[Histonet] thanks
Many thanks to all who contribute and keep the histonet running. I'm working my last day here at USDA-ARS ADRU within WSU which has been a very rewarding and enjoyable occupation. I was honored to be on 14 publications in 12 years with a few more in the works. Due to Federal budget constraints, they don't plan on replacing me with another BS An. Sci. ASCP HT. Happy New Year to you all! Thanks again Tom T= Thomas C. Truscott ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: lab math: ihc dilution
When I solve your equation V1 X 1560 ug/ml = 10ml X 0.8 ug/ml I get .005 ml. Therefore the dilution would be closer to 1:2000. I would usually divide the 1560 ug/ml by 0.8 ug/ml and get 1950 therefore 1:1950 would be more accurate . Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kathleen Jones Sent: Friday, November 21, 2014 11:50 AM To: Sheryl Stephenson; histonet@lists.utsouthwestern.edu; Elizabeth Chlipala Subject: [Histonet] RE: lab math: ihc dilution Hi Liz, Wouldn't you dilute in 999.5ul of diluent, not .5. 1ml=1000ul Otherwise I got the same answer... Kathleen Kathleen Jones Research Technologist Pathology/Microbiology AVC - UPEI (902)213-2207 Elizabeth Chlipala l...@premierlab.com 21/11/2014 3:35 PM Sheryl Here is how I would approach the problem. We would use the basic mathematical equation of the following (in order for this to work you need to keep the units the same) V1 x C1 = V2 x C2 V1 - volume of stock solution needed - this is what you are solving for C1 - concentration of stock solution - you know this its 1.56 mg/ml or 1560 ug/ml (there are 1000 ug in one mg) V2 - volume of final solution needed - I put an arbitrary volume of 10 mls needed C2 - concentration of final solution - you know this its 0.8ug/ml V1 x 1560 ug/ml = 10ml x 0.8 ug/ml --- see how I made sure the units were the same I changed the mg/ml to ug/ml V1 = .0005 mls and then I would need 9.9995 mls of diluent to make up my 0.8ug/ml concentration, you can change this to be expressed in microliters by multiplying each number by 1000. So in uls - V1 = 0.5 uls and .5 uls from there you can determine the titer which is your total volume divided by the volume of concentrated antibody 10/.0005 That would be a 1:20,000 dilution For a dilution such as this I would make up a stock solution of either 1:100 or 1:1000 and then dilute from there. Now its time for the rest of you to check my math, did I do this correctly? I went over it a few times, but you never know.. Have a GREAT Weekend! 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 March 10, 2014 is Histotechnology Professionals Day 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] On Behalf Of Stephenson, Sheryl Sent: Friday, November 21, 2014 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lab math: ihc dilution To all you math whiz out there, please help with this math dilution. If my Ab conc is 1.56 mg/mL and they want to optimize to 0.8 ug/mL, what dilution should I use? Thanks, Sheryl ___ Histonet mailing list Histonet@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] RE: lab math: ihc dilution
You are so right-that's why I always try to double check everything even my spellink. Tom T -Original Message- From: Elizabeth Chlipala [mailto:l...@premierlab.com] Sent: Friday, November 21, 2014 12:05 PM To: Truscott, Tom; 'histonet@lists.utsouthwestern.edu' (histonet@lists.utsouthwestern.edu) Subject: RE: [Histonet] RE: lab math: ihc dilution You are right I got it wrong, see no matter how many time you run the math, you still come up with problems 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 March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: Truscott, Tom [mailto:ttrus...@vetmed.wsu.edu] Sent: Friday, November 21, 2014 1:01 PM To: Kathleen Jones; Sheryl Stephenson; histonet@lists.utsouthwestern.edu; Elizabeth Chlipala Subject: RE: [Histonet] RE: lab math: ihc dilution When I solve your equation V1 X 1560 ug/ml = 10ml X 0.8 ug/ml I get .005 ml. Therefore the dilution would be closer to 1:2000. I would usually divide the 1560 ug/ml by 0.8 ug/ml and get 1950 therefore 1:1950 would be more accurate . Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kathleen Jones Sent: Friday, November 21, 2014 11:50 AM To: Sheryl Stephenson; histonet@lists.utsouthwestern.edu; Elizabeth Chlipala Subject: [Histonet] RE: lab math: ihc dilution Hi Liz, Wouldn't you dilute in 999.5ul of diluent, not .5. 1ml=1000ul Otherwise I got the same answer... Kathleen Kathleen Jones Research Technologist Pathology/Microbiology AVC - UPEI (902)213-2207 Elizabeth Chlipala l...