[Histonet] thionin staining
Dear all, I have problems with thionin staining. I am using this on paraffin slices of hypothalami. In the past, I never experienced problems, but the last times, the staining is washed off by going through the ethanol after staining (50%, 70%, 90%, 100%). I tried already a few protocols, but every time the same result. Does anybody have experience with it? regards An Eerdekens ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cre protocol-fresh frozen section
Good Afternoon, I would like to do fluoresent staining for Cre expression in thymus. Does anyone have protocol for doing this. I know Covance sells a Cre antibody. Best regards, James ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] CLIA classes?
I would be interested in this as well. I supervise a dermpath lab and have for five years. I have always passed with no deficiencies on my inspections. The inspector always tells me that I document to much stuff. Thanks, Erika -Original Message- From: Sands, Jenn jsa...@innovishealth.com Sent: Thursday, February 3, 2011 11:02am To: indytreeg...@sbcglobal.net, Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] CLIA classes? I would be interested in this information as well. Thank you. Jenn Sands, CT(ASCP)cm, HTL(ASCP)cm -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of indytreeg...@sbcglobal.net Sent: Thursday, February 03, 2011 9:56 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] CLIA classes? Hi all, Does anyone know of a class available for learning about CLIA compliance and regulations, specifically for a Mohs/Dermpath lab? Thanks for any advice you can give me on this! Lyn L. Treeger, B.S.HT(ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any 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
[Histonet] Thionin staining
An - You didn't mention what cells you were staining, but I assume it's mast cells. Thionin stains are very pH dependent. For mast cells, the pH should be 1. At that pH only mast cells stain. Cheryl Cheryl Crowder, BA, HTL(ASCP) Crowder Histology Consulting 4952 Alvin Dark Ave. Baton Rouge, LA 70820 (225) 772-2865 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Histonet Digest, Vol 87, Issue 7
Patty view utube if you can. --Original Message-- From: histonet-requ...@lists.utsouthwestern.edu histonet-requ...@lists.utsouthwestern.edu Date: Fri Feb 04, 2011 -- 04:58:02 AM To: histonet@lists.utsouthwestern.edu Subject:Histonet Digest, Vol 87, Issue 7 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@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. Formaldehyde monitor (Bob Nienhuis) 2. Consult accessioning question (Richard Cartun) 3. Re: Consult accessioning question (Victor Tobias) 4. RE: Consult accessioning question (wanda.sm...@hcahealthcare.com) 5. Re: Formaldehyde monitor (kim.dona...@bhcpns.org) 6. RE: Formaldehyde monitor (wanda.sm...@hcahealthcare.com) 7. RE: Consult accessioning question (Mike Pence) 8. Re: Off Topic: researchers very funny (Merced M Leiker) 9. RE: Consult accessioning question (Weems, Joyce) 10. TTF-1/background staining (Paula Lucas) 11. Microwave processing (Webb, Dorothy L) 12. FW: [Histonet] Formaldehyde monitor (Vanessa Avalos) 13. Re: Microwave processing (Rene J Buesa) 14. Temporary histology position in San Diego Ca. (James Watson) 15. RE: TTF-1/background staining (Debra Siena) 16. RE: Her2 Fixation Requirement (Rathborne, Toni) 17. Karnovsky and Roots stain (Nicole Cosenza) 18. RE: TTF-1/background staining (Anthony Reilly) 19. Re: TTF-1/background staining (Mark Tarango) 20. thionin staining (An Eerdekens) 21. Cre protocol-fresh frozen section (James Dooley) 22. RE: CLIA classes? (er...@mapslab.net) -- Message: 1 Date: Thu, 3 Feb 2011 10:29:18 -0800 From: Bob Nienhuis bob.nienh...@gmail.com Subject: [Histonet] Formaldehyde monitor To: histonet@lists.utsouthwestern.edu Message-ID: AANLkTimcGcnxpOpCQHop5WwxXPyvu1Mwo=ukanjsq...@mail.gmail.com Content-Type: text/plain; charset=ISO-8859-1 Anyone know of a cheap formaldehyde gas monitor or concentration test method? I found a couple of monitors on Google, but they seem to run abour $1500+ Bob Sepulveda VA Med Center Los Angeles -- Message: 2 Date: Thu, 03 Feb 2011 13:43:05 -0500 From: Richard Cartun rcar...@harthosp.org Subject: [Histonet] Consult accessioning question To: Histonet histonet@lists.utsouthwestern.edu Message-ID: 4d4ab0e8.7400.007...@harthosp.org Content-Type: text/plain; charset=US-ASCII When accessioning a case for consultation, do you use the date on the paperwork from the referring hospital/pathologist as the new Date-of-service? If so, what do you do when the paperwork gives a date (say Friday) and then the specimen is not sent out until the following Monday? Thank you. Richard Richard W. Cartun, MS, PhD Director, Histology Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax -- Message: 3 Date: Thu, 03 Feb 2011 10:59:53 -0800 From: Victor Tobias vic...@pathology.washington.edu Subject: Re: [Histonet] Consult accessioning question To: histonet@lists.utsouthwestern.edu Message-ID: 4d4afb29.8070...@pathology.washington.edu Content-Type: text/plain; charset=ISO-8859-1; format=flowed Richard, We use the date of accessioning as the DOS and we wouldn't accession it until we have received the specimen/materials. Victor Victor Tobias HT(ASCP) Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 vic...@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. On 2/3/2011 10:43 AM, Richard Cartun wrote: When accessioning a case for consultation, do you use the date on the paperwork from the referring hospital/pathologist as the new Date-of-service? If so, what do you do when the paperwork gives a date (say Friday) and then the specimen is not sent out until the following Monday? Thank you.
