[Histonet] Sakura DRS 601 basket hook and slide basket
Hello Let me first thank the list for the nice help I have always found here. (for exemple with Perkins Biomedical Services and autostainer bottles) I am looking now for spares for Sakura DRS™ 601. 4368 : Basket Hook (3 of them if possible) 4768 : 20-Slide Basket (at least 3 to 6 of them) Best regards ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Thermo Scientific Embedding Centre Histostar
We had one screen replaced early on but no problems since. However, we've had to replace the plastic lids several times and the parts are always on back order. The one that covers the molds breaks right off. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bell, Lynne Sent: Tuesday, January 21, 2014 2:54 PM To: 'Gregoire, Rhonda (MAFRI)'; 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: Thermo Scientific Embedding Centre Histostar I have the Histostar embedding center (purchased 9/2012) and have not had any problems with the touch screen. Lynne Bell, HT (ASCP) Histology Team Leader Central Vermont Medical Center 130 Fisher Road Berlin, VT 05641 802-371-4923 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Gregoire, Rhonda (MAFRI) Sent: Tuesday, January 21, 2014 1:09 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Thermo Scientific Embedding Centre Histostar We are looking at purchasing a Histostar and would like to know if you have had any trouble with the touch screen? I had to replace the touch screen on our Histocenter 3 because we could no longer navigate the menu with all the lines going thru it. Wanted to know if it was just our bad luck or if anyone else has had the same experience with the touch screen. Thanks Rhonda Gregoire, MLT Supervisor, Clinical Pathology Veterinary Diagnostic Services Manitoba Agriculture, Food and Rural Development 545 University Crescent Winnipeg, MB R3T 5S6 phone 204-945-7641 fax 204-945-7646 email rhonda.grego...@gov.mb.camailto:rhonda.grego...@gov.mb.ca ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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. Geisinger Health System utilizes an encryption process to safeguard Protected Health Information and other confidential data contained in external e-mail messages. If email is encrypted, the recipient will receive an e-mail instructing them to sign on to the Geisinger Health System Secure E-mail Message Center to retrieve the encrypted e-mail. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cell Marque Mammoglobin (31A5) New Protocol?
Fellow histotechs, We've been running mammoglobin testing on our Ventana Benchmark XT. However, Cell Marque (our supplier of mammoglobin antibody) has changed the concentration. Our old concentration was 0.12 mg/mL, the new concentration is weaker at 0.04 mg/mL. The antibody is a rabbit monoclonal. We tried running the old protocol with the new concentration, but the results were not adequate (see below). We tried running the protocol given to us by Cell Marque, but theirs is specific towards the Benchmark UltraView and also returned inadequate results. We process our tissue from formalin to paraffin. Our lab uses the iVew DAB detection kit. Our old protocol: Deparaffinization Conditioner #1 (short 8 minute conditioning) Mild CC1 (mild 30 minutes conditioning) Standard CC1 (standard 60 minute conditioning) No enzyme 37C antibody incubation temperature No titration Antibody: Mammoglobin 31A5 at 0.12 mg/mL concentration for 32 minutes A/B Block Hematoxylin counterstain 4 minutes Blueing post counterstain 4 minutes Does anyone else use this product from Ventana/Cell Marque? Does anyone have a working protocol for a weaker antibody titration? Any help would be greatly appreciated! *Abby L. H. Maples, BS, MPH, HTL (ASCP)CM* Histotechnologist, Surgical Pathology Mercy Medical Center 701 10th Street SE Cedar Rapids, IA 52403 Lab Phone: (319) 398-6827 Cell: (319) 432-1530 Fax: (319) 221-8767 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Histogel