@premierlab.com 21/11/2014 3:35 PM Sheryl Here is how I would approach the problem. We would use the basic mathematical equation of the following (in order for this to work you need to keep the units the same) V1 x C1 = V2 x C2 V1 - volume of stock solution needed - this is what you are solving for C1 - concentration of stock solution - you know this its 1.56 mg/ml or 1560 ug/ml (there are 1000 ug in one mg) V2 - volume of final solution needed - I put an arbitrary volume of 10 mls needed C2 - concentration of final solution - you know this its 0.8ug/ml V1 x 1560 ug/ml = 10ml x 0.8 ug/ml --- see how I made sure the units were the same I changed the mg/ml to ug/ml V1 = .0005 mls and then I would need 9.9995 mls of diluent to make up my 0.8ug/ml concentration, you can change this to be expressed in microliters by multiplying each number by 1000. So in uls - V1 = 0.5 uls and .5 uls from there you can determine the titer which is your total volume divided by the volume of concentrated antibody 10/.0005 That would be a 1:20,000 dilution For a dilution such as this I would make up a stock solution of either 1:100 or 1:1000 and then dilute from there. Now its time for the rest of you to check my math, did I do this correctly? I went over it a few times, but you never know.. Have a GREAT Weekend! 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 March 10, 2014 is Histotechnology Professionals Day 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] On Behalf Of Stephenson, Sheryl Sent: Friday, November 21, 2014 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lab math: ihc dilution To all you math whiz out there, please help with this math dilution. If my Ab conc is 1.56 mg/mL and they want to optimize to 0.8 ug/mL, what dilution should I use? Thanks, Sheryl ___ Histonet mailing list Histonet@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] Jores, Klotz and that pesky chloral hydrate
Hi Molly, You might just want to fix in buffered 10% formalin and then rinse specimens well in water ( maybe 20 minutes) before using in class and then put back in formalin. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Molly Murphy Sent: Thursday, November 20, 2014 1:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Jores, Klotz and that pesky chloral hydrate Hi All, I am looking for a tissue fixative to preserve gross specimens for a veterinary pathology teaching lab (eg. no histo, only gross specimens). I have used Jore's in the past, and Klotz has been recommended, but the chloral hydrate is a problem child due to its status as a controlled substance (eg disposal is a hassle). Does anyone have any thoughts about just leaving the chloral hydrate out? Or, an alternative fixative that doesn't have the chloral hydrate? I have access to a refrigerator for samples, and would *hopefully* keep the specimens for a year or two. Thanks a bunch, M -- Molly Murphy DVM, Ph.D Assistant Professor of Veterinary Pathology College of Natural Sciences Mathematics University of Alaska Fairbanks Office: (907) 474-1990 Fax: (907) 474-1932 ___ Histonet mailing list Histonet@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: processing and staining preserved specimen
Hi Adelle, Biopsy sizes will matter, but you might shorten some of these times, but be sure to add at least one more Histoclear to make sure there is no alcohol in it and at least one more paraffin to make sure there is no histoclear carryover.If you go back to 70% at any time you will have to start over, so try to get a janitor or someone to start early or a coach to finish late. Make sure you keep things safe, because of all the flammables. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Schade, Adelle Sent: Friday, October 17, 2014 10:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing and staining preserved specimen Hello everyone, I have been following and learning from this listserv for the past two years, I have enjoyed all of the advice this forum presents. This is my first inquiry for the group. I am a high school Anatomy and Physiology teacher and a grad student at Thomas Jefferson (Philadelphia, PA). In our research lab, we process, section, embed and image for all research projects. I was lucky enough to acquire some used histology equipment for my high school lab this school year. I am teaching the students about histology and the processing. We have an embedding station, microtome, staining dishes and a good microscope to image. We do not have an automatic tissue processor so we are processing manually (I prepare all solutions for the students). We are taking biopsy samples from our preserved mink that we are dissecting at the moment. I have processed the tissue using the following method and have pretty good results, not great. (we are doing a basic HE stain to see the cells of the tissue). If we do not complete the manual processing method and need to leave school for the day, I move the cassettes into 70% EtOH until the next morning when we continue the process. I am wondering if anyone has manually processed preserved specimen tissue in the past and has any advice/ protocol that you would suggest? This is what I am using: Processing Method 1. 