RE: [Histonet] TTF-1/background staining
Hugh thanks for your reply and I'll type my answers in blue below after the questions: _ From: Hugh Luk [mailto:hlu...@msn.com] Sent: Thursday, February 03, 2011 6:21 PM To: plu...@biopath.org Subject: RE: [Histonet] TTF-1/background staining Paula, Incomplete IHC staining has many factors. 1) What type of antigen retrieval are you using? We use the Lab Vision's Citrate Buffer Ph 6 Can you use a high pH solution instead? Unmasking hard epitopes (TTF-1 can be stubborn) will look better under a higher pH AR (or TR for Dako users). Are you re-using your solution? Please say No NO or at least not often. Lastly how are your retrieving? We use Biocare's Decloaking Chamber, basically a pressure cooker Pressure HIER or rolling boil methods work best for TTF1. 2) What type of buffer are you using. We use TBS with Tween added: a small capful to a 5 liter container TBS or PBS should both work. Is it pH'd correctly? Since you automate, does your buffer contain Tween 20? Too much or too little could ruin your day. 3) Primary antibody. Concentrate? It's a predilute Could your diluent (solution) be faulty or your titre incorrect? Call or email Cell Marque to help you with this? I did call Cell Marque and they gave me some suggestions such as using an EDTA based retrieval solution and to reduce the antibody time to 15 minutes, and to also increase the blocker from 5 to 10 minutes Another email from Tony asks about dilution too, and hints at problems with the anitbody (AR). I use Dako's version of TTF-1 for human tissues, and I have not seen a problem. Perhaps you could call Dako and procure a free sample? I'll try that 4) Fixation. TTF-1 isn't known to be fixation-dependent, but how long are your tissues sitting in formalin before processing? It depends.we receive samples from the hospital, and so it can vary. The smaller samples are in formalin at least 6 hours: includes the sample sitting in the formalin container waiting for gross, plus then the processor time Problems with over or under fixation could cause your description. Also, what fixative are you using? We use 10% Formalin I have heard various fixatives being used on liver biopsies: Carnoy's, Helly's, Hollende's, Zinc formalin, ect. If you are using another fixative, can you go back to 10% neutral buffered formalin? 5) Tissue processor. How is your tissue processor? It's a VIP and the paraffin is checked at 60 degrees Could your paraffin be overheating? 6) Detection system. I have simply never tried your kit, but I will say several antibodies from Thermo have worked well. Thermo lists their polymer system as non-avidin-biotin based, and should be ultra sensitive. If it works on everything else (especially stains that visualize the nucleus), I would say your kit is not the problem. But it still might not like your antibody. For example, the nuclear epitope is rather small and Dako's polymer kit (Envision plus) was a large bio-synthesized molecule that many people thought would sterically hinder a good percentage of binding, reducing the signal. Don't know if that's true, it works okay for us. 7) 30 minute primary incubation should be fine. Is there any chance your slides are drying while incubating? I'm not 100% sure..the slides look to me like they are not drying out. We apply the buffer as soon as we place the slides on the machine, and then we keep the cover to the machine closed while it's running, so I'm assuming the slides aren't drying out We sometimes put paper towels on the bottom of the stainer to assure the chamber is not drying out our slides (but that's only when we get REALLY paranoid). 8) Liver. Is dirty. Debbie has a point that the liver may be working against you. You could try a protein block, or something that Thermo may suggest to deter background. Do you ink your liver biopsies? No we don't ink the liver bx's Davidson's inks used to give us some problems with IHC on small tissues (so we went to marking with eosin or hematoxylin). 9) Liver does not normally stain TTF-1. HCC's should be negative. Lung would stain well (unless your Lung tissue is also a metastisis), but for example breast mets could be focal and weak to diffuse and strong. It depends on the tumor. Could the tumor be of this type? I will ask the pathologist this question Good luck and feel free to email me if something I wrote needs to be clarified. I apologize for the rambling on... don't apologize! I really appreciate this Hugh Cancer Research center of Hawaii From: dsi...@statlab.com To: plu...@biopath.org; histonet@lists.utsouthwestern.edu Date: Thu, 3 Feb 2011 16:02:22 -0600 Subject: RE: [Histonet] TTF-1/background staining CC: Hi Paula, I am probably reading between the lines here but on your TTF protocol do you use a streptavidin biotin detection system and if so do you block for endogenous biotin, liver will have endogenous biotin where lung may not have as much? thanks Debbie
[Histonet] Bond Refine Red for manual IHC