Thank you, John! For some strange reason, the URL in your message got garbled/multiplied, but I just took this part: http://www.thermo.com/eThermo/CMA/PDFs/Various/File_9759.pdf and that worked. The enrobing I do is to trap debris on the surface of a coral skeleton at the interface between apparently healthy tissue and bare skeleton in studying the tissue loss diseases of the corals. After solidifying the agarose, I decal the sample in neutral pH 10% EDTA, then cut (breadloaf-style) the agarose-enrobed tissue block into ~2-mm slices and put each in a cassette for processing. Perhaps my problem is the processing time needs to be increased to be sure to remove all the water from the agarose and infiltrate it completely. I don't know what the instructions mean by non-porous filter paper, either, has anyone figured this out? Esther From: histonet-boun...@lists.utsouthwestern.edu histonet-boun...@lists.utsouthwestern.edu on behalf of John Kiernan jkier...@uwo.ca Sent: Tuesday, January 21, 2014 8:54 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histogel The document at http://lists.utsouthwestern.edu/mailman/listinfo/histonet(http://www.thermo.com/eThermo/CMA/PDFs/Various/File_9759.pdf;http://www.thermo.com/eThermo/CMA/PDFs/Various/File_9759.pdf gives a thorough description of Histogel, and even says what it is - hydroxyethyl agarose. In the detailed instructions for various uses, the only confusing thing is the requirement for non-porous filter paper! John Kiernan Anatomy, UWO London, Canada = = = On 20/01/14, Elizabeth Chlipala l...@premierlab.com wrote: Esther I agree with Dusko, I fix before I put in histogel and again after the sample is placed in histogel, we have no formalin on our tissue processor, we start in 50% alcohol. I also process on a longer processing cycle, 1 hour per station and similar to Dusko's - denatured ethanol, xylene and paraplast and paraplast extra to embed. I've never had a problem (such as overprocessed tissue) with the histogel or the sample embedded in the histogel with the longer processing cycle. Most of the samples we process are cell blocks or tissue fragments such as micronized tissue constructs, which are like powder when we receive them. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] histonet-boun...@lists.utsouthwestern.edu] On Behalf Of dusko trajkovic Sent: Monday, January 20, 2014 12:47 PM To: Esther C Peters; jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histogel Esther, I mainly process cells, which have been spun down into a small pellet. Also mouse DRG's and other very small tissues. I would consider this delicate, so do not be afraid to use a longer processing program. Histogel/Agurose is what needs longer dehydrating steps. We do not use any substitute reagents, so in that aspect I cannot tell you how they will affect the processing. Our lab uses ethanol, xylene, and Paraplast paraffin. Try a test run and let me know if you were able to get successful results. Have a good Monday! Dusko From: Esther C Peters epete...@gmu.edu To: jennifer.arcand-john...@genzyme.com jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu; dusko trajkovic dunat...@sbcglobal.net Sent: Monday, January 20, 2014 11:15 AM Subject: RE: [Histonet] Histogel Thank you, Dusko! I have had the same problem with 1.5% agarose, and I tried starting the dehydration with 30% to 50% to 70% ethanol, and using different xylene substitutes. It appears that the variable whitening and shrinking happens after 100% reagent alcohol and in the xylene substitute (now using Richard-Allan Clear-Rite3). I've wondered if slow infiltration was the issue. I guess we'll try this longer processing, but I also work with delicate tissues that normally would be a short run (15 min in each reagent). Are your tissues thin/delicate/biopsy or cell preps or organ samples? No effect on them? Esther Esther C. Peters, Ph.D. Assistant Professor Environmental Science Policy George Mason University 4400 University Drive, MS 5F2 Fairfax, VA 22030- From: histonet-boun...@lists.utsouthwestern.edu histonet-boun...@lists.utsouthwestern.edu on behalf of dusko trajkovic dunat...@sbcglobal.net Sent: Monday, January 20, 2014 1:58 PM To: jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu
[Histonet] zebrafish embryos histology
Can anyone give me a method for zebra-fish embryo histology? The papers I've read show photos, but no description of histology in M M. I need to put several embryos in each block, and get the orientation correct, and put multiple sections on each slide, in hopes of getting one or two that are perfect. Any suggestions would be greatly appreciated. Thanks, Patty Lott UAB CMBD Core Lab 205-934-2007 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Cell Marque Mammoglobin (31A5) New Protocol?
