'Histonet'
Subject: [Histonet] RE: CD11c for murine tissu

You wrote:  Hi all - I'm looking for a good antibody to use on mouse tissue
(frozen) 

to look for CD11c - does anyone have a suggestion for a good antibody 

source? Thanks in advance - Melissa

 

We us CD11c, HL3 clone, Armenian Hamster anti Mouse.  Negative control is
Armenian Hamster IgG1 from BD Pharmingen/Invitrogen.  Tissues are fresh
frozen sections fixed with our favorite acetone/absolute ethanol fixative,
although you can use 4C acetone for 10 min on overnight air dried sections.
I will be happy to send the other fixative recipe if you want it. 

 

Gayle M. Callis

HTL(ASCP)HT,MT

Bozeman MT 

 

 

 

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From pruegg <@t> ihctech.net  Sat May 30 11:48:41 2009
From: pruegg <@t> ihctech.net (Patsy Ruegg)
Date: Sat May 30 11:48:47 2009
Subject: [Histonet] Microtome Problems
In-Reply-To: 
References: 
	
Message-ID: <6564031D64524FE9B3D415EBC13C62D3@prueggihctechlt>

I would get the service guy back and have him make it right, don't just let
them get away with providing this lousy service, make them take
responsibility for their work.
Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net 
www.ihcrg.org


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
mari.ann.mailhiot@leica-microsystems.com
Sent: Friday, May 29, 2009 10:50 AM
To: Veterinary Services Laboratory
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Microtome Problems

HI Sharon

I am going to refer your email to my collegues in Germany. They will be
able to get you the help you need and all the information..

Best Regards

Mari Ann Mailhiot BA HT ASCP
Application Specialist/Trainer
Leica Microsystems
Biosystems Division
Technical Assistance Center
800 248 0123 x7267
847 236 3063 fax
mari.ann.mailhiot@leica-microsystems.com
www.leica-microsystems.com


                                                                           
             "Veterinary                                                   
             Services                                                      
             Laboratory"                                                To 
              
             .com>                                                      cc 
             Sent by:                                                      
             histonet-bounces@                                     Subject 
             lists.utsouthwest         [Histonet] Microtome Problems       
             ern.edu                                                       
                                                                           
                                                                           
             05/29/2009 09:11                                              
             AM                                                            
                                                                           





Hello All



I'm new to the group and histo. (3 years) so far I'm getting a lot of
useful
tips from all the messages.  Thank you.



I have two microtomes that were working almost perfectly before I had a
technician service them.  Now, the Leica 2035 Jung biocuts' blade holder is
twisting to the side (going up at one end).  I have tried tightening,
slacking, and cleaning the entire assembly to no avail.  Does anyone know
of
where I might be able to source one or even how to fix the one I have.



Secondly, the Leitz 1512 rotary microtome, is now jumping forward and
giving
a loud clanking noise.  It takes huge chunks out of the tissue.



Any and all help is appreciated,



Thank you

Sharon



Ms. Sharon Drayton

Veterinary Services Laboratory

Ministry of Agriculture & Rural Development

The Pine

St. Michael BB11091

Barbados



Tel:  (246) 427-5492 or (246) 427-5073

Fax: (246)426-7517

Email:  vetlab@caribsurf.com

Website:  www.agriculture.gov.bb





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From pruegg <@t> ihctech.net  Sat May 30 12:15:57 2009
From: pruegg <@t> ihctech.net (Patsy Ruegg)
Date: Sat May 30 12:16:02 2009
Subject: [Histonet] coverslippers
Message-ID: <8111422C08D4484C926A16D166A32605@prueggihctechlt>

I am in the process of helping to set up a new histology lab and we want to
get a stand alone glass coverslipper.  I know that this is one piece of
equipment that can be really finicky.  SurgiPath used to market a small
glass coverslipper which has now been bought by Dako.  Are there any of you
out there that use or used this coverslipper when SuriPath sold it?  I am
wondering how this coverslipper performed in the field?