Hello all, I wonder if anyone is using the Bond Polymer Refine Red Detection kit (cat # DS390, Leica) for manual IHC. I've seen some beautiful staining, the technical specialist at Leica, however, told me it is only for Leica automatic stainers. It is an AP stain. Has anybody tried it? Thanks a lot, Sarka Lhotak McMaster University Hamilton, Ontario, Canada ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Bond Refine Red for manual IHC
Yes, we've used both the AP Red and DAB Refine kits manually with comparable results to the automation Ronnie Houston Anatomic Pathology Manager Nationwide Children's Hospital Columbus OH 43205 (614) 722 5450 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sarka Lhotak Sent: Friday, February 04, 2011 10:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bond Refine Red for manual IHC Hello all, I wonder if anyone is using the Bond Polymer Refine Red Detection kit (cat # DS390, Leica) for manual IHC. I've seen some beautiful staining, the technical specialist at Leica, however, told me it is only for Leica automatic stainers. It is an AP stain. Has anybody tried it? Thanks a lot, Sarka Lhotak McMaster University Hamilton, Ontario, Canada ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Confidentiality Notice: The following mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Looking for someone with muscle biopsy experience in NY
Dear Histonet Members, I hope you are doing well. I am a Recruiter at a highly successful and well respected Healthcare recruiting firm. I help place Lab Professionals in permanent positions across the country and I wanted to see if you are interested in exploring other career opportunities? We are completely free of charge to candidates and and we work on quite a few laboratory openings across the country. Our clients typically assist with relocation expenses. I am currently working on 2 open positions with a fast paced and service-oriented company in New York, NY. This company is an innovative, commercial laboratory that specializes in performing and developing testing services that serve the technically advanced medical community with a focus on neurological disorders. They are looking to hire on for the following positions: * Experienced Histotech - NYS licensed histotech, 5+ years experience, IHC experience a plus * Histology Supervisor - NYS licensed Clinical Technology Supervisor, 5+ years experience, IHC a must * Muscle/Nerve Technician - Must have muscle biopsy experience They are offering an exceptional compensation package, including health, dental, life, and a 401K plan. They are expanding and looking to hire as soon as possible! If you're interested in learning more about these opportunities or opportunities in a certain geographic location please reply with an updated resume and let me know when a good time to reach you is. If this is not the right fit for you please let me know who you can recommend and give me an idea of what types of positions you'd be interested in hearing about in the future. I cover the entire US and have am working on Lab positions at all levels. We offer a very generous referral bonus for anyone you refer to us that we place into any position across the country. To view some additional opportunities please visit our website at www.ka-recruiting.com. Sincerely, Alisha (Taylor) Dynan, Founder K.A. Recruiting, Inc. Your Partner in Healthcare Recruiting 10 Post Office Square 8th Floor SOUTH Boston, MA 02109 P: (617) 692-2949 F: (617) 507-8009 ali...@ka-recruiting.com www.ka-recruiting.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histology Training Specialist Based in the Chicago Area
Good Morning Histonet! We currently have a very strong Histology Training Specialist opportunity with a World Leader in Histology equipment. This position would be based out of the company’s Corporate Office in the Chicago area and offers an outstanding opportunity for career growth. Please e-mail resumes to m...@personifysearch.com Thanks! Matt Ward *Account Executive* *Personify* 5020 Weston Parkway Suite 315 Cary NC 27513 (Tel) 800.875.6188 direct ext 103 (Fax) 919.460.0642 www.personifysearch.com *Histology Training Specialist * *The Company:* Our client is a leading developer and producer of innovative high-tech precision optics systems for the analysis of microstructures. As one of the market leaders in each of the fields of Microscopy, Confocal Laser Scanning Microscopy, Imaging Systems, Specimen Preparation and Medical Equipment. Comprising nine manufacturing facilities in seven countries, sales and service companies in 20 countries and an international network of dealers, the company is represented in over 100 countries. *The Opportunity:* The company currently has an opening for a Histology Training Specialist. All applicants must not be adverse to travel, as this is a position that may require you to travel when necessary. Base: Based on Experience Other: Full benefits - 401k program/matching *Primary Responsibilities:* The primary responsibility of this role will be to provide technical phone support by answering questions, troubleshooting problems, logging and closing