70% EtOH: 1 hour 2. 80% EtOH: 1 hour 3. 90% EtOH: 1 hour 4. 95% EtOH: 2 hours 5. 100% EtOH: 2 hours 6. Histoclear II: 2 hours 7. Paraffin: 2 hours Also- I will eventually be looking for grants to try to purchase an automatic tissue processor- this would alleviate the time issues we are facing. Does anyone know of grants that would focus on the educational aspect of histology? Thank you for your time and have a great weekend! Adelle Schade Graduate student: Jefferson School of Biomedical Science Ms. Adelle L. Schade, B.S., M.Ed. Anatomy and Physiology Conrad Weiser High School 44 Big Spring Rd. Robesonia, PA 19551 a_sch...@conradweiser.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: rolling sections
Hi Roberta, Check with those requesting to see if you can cut the equilivant but with thicker sections that will roll- like 5 20's or 2 50's. The rolls are lots easier to get into the tubes. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pam Marcum Sent: Thursday, June 12, 2014 8:00 AM To: Helen Fedor Cc: Histonet (histonet@lists.utsouthwestern.edu); Roberta Horner Subject: Re: [Histonet] RE: rolling sections That is how we do it also. Since they are not really interested in the sections being flat we just pick them and put them directly into the tube.If I am really having issues I will actually warm the block with my finger to help it roll. If they are doing DNA or RNAase ou may need to wear glove to prevent any contamination of the blocks. Pam Marcum UAMS - Original Message - From: Helen Fedor hfe...@jhmi.edu To: Roberta Horner r...@psu.edu, Histonet (histonet@lists.utsouthwestern.edu) histonet@lists.utsouthwestern.edu Sent: Thursday, June 12, 2014 9:54:45 AM Subject: [Histonet] RE: rolling sections Hi, I think that it is not necessary to actually get them to roll. We just collect all of the sections and put them into the tube. Scrunched, not rolled. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Roberta Horner Sent: Thursday, June 12, 2014 10:42 AM To: Histonet (histonet@lists.utsouthwestern.edu) Subject: [Histonet] rolling sections I have some researchers that want to do PCR. They want 10 - 10u sections in a micro-centrifuge tube. The only way to get the sections in the tube is for the sections to roll. How do you get sections to roll when you want them to roll? I've tried room temperature, on ice, brand new sharp blade, dull blade and I can still get some really nice ribbons. When I want a thick ribbon it will roll, darn that Murphy and his laws. Roberta Horner Animal Diagnostic Lab Penn State University ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Transporting blocks/slides between sites
Hi Sharon, If you are trying to be secure enough against tampering that it would stand up in court- the zip tie method probably isn't enough. I would try to get a used small set of the larger locking safe deposit boxes or maybe a group security box from grainger. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Morken, Timothy Sent: Monday, April 07, 2014 3:06 PM To: 'Sharon Scalise' Cc: Histonet Subject: [Histonet] RE: Transporting blocks/slides between sites Sharon, We transport slide flats and tissue containers/cassettes baskets for processing (most grossing at one site, all tissue processing at another site). We use Colman 50QT coolers with handles and wheels to transport. They take a beating. We have about 10 of these because we have them receiving material at each of 3 sites and then four being transported at all times (30 min pickup schedule, each direction). It is a cheap and effective solution. We use half for paper/slide only (clean set) and half for tissue samples. If you have less material, then maybe a smaller version of these would work for you. Tim Morken Supervisor, Histology, Electron Microscopy and Neuromuscular Special Studies UC San Francisco Medical Center San Francisco, CA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sharon Scalise Sent: Monday, April 07, 2014 2:53 PM To: 'histonet' (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Transporting blocks/slides between sites We are looking for a better way to transport blocks and slides between our hospital sites. We want the system to be secure so that as we track from site to site we can be assured that the contents have not been tampered with. We currently are using plastic totes that have lids the can be secured using a zip tie. The problem is that during transport (over time) these totes get pretty beat up and end up with cracks and holes in them. We feel there is probably a better system so I am looking to the experience of the histonet users for suggestions. Thank you! Sharon Scalise, HTL(ASCP) Histology Supervisor-Anatomic Pathology Beaumont Health System 3601 W. 13 Mile Rd. Royal Oak, MI 48073 248 898-5981 sscal...@beaumont.edumailto:sscal...@beaumont.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