don't use Ventana, but I use the Abcam concentrate at 1:100, citrate buffer- HIER and it works great. Usually cell marque AB are good, but had good luck with this one, maybe give it a try? Joelle Weaver MAOM, HTL (ASCP) QIHC From: mrs.abby.map...@gmail.com Date: Wed, 22 Jan 2014 08:11:11 -0600 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Marque Mammoglobin (31A5) New Protocol? Fellow histotechs, We've been running mammoglobin testing on our Ventana Benchmark XT. However, Cell Marque (our supplier of mammoglobin antibody) has changed the concentration. Our old concentration was 0.12 mg/mL, the new concentration is weaker at 0.04 mg/mL. The antibody is a rabbit monoclonal. We tried running the old protocol with the new concentration, but the results were not adequate (see below). We tried running the protocol given to us by Cell Marque, but theirs is specific towards the Benchmark UltraView and also returned inadequate results. We process our tissue from formalin to paraffin. Our lab uses the iVew DAB detection kit. Our old protocol: Deparaffinization Conditioner #1 (short 8 minute conditioning) Mild CC1 (mild 30 minutes conditioning) Standard CC1 (standard 60 minute conditioning) No enzyme 37C antibody incubation temperature No titration Antibody: Mammoglobin 31A5 at 0.12 mg/mL concentration for 32 minutes A/B Block Hematoxylin counterstain 4 minutes Blueing post counterstain 4 minutes Does anyone else use this product from Ventana/Cell Marque? Does anyone have a working protocol for a weaker antibody titration? Any help would be greatly appreciated! *Abby L. H. Maples, BS, MPH, HTL (ASCP)CM* Histotechnologist, Surgical Pathology Mercy Medical Center 701 10th Street SE Cedar Rapids, IA 52403 Lab Phone: (319) 398-6827 Cell: (319) 432-1530 Fax: (319) 221-8767 ___ Histonet mailing list Histonet@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] Cell Marque Mammoglobin (31A5) New Protocol?
also have IVD mammoglobin, clone 31A5 rabbit monoclonal, works really well for me at 1:75- if you need/prefer IVD from cancer diagnostics, the other I posted is ASR. Joelle Weaver MAOM, HTL (ASCP) QIHC From: mrs.abby.map...@gmail.com Date: Wed, 22 Jan 2014 08:11:11 -0600 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Marque Mammoglobin (31A5) New Protocol? Fellow histotechs, We've been running mammoglobin testing on our Ventana Benchmark XT. However, Cell Marque (our supplier of mammoglobin antibody) has changed the concentration. Our old concentration was 0.12 mg/mL, the new concentration is weaker at 0.04 mg/mL. The antibody is a rabbit monoclonal. We tried running the old protocol with the new concentration, but the results were not adequate (see below). We tried running the protocol given to us by Cell Marque, but theirs is specific towards the Benchmark UltraView and also returned inadequate results. We process our tissue from formalin to paraffin. Our lab uses the iVew DAB detection kit. Our old protocol: Deparaffinization Conditioner #1 (short 8 minute conditioning) Mild CC1 (mild 30 minutes conditioning) Standard CC1 (standard 60 minute conditioning) No enzyme 37C antibody incubation temperature No titration Antibody: Mammoglobin 31A5 at 0.12 mg/mL concentration for 32 minutes A/B Block Hematoxylin counterstain 4 minutes Blueing post counterstain 4 minutes Does anyone else use this product from Ventana/Cell Marque? Does anyone have a working protocol for a weaker antibody titration? Any help would be greatly appreciated! *Abby L. H. Maples, BS, MPH, HTL (ASCP)CM* Histotechnologist, Surgical Pathology Mercy Medical Center 701 10th Street SE Cedar Rapids, IA 52403 Lab Phone: (319) 398-6827 Cell: (319) 432-1530 Fax: (319) 221-8767 ___ Histonet mailing list Histonet@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] Cell Marque Mammoglobin (31A5) New Protocol?