complaint files and escalating major issues to appropriate company personnel. This role will also provide technical training on specified products in the company's newly constructed state-of-the-art Customer Support Laboratory. Training programs are designed for small groups to ensure maximum customer learning and satisfaction. Additional Responsibilities: - Provide product and applications phone support to end-users, field personnel and dealers for all product lines - Log all calls into Customer Support Database - Participate in development of training materials and conduct classes and labs for customers, employees and others as needed - Serve as technical liaison to Customer Service/Field Service/Product Management departments *Education and Experience Required:* Ability to interact with various people in a calm and positive fashion and the ability to effectively communicate information to groups of participants is required. Experience with data entry, MS Office programs (PowerPoint, Lotus Notes, Word) is also required. HT/HTL/QIHC (ASCP) is helpful but not required. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Weekly reminder
Hi Georgia, Alabama, ALL histotechs, The Georgia Society for Histotechnology invites you to our meeting March 25-27, 2011 at Callaway Gardens in Pine Mountain, Georgia which is near Columbus, Ga. and very convenient to Alabama folks, so come across the line. The invitation extends to any other states as well. Callaway Gardens is a fantastic site for family vacations, golf lovers, nature lovers, so come to Georgia for a visit and take in a wealth of histology knowledge. The deadline for making hotel reservations is March 1, 2011 so that gives you a month to make your plans to attend, don't delay. The Mountain Creek Inn, Callaway Gardens, Pine Mountain, Georgia is the location and you can call for hotel reservations at 1-800-225-5292. Room rates start at $99 which includes Continental Breakfast and Admission to the Park. For more information about things to do at Callaway click on the link here: http://www.callawaygardens.com/resort/things-to-do/georgia-fun.aspx Our theme this year is METAMORPHOSIS: Transforming Histotechs. The complete program can be downloaded from our website at this link: www.histosearch.com/gshhttp://www.histosearch.com/gshhttp://www.histosearch.com/gsh%3chttp:/www.histosearch.com/gsh then click on GSH symposium link at the bottom of the home page. There you will find the complete program with registration form. The vendor registration form is on the same page for any last minute vendors who want to exhibit at our meeting. If anyone has questions, please contact me for assistance. Come TRANSFORM yourselves. Shirley Powell GSH Secretary Shirley A. Powell, HT(ASCP)HTL, QIHC Technical Director Histology Curricular Support Laboratory Mercer University School of Medicine 1550 College Street Macon, GA 31207 478-301-2374 Lab 478-301-5489 Fax ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] basic start-up fluorescence microscope
Hello, Can anyone please recommend a basic laboratory start-up fluorescence microscope ( DIC a must; green and red channels would be nice) ? Kind regards, Marti ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] CLIA classes?
Anderson Education offers a good course on this for CEU's. You can contact them at 1-800-532-2332 Kim Donadio Pathology Supervisor Baptist Hospital 1000 W Moreno St. Pensacola FL 32501 Phone (850) 469-7718 Fax (850) 434-4996 er...@mapslab.net Sent by: histonet-boun...@lists.utsouthwestern.edu 02/04/2011 04:55 AM To Sands, Jenn jsa...@innovishealth.com cc histonet@lists.utsouthwestern.edu Subject RE: [Histonet] CLIA classes? I would be interested in this as well. I supervise a dermpath lab and have for five years. I have always passed with no deficiencies on my inspections. The inspector always tells me that I document to much stuff. Thanks, Erika -Original Message- From: Sands, Jenn jsa...@innovishealth.com Sent: Thursday, February 3, 2011 11:02am To: indytreeg...@sbcglobal.net, Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] CLIA classes? I would be interested in this information as well. Thank you. Jenn Sands, CT(ASCP)cm, HTL(ASCP)cm -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of indytreeg...@sbcglobal.net Sent: Thursday, February 03, 2011 9:56 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] CLIA classes? Hi all, Does anyone know of a class available for learning about CLIA compliance and regulations, specifically for a Mohs/Dermpath lab? Thanks for any advice you can give me on this! Lyn L. Treeger, B.S.HT(ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any 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 - All electronic data transmissions originating from or sent to Baptist Health Care Corporation (BHC) are subject to monitoring. This message along with any attached data, are the confidential and proprietary communications of BHC and are intended to be received only by the individual or individuals to whom the message has been addressed. If the reader of this message is not the intended recipient, please take notice that any use, copying, printing, forwarding or distribution of this message, in any form, is strictly prohibited and may violate State or Federal Law. If you have received this transmission in error, please delete or destroy all copies of this message. For questions, contact the BHC Privacy Officer at (850) 434-4472. Rev.10/07. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Her2 Fixation Requirement