[Histonet] RE: Clinical histology to Research histology
Hi Cassie, For me, it has been mostly positive changes. I went from feeling like a robot cranking out slides and rarely hearing about any impact on the patient to being involved from start to finish on published articles in research journals. I've been able to use cutting edge technology to help develop new diagnostic tests. For the most part I've been able to work independently, at my own pace. The annual budget crunch can be annoying, but good research usually gets funded. I've had to cut way more serial sections than I did in clinical and do primarily IHC and work closely with WADDL for processing and a few special stains. Sometimes, you have to know when to quit trying to make something work. I could probably ramble on. Any specific questions? Good luck with your endeavors! Tom Truscott -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Davis, Cassie Sent: Tuesday, February 04, 2014 6:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Clinical histology to Research histology Hello Histo World, please share your experience from going from clinical histology to research histology...What are the major difference? Are there complications or pleasant surprises? Cassandra Davis cda...@che-east.org 302-575-8095 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Fungus contamination
Hi Judith, If you have made completely fresh solutions, even any stock reagents and still have the problem, then the fungus may be arriving from another source. I once had a problem of pollen landing on my water bath and showing up on the stains that would detect it. You may have fungus on other slides but only showing up in the PAS and GMS. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pardue, Judith Sent: Tuesday, February 04, 2014 7:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fungus contamination Having a problem with fungus on top of both the pas and gms stain. There are no solutions used on both test. Other than distilled water we can not figure it out. All solutions have been changed and clean containers used. Judith Gale Pardue Histology Supervisor judith_pardue@memorial .org This electronic mail and any attached documents are intended solely for the named addressee(s) and contain confidential information. If you are not an addressee, or responsible for delivering this email to an addressee, you have received this email in error and are notified that reading, copying, or disclosing this email is prohibited. If you received this email in error, immediately reply to the sender and delete the message completely from your computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] marking tiny specimens
Hi Cheryl, If you can, use more eggs. Then process them into a pellet with histogel. Upon sectioning, many different cross-sections will be visible in the same section. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cheryl Crowder Sent: Thursday, January 30, 2014 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] marking tiny specimens I am processing some extremely small specimens - pin tip size. These are eggs with a protein covering. I have tried using eosin to color the tissues before processing but the color came out before paraffin. The coating on the eggs will not absorb the dye. Does anyone have a suggestion for dyeing or marking these tissues so I can see them better to embed. Thanks in advance, Cheryl Cheryl Crowder, BA, HTL(ASCP) ___ Histonet mailing list Histonet@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] Freezer for tissue storage
We have both upright and chest and both have their pros and cons. We have had trouble with electrical circuits which aren't matched to the load that these freezers can put on them. An electrician told us that it can put an tremendous strain on the freezer motor if the circuit isn't right and two or three freezers start up at the same time- leading to shortened life of the freezer. So- make sure a good electrician is involved in your freezer set-up. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Hugh Luk Sent: Friday, December 13, 2013 1:41 PM To: histonet@lists.utsouthwestern.edu; richard.car...