 

Cheers,

Patsy

 

Patsy Ruegg, HT(ASCP)QIHC

IHCtech

12635 Montview Blvd. Ste.215

Aurora, CO 80045

720-859-4060

fax 720-859-4110

www.ihctech.net 

www.ihcrg.org

 

From kimtournear <@t> yahoo.com  Sat May 30 15:53:27 2009
From: kimtournear <@t> yahoo.com (Kim Tournear)
Date: Sat May 30 15:53:31 2009
Subject: [Histonet] recycled formalin QC
Message-ID: <758874.29778.qm@web54201.mail.re2.yahoo.com>

Is anyone out there using the "slurry" system for recycling commercially prepared 10% NBF? And if so, how?do you know your getting 10% again?
Thanks,


~Kim Tournear?~ HT (ASCP), QIHC (ASCP)
Histology Supervisor
Tucson Medical Center
Tucson,? AZ
~Don't?let your life end before it begins~
OU Rocks!!!! (that would be Oklahoma....)


      
From AnthonyH <@t> chw.edu.au  Sun May 31 18:10:48 2009
From: AnthonyH <@t> chw.edu.au (Tony Henwood)
Date: Sun May 31 18:11:01 2009
Subject: [Histonet] muscle striations
In-Reply-To: <7722595275A4DD4FA225B92CDBF174A18C50DBF4E0@EXC-MBX3.cfs.le.ac.uk>
Message-ID: 

Richard,

Cherukian's Phosphotungstic Acid Haematoxylin works well

Principle:	
Cherukian's modification employs an eosin solution that stains the erythrocytes red and differentiates them from the blue fibrin.

Fixation:      10% buffered formalin.
Microtomy:     paraffin sections at 5?m.

Controls:	
Use brain sections and section of muscle.  A good stain will demonstrate the dendrites as blue where as in a bad stain they appear light grey to salmon in colour.  Nuclei, fibrin, platelets and muscle will be blue, red cells and collagen appear red. Muscle striations should be well defined.

Reagents: 

1.	Stock Eosin:   - Warning: Flammable - see MSDS
           Eosin Y, water soluble (CI 45380)       0.5g
           Distilled Water                         	  10ml
           80% Ethanol                             	190ml 

2.	Working Eosin Solution:
Stock Eosin		10ml
Before use add 50ul glacial acetic acid.

3.	1% Periodic Acid

4.	PTAH solution
            Haematoxylin (CI 75290)       	   0.5g
            Phosphotungstic Acid           		    10g
            Distilled water               		500ml

Dissolve solid ingredients in separate portions of the water.  Use gentle heat for Haematoxylin.  Combine solutions when cool.  Add 0.088g potassium permanganate to ripen.  The stain is ready to use.

 

Procedure:

1.	Dewax and hydrate sections to 80% alcohol.
2.	Place slides in working eosin solution for 30 seconds.
3.	Wash slides in distilled water for a few seconds.
4.	Place slides in 1% periodic acid for 20 minutes.
5.	Wash slides in water for 3 minutes.
6.	Place slides in PTAH solution for 30-90 minutes in 60oC oven.  Check from 30 minutes on.
7.	Dehydrate, clear and mount.



Results: 
Dendrites, nuclei, fibrin, platelets and muscle - 	blue
Red blood cells and collagen - 			red.


Notes: 

Reference:	 

	1.	, C.J., Histologic. 8(4); 105, (1977).
	2.	Luna, L., Histologic. 5(2); 66, (1975).

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Edwards, R.E.
Sent: Thursday, 28 May 2009 6:29 PM
To: Histonet
Subject: [Histonet] muscle striations



Any favourite methods for the above??  I'm aware of Heidenhain's and Mallory's PTAH.
                           Many thanks
                              Richard Edwards
                                    University of Leicester

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From AnthonyH <@t> chw.edu.au  Sun May 31 18:51:33 2009
From: AnthonyH <@t> chw.edu.au (Tony Henwood)
Date: Sun May 31 18:51:38 2009
Subject: [Histonet] Peroxidase block again................
In-Reply-To: <8CBAE66781E438A-38C-1C80@FWM-M20.sysops.aol.com>
Message-ID: 

Yep,

You can use alkaline phosphatase labelling instead of peroxidase

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
njoydobro@aol.com
Sent: Friday, 29 May 2009 9:20 PM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Peroxidase block again................


good morning,

???? I would like to pose this questions again...Is there anyone out
there who has eliminated using Peroxidase Block on either IHC or ICC?
Your feedback is very helpful.