We adhere the ink with bouins sol'n. Have had no problems. Kim Donadio Pathology Supervisor Baptist Hospital 1000 W Moreno St. Pensacola FL 32501 Phone (850) 469-7718 Fax (850) 434-4996 Rathborne, Toni trathbo...@somerset-healthcare.com Sent by: histonet-boun...@lists.utsouthwestern.edu 02/03/2011 04:54 PM To Weems, Joyce jwe...@sjha.org, Paula Lucas plu...@biopath.org, histonet@lists.utsouthwestern.edu cc Subject RE: [Histonet] Her2 Fixation Requirement Does anyone have issues with inking the specimen after it has been placed in formalin? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu]On Behalf Of Weems, Joyce Sent: Wednesday, February 02, 2011 12:59 PM To: Paula Lucas; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Her2 Fixation Requirement We have the nurse document the time of removal on the requisition. The next requisition update will have a spot for this. They have been very cooperative and do a good job. In addition, you must document the cold ischmic time - that is the time from removal until time in formalin. This is important when the specimen goes for xray or whatever. So there is a removal time, an into formalin time and an out of formalin time. Then if we don't have time allowed to meet the fixation time with the regular, it is put on a late processor, if we have one available, or it is held overnight. And if it has to come off on Sunday, we have a med tech remove it from the processor and it waits for us to come on Monday to embed it. The pathologists document all this time in the report. Best! 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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Paula Lucas Sent: Wednesday, February 02, 2011 12:11 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Her2 Fixation Requirement Hello histoland I was just given the task to find a solution that is easy and will also comply with the CAP guideline for formalin fixation documentation, and so I started my research on the Histonet archives. I found some good information, but was hoping to get more feedback. Would you mind sharing with me the actions you are taking to comply with the guideline? We are a private lab and we provide histology/pathology service for 2 hospitals and a few surgery centers. We send our blocks to Genzyme for Her2, and we must document on their test order sheet how many hours the tissues have been fixed in formalin. I'm assuming I will need to start keeping a log here, with documentation that shows what time the tissue was excised and placed in formalin from the OR, and then documentation that shows the time it was dissected and then placed in the tissue processor. The problem that I may come across is getting the OR nurse to document the time for us. I don't know...maybe we need to put another sections on our requisition form, or maybe something on the formalin container itself for the nurse to write on. It'll be a hassle at first but if I can get the hospitals lab director involved, I'm sure it will work itself out. Anyway, if you wouldn't mind sharing some of your ideas, I would really appreciate it. Paula Lucas Lab Manager Bio-Path Medical Group Fountain Valley, CA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This e-mail, 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 delete this message, and reply to the sender regarding the error in a separate email. ___ 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.
[Histonet] CE
I'm looking for some cost effective CE for my staff. Does anyone know if the CAP still has TechSamples? Please email at andrea.con...@atlanticare.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] thionin staining
If this is for Nissl staining, the pH of the thionine solution should be 3.5 to 4 and the following rinses should be no more acidic than that. Distilled water is suitable. Alcohol-water mixtures generally remove dyes to a greater extent than 100% alcohol. I shake the washed slides to get rid of most of the water, then go straight to the first of 3 changes of 100% ethanol. Almost no blue is lost from the sections. The first alcohol isn't 100% any more after one or two batches of slides have gone through, so for large numbers of slides it is more economical to go through 95% (quickly) then 3X100%. If you use 50% and 70% alcohol you can expect dye to be extracted from the stained sections. It's not unusual to get unsatisfactory batches of thionine, and for at least one purpose, showing the canaliculi and lacunae of bone, the late Russ Allison found that proper staining could be obtained only with batches of thionine that had been certified by the Biological Stain Commission. See Allison RT (1995) Picro-thionin (Schmorl) staining of bone and other hard tissues. Brit. J. Biomed. Sci. 52: 162-164. The B.S.C.'s tests for thionine are a mast cell stain and a stain for plant tissue infected with a fungus. The dye must also meet spectrophotometric criteria. See Penney DP (2002) Analysis and testing of biological stains - the Biological Stain commission Procedures. Biotech. Histochem. 