Diagnostic Biosystems has Rabbit Monos Website below Mike Michael O. Thompson Director of Sales Diagnostic BioSystems Phone: 1-888-896-3350 Mobile: 412-860-1288 Office Fax: 412-727-6080 IHC Made Affordable www.dbiosys.com joelle weaver joellewea...@hotmail.com wrote: also have IVD mammoglobin, clone 31A5 rabbit monoclonal, works really well for me at 1:75- if you need/prefer IVD from cancer diagnostics, the other I posted is ASR. Joelle Weaver MAOM, HTL (ASCP) QIHC From: mrs.abby.map...@gmail.com Date: Wed, 22 Jan 2014 08:11:11 -0600 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Marque Mammoglobin (31A5) New Protocol? Fellow histotechs, We've been running mammoglobin testing on our Ventana Benchmark XT. However, Cell Marque (our supplier of mammoglobin antibody) has changed the concentration. Our old concentration was 0.12 mg/mL, the new concentration is weaker at 0.04 mg/mL. The antibody is a rabbit monoclonal. We tried running the old protocol with the new concentration, but the results were not adequate (see below). We tried running the protocol given to us by Cell Marque, but theirs is specific towards the Benchmark UltraView and also returned inadequate results. We process our tissue from formalin to paraffin. Our lab uses the iVew DAB detection kit. Our old protocol: Deparaffinization Conditioner #1 (short 8 minute conditioning) Mild CC1 (mild 30 minutes conditioning) Standard CC1 (standard 60 minute conditioning) No enzyme 37C antibody incubation temperature No titration Antibody: Mammoglobin 31A5 at 0.12 mg/mL concentration for 32 minutes A/B Block Hematoxylin counterstain 4 minutes Blueing post counterstain 4 minutes Does anyone else use this product from Ventana/Cell Marque? Does anyone have a working protocol for a weaker antibody titration? Any help would be greatly appreciated! *Abby L. H. Maples, BS, MPH, HTL (ASCP)CM* Histotechnologist, Surgical Pathology Mercy Medical Center 701 10th Street SE Cedar Rapids, IA 52403 Lab Phone: (319) 398-6827 Cell: (319) 432-1530 Fax: (319) 221-8767 ___ Histonet mailing list Histonet@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
RE: [Histonet] zebrafish embryos histology
Patty, I have personally never performed histology on zebra fish embryo's, but if I was to test it out on my own I might try using the JB4 Plus GMA kit. It just seems to me that paraffin might cause too much shrinkage and maybe using an MMA protocol might be too harsh on the tissue. Besides, given the hydrophilic nature of GMA it just might be the best overall solution. Best, Jack From: pl...@uab.edu To: histonet@lists.utsouthwestern.edu Date: Wed, 22 Jan 2014 15:00:40 + Subject: [Histonet] zebrafish embryos histology Can anyone give me a method for zebra-fish embryo histology? The papers I've read show photos, but no description of histology in M M. I need to put several embryos in each block, and get the orientation correct, and put multiple sections on each slide, in hopes of getting one or two that are perfect. Any suggestions would be greatly appreciated. Thanks, Patty Lott UAB CMBD Core Lab 205-934-2007 ___ Histonet mailing list Histonet@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] Cell Marque Mammoglobin (31A5) New Protocol?
Hi Abby, Since the concentration is about a third of the original, I would try programming the XT to run the primary for 2 hrs instead of 32 min. You may also need to up the secondary time if that isn't sufficient. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Abby L. Maples Sent: Wednesday, January 22, 2014 6:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Marque Mammoglobin (31A5) New Protocol? Fellow histotechs, We've been running mammoglobin testing on our Ventana Benchmark XT. However, Cell Marque (our supplier of mammoglobin antibody) has changed the concentration. Our old concentration was 0.12 mg/mL, the new concentration is weaker at 0.04 mg/mL. The antibody is a rabbit monoclonal. We tried running the old protocol with the new concentration, but the results were not adequate (see below). We tried running the protocol given to us by Cell Marque, but theirs is specific towards the Benchmark UltraView and also returned inadequate results. We process our tissue from formalin to paraffin. Our lab uses the iVew DAB detection kit. Our old protocol: Deparaffinization Conditioner #1 (short 8 minute conditioning) Mild CC1 (mild 30 minutes conditioning) Standard CC1 (standard 60 minute conditioning) No enzyme 37C antibody incubation temperature No titration Antibody: Mammoglobin 31A5 at 0.12 mg/mL concentration for 32 minutes A/B Block Hematoxylin counterstain 4 minutes Blueing post counterstain 4 minutes Does anyone else use this product from Ventana/Cell Marque? Does anyone have a working protocol for a weaker antibody titration? Any help would be greatly appreciated! *Abby L. H. Maples, BS, MPH, HTL (ASCP)CM* Histotechnologist, Surgical Pathology Mercy Medical Center 701 10th Street SE Cedar Rapids, IA 52403 Lab Phone: (319) 398-6827 Cell: (319) 432-1530 Fax: (319) 221-8767 ___ Histonet mailing list Histonet@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] zebrafish embryos histology