Thanks,
Gene
Cleveland Clinic _______________________________________________
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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 note also confirms that this email message has been
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From ooi.ting.huay <@t> nhc.com.sg  Sun May 31 20:23:04 2009
From: ooi.ting.huay <@t> nhc.com.sg (ooi.ting.huay@nhc.com.sg)
Date: Sun May 31 20:24:50 2009
Subject: [Histonet] problem with attaching MMA section to glass slide
Message-ID: 

To all histonetters,

I am currently working with the MMA embedded section. I need to attach 
about 100micro thick sample to glass slide prior to staining. The sample 
is direct from saw microtome. Secure attached is needed so that the 
section will not detach from glass slide while staining. 

I have tried the Haupts adhesive coated slides and bond-rite slides. It 
doesnt work very well...

Please advice and share your expertists. Any suggestion and feedback are 
welcome. Thank you very much!



Regards,
Ooi
_________________________________________________ 
Confidential information. Unauthorized use or disclosure prohibited.
Refer http://www.singhealth.com.sg/ContactUs/#Disclaimer
From jim.manavis <@t> imvs.sa.gov.au  Sun May 31 21:55:50 2009
From: jim.manavis <@t> imvs.sa.gov.au (Jim Manavis)
Date: Sun May 31 21:57:08 2009
Subject: [Histonet] MAP-2 antibody
In-Reply-To: <4A3619571D9F6C4CB79C980E91DBE4E60106801E@TNTRIEXEVS03.triadhospitals.net>
References: <4A3619571D9F6C4CB79C980E91DBE4E60106801E@TNTRIEXEVS03.triadhospitals.net>
Message-ID: <003e01c9e264$78bfea20$2c6c140a@41984n>

Robert

If you are using formalin fixed paraffin embedded human material, Sigma have
a very nice mouse monoclonal that works beautifully. The cat no. is M-4403
(clone HM-2).

Cheers

Jim

Jim Manavis
Laboratory Manager
Hanson Institute
Centre for Neurological Diseases
IMVS, Adelaide, SA, 5000
Australia
Phone: 61-08-8222-3668 / 0401120697
FAX: 61-08-8222 3392
email: jim.manavis@imvs.sa.gov.au
Disclaimer: Not this little black duck! 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lott, Robert
Sent: Saturday, 30 May 2009 2:05 AM
To: histonet@lists.utsouthwestern.edu
Cc: Christa Hladik
Subject: [Histonet] MAP-2 antibody

Hi Everyone,

One of our pathologists is interested in MAP-2 IHC for the following
application:

 

Microtubule Associated Protein-2 Is a Sensitive Marker of Primary and
Metastatic Neuroblastoma.                         

Krishnan C, Heerema-McKenney A, Arber DA.  Department of Pathology,
Stanford University, Stanford, CA.

 

Comment: Microtubule associated protein-2 (MAP-2) is a protein expressed
in high levels in cells derived from the neural crest.  

It is a very robust marker of neuronal differentiation and has been used
most often in evaluating neuronal differentiation in tumors 

of the central nervous system.  MAP-2 showed comparable sensitivity in
staining primary neuroblastomas 

(pre- and post-treatment samples) as compared to synaptophysin,
chromogranin, CD56, and beta-catenin.  

In contrast to other markers of neuroblastoma, MAP-2 did not show
significant cross reactivity to native bone marrow 

precursors, thus eliminating a potential source of confusion.

 

Does anyone use this antibody for this or like applications....

I have looked and am "fairly" sure that there is not an IVD available,
but would still be interested in knowing which antibody you use.

 

Thanks,

 

Robert L. Lott, HTL(ASCP) / Manager, Anatomic Pathology 

Trinity Medical Center/ LabFirst/ 800 Montclair Road / Birmingham, AL
35213/ 205-592-5387 

 



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