77:237-275. A minor revision to the criteria for thionine was published in 2008: Lyon HO Kiernan JA (2008) Notes from the Biological Stain Commission. Biotech. Histochem. 83(5):285-288. Make sure your dye really is thionine (CI 52000). There is a dye called thionine blue (CI 52025, Basic blue 25) that is not a substitute. See Conn's Biological Stains or Bryan Llewellyn's StainsFile http://stainsfile.info/StainsFile/dyes/52000.htm . Finally, the word thionine is commonly mis-spelled, without its terminal e. The e should be there because thionine is an amine, in contrast to eosin, which is not. Dictionaries (English or US) give the correct spelling. John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: An Eerdekens an.eerdek...@med.kuleuven.be Date: Friday, February 4, 2011 3:21 Subject: [Histonet] thionin staining To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Dear all, I have problems with thionin staining. I am using this on paraffin slices of hypothalami. In the past, I never experienced problems, but the last times, the staining is washed off by going through the ethanol after staining (50%, 70%, 90%, 100%). I tried already a few protocols, but every time the same result. Does anybody have experience with it? regards An Eerdekens ___ Histonet mailing list Histonet@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] veterinary IHC for Ornithobacterium
Does anyone do IHC staining for Ornithobacterium rhinotracheale (ORT), a respiratory pathogen in poultry? I'm trying to find a lab to do it, or a vendor source for antibody. Much thanks in advance, Jan Shivers Senior Scientist Histology/IHC/EM Section Head Pathology Teaching Program University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive...@umn.edu (Confidentiality Notice: This message, together with any attachments, is intended only for the use of the individual or entity to which it is addressed and may contain confidential or privileged information. If you think you have received this message in error, please advise the sender and then delete this message and any attachments immediately.) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HCN4 staining
Hello, I'm hoping somebody out there can give me some help with HCN4 staining, I was asked to stain HCN4 on mouse heart and it needs to stain the SA node. It is very hard to find that area since the node is very small. I have tried serial sections but not sure I hit the area or if I'm having problems with the stain. The antibody is Rat monoclonal Secondary Alexa Flour 488 goat anti-rat Counter stain is Hoechst I tried frozen and paraffin sections with no luck Any suggestions will help greatly Thanks, Sharon ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Acetylcholinesterase
Nicole Cosenza asks about acetylcholinesterase methods. Years and years ago in London we used Koelle's method to demonstrate cholinesterase in muscle and mouse diaphragms. The method we followed was in Pearse E, Histochemistry - Theoretical and Applied. Volume 2. Churchill Livingstone. London 1972. Pages 1312 - 1316 has several different methods. The enzyme was visualized with ammonium sulphide. The tissues were mounted in glycerine jelly. If the method worked too well and we could not see the motor end plates, we adjusted the pH to reduce staining. Dr Filipe went on to publish her own method in Filipe I., Lake B., Histochemistry in Pathology. Churchill Livingstone. London 1983 page 322. In her method, she used osmium to visualise the enzyme, instead of ammonium sulphide. Stain technology used to have articles about the method too. I have not heard of Karnovsky or Roots methods, but lot of different methods were published in the early days of enzyme histochemistry. Hope this helps Michael Titford Pathology - USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Artifact or dirt on slides using a tape cover slipper?
At our facility with have 2 Tissue-Tek automated tape cover slippers. The brand of tape that we use is made by a company called Klinipath, KP Tape (dist. By Mercedes Medical). On occasion we get complaints that the slides appear to have areas of dirt or dust on them. It appears to be on the inside. Has anyone else that uses a tape cover slipper ran across this particular problem? If so, what did you do to troubleshoot? If it is the tape, is there another brand or type that is preferable? Nancy Mitchell Pathology Arts, Inc Director of Sales and Marketing 951-270-0605 909-732-1666-Cell na...@pathologyarts.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Artifact or dirt on slides using a tape cover slipper?