Patty, did some zebrafish work years ago, either pure histology or sections after we did whole mount ISH on the embryo's. As per Jack Ratliff's post, paraffin I found was really tough for orientation and for getting enough sections of such small embryo's. But would use, as Jack suggested, JB-4 Plus and would get beautiful sections, many embryo's in a block and multiple sections per slide. What I would do is under a dissecting scope would, with a VERY fine tool, push the multiple embryo's into an ordered row with similar orientation in the unpolymerized GMA block. Polymerize. If I wanted longitudinal sections, cut the block as is. If we wanted cross-sections, just gross cut the polymerized block and put it in a second GMA block of unpolymerized GMA and stand it up so the embryo's were on end and polymerize . Was every individual embryo correct? No! But enough were so got great HE sections or also seeing the ISH probe revealed at the cellular level. Ray, retired in Seattle - Original Message - From: Patricia F Lott pl...@uab.edu To: histonet@lists.utsouthwestern.edu Sent: Wednesday, January 22, 2014 7:00:40 AM Subject: [Histonet] zebrafish embryos histology Can anyone give me a method for zebra-fish embryo histology? The papers I've read show photos, but no description of histology in M M. I need to put several embryos in each block, and get the orientation correct, and put multiple sections on each slide, in hopes of getting one or two that are perfect. Any suggestions would be greatly appreciated. Thanks, Patty Lott UAB CMBD Core Lab 205-934-2007 ___ Histonet mailing list Histonet@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] Re: zebrafish embryos histology
Hi Patty, We did some of these in paraffin in my other life in Kansas City. Essentially for these we processed them in paraffin, and would embed them in Ls, letting gravity do what it does best. We would line them up side by side, as perpendicular to the edge as we could. We had a stereomicroscope attached to an arm so we could see what we were doing with the small samples, and would use either a toothpick, wooden stick, or very fine brush to orient them in the embedding L. If we needed transverse sections, we would just mount the block as such in a standard specimen clamp and section. Honestly for us it was much quicker than using plastics, and it gave us the ability to do regular specimen and IHC stains on them if needed. Teri Johnson Manager, Histology Genomics Institute for Novartis Research Foundation San Diego, CA 858-332-4752 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cell Marque Mammoglobin (31A5) New Protocol?
Thank you everyone! You've all been so helpful regarding the mammaglobin. Ventana has called me and I think we got things figured out (at least tenatively) and the wheels are in motion. If it still doesn't work, our rep will come out and see us. Thanks again for your input! Great ideas abound!! On Wed, Jan 22, 2014 at 10:44 AM, Thomas Jasper thomas.jas...@deaconess.com wrote: Hi Abby, I have a couple of suggestions - 1) Try to extend your incubation times. You may need to do a couple-three of these to get a decent result. 2) Contact Ventana and have a field specialist come out and help you with working out this problem on the Benchmark. My experiences with Ventana have been positive in this regard and I've always found them willing to help in any way possible. You may have already thought of, and done these things, I don't know. Just thought I'd mention them. Good luck to you, Tom Jasper Thomas Jasper HT (ASCP) BAS AP/CP Supervisor Deaconess Hospital 600 Mary Street Evansville, IN 47747 thomas.jas...@deaconess.com 812-450-2485 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto: histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Abby L. Maples Sent: Wednesday, January 22, 2014 8:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Marque Mammoglobin (31A5) New Protocol? Fellow histotechs, We've been running mammoglobin testing on our Ventana Benchmark XT. However, Cell Marque (our supplier of mammoglobin antibody) has changed the concentration. Our old concentration was 0.12 mg/mL, the new concentration is weaker at 0.04 mg/mL. The antibody is a rabbit monoclonal. We tried running the old protocol with the new concentration, but the results were not adequate (see below). We tried running the protocol given to us by Cell Marque, but theirs is specific towards the Benchmark UltraView and also returned inadequate results. We process our tissue from formalin to paraffin. Our lab uses the iVew DAB detection kit. Our old protocol: Deparaffinization Conditioner #1 (short 8 minute conditioning) Mild CC1 (mild 30 minutes conditioning) Standard CC1 (standard 60 minute conditioning) No enzyme 37C antibody incubation temperature No titration Antibody: Mammoglobin 31A5 at 0.12 mg/mL concentration for 32 minutes A/B Block Hematoxylin counterstain 4 minutes Blueing post counterstain 4 minutes Does anyone else use this product from Ventana/Cell Marque? Does anyone have a working protocol for a weaker antibody titration? Any help would be greatly appreciated! *Abby L. H. Maples, BS, MPH, HTL (ASCP)CM* Histotechnologist, Surgical Pathology Mercy Medical Center 701 10th Street SE Cedar Rapids, IA 52403 Lab Phone: (319) 398-6827 Cell: (319) 432-1530 Fax: (319) 221-8767 ___ Histonet mailing list Histonet@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] zebrafish embryos histology
Hi Patty, You sort of piqued my curiosity of what is going on in the zebra fish world and found this youtube, 6 minute film http://www.youtube.com/watch?v=kZDwo20hl1Efeature=youtu.be About what is going on at Welcome Trust Research on zebra fish but maybe you know all this already. Anyway, I think the film is pretty neat with some incredible (not histology) but CONFOCAL slices through the fish. We had success doing this by confocal on whole-mount but again when studying gene manipulation and cell signal distribution and wanting histology in embryo's, had better luck with GMA. Paraffin as mentioned can certainly be done and is certainly easier but we could never get the resolution or number of sections we desired using paraffin. Maybe the people in lab from this film can give you some suggestions. Ray, retired in Seattle - Original Message - From: koelli...@comcast.net To: Patricia F Lott pl...@uab.edu Cc: histonet@lists.utsouthwestern.edu Sent: Wednesday, January 22, 2014 8:02:24 AM Subject: Re: [Histonet] zebrafish embryos histology Patty, did some zebrafish work years ago, either pure histology or sections after we did whole mount ISH on the embryo's. As per Jack Ratliff's post, paraffin I found was really tough for orientation and for getting enough sections of such small embryo's. But would use, as Jack suggested, JB-4 Plus and would get beautiful sections, many embryo's in a block and multiple sections per slide. What I would do is under a dissecting scope would, with a VERY fine tool, push the multiple embryo's into an ordered row with similar orientation in the unpolymerized GMA block. Polymerize. If I wanted longitudinal sections, cut the block as is. If we wanted cross-sections, just gross cut the polymerized block and put it in a second GMA block of unpolymerized GMA and stand it up so the embryo's were on end and polymerize . Was every individual embryo correct? No! But enough were so got great HE sections or also seeing the ISH probe revealed at the cellular level. Ray, retired in Seattle - Original Message - From: Patricia F Lott pl...@uab.edu To: histonet@lists.utsouthwestern.edu Sent: Wednesday, January 22, 2014 7:00:40 AM Subject: [Histonet] zebrafish embryos histology Can anyone give me a method for zebra-fish embryo histology? The papers I've read show photos, but no description of histology in M M. I need to put several embryos in each block, and get the orientation correct, and put multiple sections on each slide, in hopes of getting one or two that are perfect. Any suggestions would be greatly appreciated. Thanks, Patty Lott UAB CMBD Core Lab 205-934-2007 ___ Histonet mailing list Histonet@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] RE: Zebra-fish embryo histology
Patty, Our lab has processed zebra fish embryos for paraffin sectioning. The embryos were fixed overnight in 4% paraformaldehyde and processed on a biopsy program without any heat. The key is to pop the swim bladders of the fish so that they will lay flat during embedding. We put multiple pieces into each biopsy sized mold and were able to get a good section with intestine on at least one of the fish in each block. It is imperative that you soak the blocks, on a paper towel moistened with diluted ammonium hydroxide, on ice for at least an hour to get decent sections because the tissue is very dry. Best Regards, Bonnie Gambichler, HT ( ASCP ) JHU Oncology Tissue Services Lab ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Retaining unstained glass slides
How long due most labs retain glass slides? Renee H. Workman Histology Supervisor Virginia Urology 9105 Stony Point Drive Richmond, VA 23235 W: 804-527-1316 | F: 804-270-0917 rhwork...@uro.commailto:rhwork...@uro.com | www.uro.comhttp://www.uro.com Disclaimer: The email and 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 original recipient or the person responsible for the delivering the email to the intended recipient, be advised that you have received this email in error, and that any use, dissemination, forwarding, printing or copying of this email is strictly prohibited. If you received this email in error, please delete it from your system without copying it, and notify the sender by reply email so that our address record can be corrected. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Retaining unstained glass slides