Nancy, We get areas that appear brown when the tape doesn't lay flat or adhere to the slide. Most of the time this happens on decal slides or hard tissue where the tissue may be lifted a little. Try recoverslipping. Also, if it's not happening on these types of tissue, you may need more xylene dispensed. Sometimes, we just have to resort to the old-fashioned way of coverslipping with mounting media. I've never used the Klinipath tape. I prefer to stick with Sakura's brand - don't want to take any chances! Laurie Colbert -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Nancy Sent: Friday, February 04, 2011 12:12 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Artifact or dirt on slides using a tape cover slipper? At our facility with have 2 Tissue-Tek automated tape cover slippers. The brand of tape that we use is made by a company called Klinipath, KP Tape (dist. By Mercedes Medical). On occasion we get complaints that the slides appear to have areas of dirt or dust on them. It appears to be on the inside. Has anyone else that uses a tape cover slipper ran across this particular problem? If so, what did you do to troubleshoot? If it is the tape, is there another brand or type that is preferable? Nancy Mitchell Pathology Arts, Inc Director of Sales and Marketing 951-270-0605 909-732-1666-Cell na...@pathologyarts.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
Re: [Histonet] Karnovsky and Roots stain
Karnovsky Roots is (IMHO) the best histochemical method for choline esterase activity. In muscles, acetylcholinesterase (AChE) is the only such esterase shown by this method, and it is in the subneural apparatus of the motor endplate. Some counterstains (notably silver methods for the innervating axons) can remove the brown copper ferrocyanide product. Another way to show motor endplates is with a method that picks up all esterases. In muscle, the endplate AChE shows up sooner than the enzymes present in all cells. Indigogenic esterase methods can be followed by silver staining of axons. Why do you need or want to use fresh frozen, unfixed tissue sections? This makes no sense in the world of esterase activity histochemistry. There haven't been any developments in this field since the 1960s other than labelled alpha-bungarotoxin and immunohistochemistry. An inexpensive book is Van Noorden CJF Frederiks WM 1992. Enzyme Histochemistry. Oxford Univ Press and Royal Microscopical Soc. ISBN0198564341. Another one is Lojda, Gossrau Schiebler 1979. Enzyme histochemistry. Berlin: Springer. ISBN 0387092692. A quick web search indicates that both are available and cost less than $10 second-hand. John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: Nicole Cosenza ncose...@siumed.edu Date: Thursday, February 3, 2011 18:45 Subject: [Histonet] Karnovsky and Roots stain To: histonet@lists.utsouthwestern.edu I am looking into a project involving motor end plate staining. Literature that I've found continually references Karnovsky and Roots from the 60s. However the papers are not supplying all the details. Does anyone do AchE staining by this method on fresh frozen, unfixed tissue sections? If so, can I get a more detailed protocol (fixation steps, washes, etc)? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Karnovsky and Roots stain
We do ACH staining on rectal biopsies to evaluate for ganglion cells for Hirschsprung's disease. Our stain is performed on fresh frozen tissue and I believe it's the Karnovsky method, but I'm not sure. Feel free to email if you are interested in out procedure. Lisa M. Van Valkenberg, B.S., HT- ASCP Histology Manager 2300 Children's Plaza Chicago, IL 60614 773-868-8949 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Friday, February 04, 2011 3:15 PM To: Nicole Cosenza Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Karnovsky and Roots stain Karnovsky Roots is (IMHO) the best histochemical method for choline esterase activity. In muscles, acetylcholinesterase (AChE) is the only such esterase shown by this method, and it is in the subneural apparatus of the motor endplate. Some counterstains (notably silver methods for the innervating axons) can remove the brown copper ferrocyanide product. Another way to show motor endplates is with a method that picks up all esterases. In muscle, the endplate AChE shows up sooner than the enzymes present in all cells. Indigogenic esterase methods can be followed by silver staining of axons. Why do you need or want to use fresh frozen, unfixed tissue sections? This makes no sense in the world of esterase activity histochemistry. There haven't been any developments in this field since the 1960s other than labelled alpha-bungarotoxin and immunohistochemistry. An inexpensive book is Van Noorden CJF Frederiks WM 1992. Enzyme Histochemistry. Oxford Univ Press and Royal Microscopical Soc. ISBN0198564341. Another one is Lojda, Gossrau Schiebler 1979. Enzyme histochemistry. Berlin: Springer. ISBN 0387092692. A quick web search indicates that both are available and cost less than $10 second-hand. John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: Nicole Cosenza ncose...@siumed.edu Date: Thursday, February 3, 2011 18:45 Subject: [Histonet] Karnovsky and Roots stain To: histonet@lists.utsouthwestern.edu I am looking into a project involving motor end plate staining. Literature that I've found continually references Karnovsky and Roots from the 60s. However the papers are not supplying all the details. Does anyone do AchE staining by this method on fresh frozen, unfixed tissue sections? If so, can I get a more detailed protocol (fixation steps, washes, etc)? ___ Histonet mailing list Histonet@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] combined cholinesterase-silver stain
I am looking into staining motor end plates. I've come across this combined cholinesterase-silver stain (reference Pestronk and Drachman, 1978). Based on the date of the paper, I'm wondering what the current technique is for this double staining. Anyone currently doing AchE and axon staining on fresh frozen muscle sections? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] pH Meter
Happy Friday everyone! I am in the market for a new pH Meter, and was wondering if any of you had any preferences. I want to keep it simple as possible for the staff. Also, I don't want to spend a ton of money. One of our fellow histonet subscribers recommended purchasing a pH Meter that did a 2-step calibration verses a 3-step. I've always used a 3-step calibration. Any thoughts. I realize that it must have a good glass electrode, calibrated daily, prior to use, and all the maintenence must be followed to keep it in good working order. If you have any suggestions, please provide the make and model #, and if you have a vendor who provides it such as Fisher or Thermo, that would be an added bonus. Thank you, and have a great weekend! Akemi Allison BS, HT(ASCP)HTL Director Phoenix Lab Consulting E-Mail: akemiat3...@yahoo.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Amylase Digestion for glycogen
Hello, We are looking to switch from malt diastase digestion for glycogen to Amylase digestion. I have the new protocol worked up, but one of the Pathologists I work with would like to have an idea of how many labs out there are using Amylase instead of malt diastase for their PAS/D method. If you use amylase for the PAS/D method, could you please let me know who you are and the institiution? Much appreciated. amysue ruppert Histology lab Marshfield Labs Marshfield WI __ The contents of this message may contain private, protected and/or privileged information. If you received this message in error, you should destroy the e-mail message and any attachments or copies, and you are prohibited from retaining, distributing, disclosing or using any information contained within. Please contact the sender and advise of the erroneous delivery by return e-mail or telephone. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Amylase Digestion for glycogen
div style=font-family: 'times new roman'; font-size: 1= 6pxdivIt is good that the pathologist you work with takes an= interest in histochemical methodology, but when and where did he lear= nelementarybiology?/divdiv br//divdivW= hen I was a schoolboy in the 1950s the biology teacher told us that dias= tase and amylase were different names for the enzyme in saliva and pancr= eatic juice that digests starch. This was also true in my two prec= linical years at medical school (early 1960s). The biochemistry teache= rs told us more about starch and its animal equivalent glycogen, both digestible by amylase. Our early-1960s textbooks told us about Claude = Bernard, the great French physiologist who discovered the properties= of glycogen in the mid-1800s and experimentally demonstrated the func= tion of the liver in glucose metabolism and homeostasis./divd= iv br//divdivAccording to the Sigma catalogue, di astase is now quot;an obsolete synonymfor alpha-amylasequot;.= The same catalogue lists alpha-amylases from many sources, includin= g malt. (They also sell beta-amylases, which would also catalyze = the hydrolysis of glycogen.) Sigma's least expensive alpha-amyl= ase is from human saliva./divdiv br//divdiv= Any amylase will do the job. Go with the cheapest. Human drooli= ngs are free and do not contain enzymes that will digest and solubilize polysaaccharides other than glygogen and starch. This is traditional h= istochemistry, well documented in textbooks and peer-reviewed papers f= or about 100 years./divdiv br//divdivJoh= n Kiernan/divdivAnatomy, UWO/divdivLondon, C= anada/divdiv= = =br /On 04/02/11, b class== namequot;Ruppert, Amysuequot; /blt;ruppe=rt.amy...@marshfieldclinic.orggt; wrote:/divblockq= uote cite=mid:201102050003.p15030nh007461@spamfilt class= iwcQuote style=border-left:#00f 1px solid; padding-lef= t: 13px; margin-left: 0px type=citediv class== mimepart text plainHello,br / We are looking to switch f= rom malt diastase digestion for glycogen to Amylase digestion. I have = the new protocol worked up, but one of the Pathologists I work with wo uld like to have an idea of how many labs out there are using Amylase in= stead of malt diastase for their PAS/D method.br / If you use = amylase for the PAS/D method, could you please let me know whoyouare=andtheinstitiution?br / Much appreciated.br /br = /amysue ruppertbr /Histology labbr /Marshfield Labsbr = /Marshfield WIbr /br / ___ 5F ___ 5F= __br /The contents of this message may= contain private, protected and/or privileged information. If you= received this message in error, you should destroy the e-mail message= and any attachments or copies, and you are prohibited from retaining,= distributing, disclosing or using any information contained within.= Please contact the sender and advise of the erroneous delivery by re= turn e-mail or telephone. Thank you for your cooperation.br / /div/blockquote/div ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Where can Fast Blue (Diamidino compound 253/50) be obtained?
Hi, I've searched every catalog and website for Fast Blue (Diamidino compound 253/50), widely used for neuronal retrograde tracing. One recent paper using this compound is: Porreroa et al Mapping of fluorescent protein-expressing neurons and axon pathways in adult and developing Thy1-eYFP-H transgenic mice, Brain Research, 1345, 59-72 where the authors as many before them cite Dr. Illing GmbH Co. KG, Groβ-Umstadt, Germany as the origin. I have been unable to locate this particular vendor. I've written the authors and am waiting for an answer. In the meanwhile, can anyone on this list help? Kindly, *Per M Knutsen *Department of Physics University of California, San Diego 9500 Gilman Drive, La Jolla, CA 92093-0374 T: +1 858 405 2868 E: pknut...@ucsd.edu W: http://pmknutsen.blogspot.com * * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