Follow the guidelines of your accrediting agency. CAP = 10 years minimum ANP.12500 Record Retention Phase II Surgical pathology records and materials are retained for an appropriate period. NOTE 1: Minimum requirements for surgical pathology, providing these are not less stringent than local, state and national regulations, are: 1. Accession log records - 2 years 2. Wet tissue (stock bottle) - 2 weeks after final report 3. Paraffin blocks - 10 years (subject to Note 2, below) 4. Glass slides (including control slides) and reports - 10 years (slides must remain readable for this period) 5. Surgical pathology reports - 10 years (see Notes 3 and 4, below) 6. Fluorochrome-stained slides - at the discretion of the laboratory director 7. Fine needle aspiration slides - 10 years 8. Images of FISH studies - 10 years (see Note 5, below) Walter Benton HT(ASCP)QIHC Histology Supervisor Chesapeake Urology Associates 806 Landmark Drive, Suite 127 Glen Burnie, MD 21061 443-471-5850 (Direct) 410-768-5961 (Lab) 410-768-5965 (Fax) ChesapeakeUrology.com Voted a Best Place to Work by Baltimore and Modern Healthcare Magazines. From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Renee H. Workman [rhwork...@uro.com] Sent: Wednesday, January 22, 2014 1:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Retaining unstained glass slides How long due most labs retain glass slides? Renee H. Workman Histology Supervisor Virginia Urology 9105 Stony Point Drive Richmond, VA 23235 W: 804-527-1316 | F: 804-270-0917 rhwork...@uro.commailto:rhwork...@uro.com | www.uro.comhttp://www.uro.com Disclaimer: The email and 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 original recipient or the person responsible for the delivering the email to the intended recipient, be advised that you have received this email in error, and that any use, dissemination, forwarding, printing or copying of this email is strictly prohibited. If you received this email in error, please delete it from your system without copying it, and notify the sender by reply email so that our address record can be corrected. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: The information contained in this electronic message is intended solely for the personal and confidential use of the designated recipient(s) named above and may contain information that is protected from disclosure under applicable law. If you are not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this transmission is strictly prohibited. If you have received this transmission in error, please notify the transmitting person/department immediately by email or telephone (410) 581-5881 and delete the message without making a copy. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] lyophilizing company
Histonetters: My lab is needing to lyophilize some skin tissue. Is there a company that offers such services? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Buffer pH for IHC
What does everyone use to test the pH of their buffers for IHC? Are pH strips adequate or do you have to have an actual pH meter? If you use a meter, any suggestions on a make and model? Laurie Colbert, HT (ASCP) Histology Supervisor PATH MD 8158 Beverly Blvd. Los Angeles, CA 90048 (323) 648-3214 direct (424) 245-7284 main lab The information contained in this transmission may contain privileged and confidential information, including patient information protected by federal and state privacy laws. It is intended only for the use of the person(s) named above. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution, or duplication of this communication is strictly prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Buffer pH for IHC
pH meter; Corning. On Wed, Jan 22, 2014 at 3:21 PM, Laurie Colbert lcolb...@pathmdlabs.comwrote: What does everyone use to test the pH of their buffers for IHC? Are pH strips adequate or do you have to have an actual pH meter? If you use a meter, any suggestions on a make and model? Laurie Colbert, HT (ASCP) Histology Supervisor PATH MD 8158 Beverly Blvd. Los Angeles, CA 90048 (323) 648-3214 direct (424) 245-7284 main lab The information contained in this transmission may contain privileged and confidential information, including patient information protected by federal and state privacy laws. It is intended only for the use of the person(s) named above. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution, or duplication of this communication is strictly prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Jan Shivers Senior Scientist IHC/Histology Section Head Pathology Teaching Program Veterinary Diagnostic Laboratory University of Minnesota 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive...@umn.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] How to do validation of PLA2R antibody on serum
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