@hhchealth.org Subject: [Histonet] Freezer for tissue storage Dr. Cartun, Agreed, chest freezers have less frost/door problems/sealing issues and seem to live longer. Stand-ups maximize footprint/space efficiency and allow for easier, horizontal-rack pulling for archiving and inventory. Due to volume and space constraints, the UH cancer center buys stand-ups. We have over a hundred. My personal caution is for the newer Thermo-Revco freezers, especially the ultra-low Thermo UXF in mostly 24.1 cu. ft.. Perhaps this is just our bad luck but an abnormal number failed (7/9), and three were within the first year. The unfortunate units were purchased over the last four years. Our bio-engineer has asked us to avoid buying Revco (after endorsing Revco/Harris for many years) and buy New Brunswick/Eppendorf or Sanyo ULT freezers, for the immediate future. We're not endorsing Sanyo or New Brunswick, we're just staying away from a brand that has recently given us trouble. Hopefully, it is different in your area, as Hawaii has logistical issues that may have exacerbated this issue (maybe it was that looong boat trip)? Just my $0.02. (And $0.02 will not buy anything, anymore.) Hugh UHCC path shared resource manager Honolulu, HI -- Date: Thu, 12 Dec 2013 18:51:57 + From: Cartun, Richard richard.car...@hhchealth.org Subject: [Histonet] Freezer for tissue storage To: Histonet histonet@lists.utsouthwestern.edu Message-ID: 9215bd4b0ba1b44d962a71c758b68d2e018a0...@hhcexchmb05.hhcsystem.org Content-Type: text/plain; charset=iso-8859-1 I need a recommendation for a -80 degree C. freezer for storing tissue specimens. Do you prefer upright vs. chest? Thanks. 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 richard.car...@hhchealth.orgmailto:richard.car...@hhchealth.org 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, or an employee or agent responsible for delivering the message to the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message, including 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
RE: [Histonet] Rodent eye paraffin microtomy
Hi Jackie, I haven't done a lot of eyes, but mainly sheep lenses. I seemed to have more trouble with crushing artifact when trimming in- so I tried not to trim too close and do the closer trimming on the microtome- I also trimmed before they got too hard from fixing. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bea DeBrosse-Serra Sent: Thursday, December 12, 2013 9:13 AM To: 'b427...@aol.com'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Rodent eye paraffin microtomy Let the eye soak in ice water before cutting for a very long time. Bea Beatrice DeBrosse-Serra HT(ASCP)QIHC Isis Pharmaceuticals Antisense Drug Discovery 2855 Gazelle Ct. Carlsbad, CA 92010 760-603-2371 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of b427...@aol.com Sent: Thursday, December 12, 2013 8:48 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Rodent eye paraffin microtomy I have a study coming up where obtaining perfect lens histology is critical. We do a pretty good job routinely, but since the lens is of interest, I would appreciate any tricks and techniques that can help us improve our paraffin lens histology within intact rodent eyes. Thanks, Jackie O' ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] (no subject)
Hi Cherie, Since the top section is consistently better, then the possibilities lie in what happens after they are on the slide. As Toni recommends slides that are heated or deparaffinized vertically may have left over paraffin on the lower regions.Maybe you could also try depar with them up on their long edges. Staining slides vertically might also give this appearance if reagent times and rinsing are too short. Staining slides flat would point to slides not level or reagents not evenly dispersed on the slide. I would not rule out thick-thin on ribbons but that is probably not the case since the good section is always at the top. Good luck, Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rathborne, Toni Sent: Wednesday, December 04, 2013 10:17 AM To: 'Chapman, Cherie J.'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] (no subject) Could it be the heating/deparaffinization process? If the upper sections are staining more evenly, then maybe they are free from residual paraffin. Try extending the time in the ovens and/or xylene. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Chapman, Cherie J. Sent: Wednesday, December 04, 2013 1:12 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hello all, I am looking for suggestions on issues with our HE stain. I supervised a Veterinary Diagnostic lab for over 27 years and produced top quality sections, HE's, special stains and IHC on a variety of different species in our lab. I am currently working in a Dermatopathology Lab and I am finding inconsistent staining with our HE's. Working with just skin is a challenge all on its own. We have made changes to our staining protocol and just not happy with the end product. What we are observing is inconsistent staining on levels on the same slide. The top section seems to stain more evenly than the middle and bottom sections. I can actually see three different shades of color. The specimens are ribboned sections so I know it is not from thick and thin sections. We have looked at our processing times, microwave vs. oven times, staining reagents, different brands of hematoxylin and eosin, adjustments on staining times, tap water compared to distilled water. Our main processor is the Thermo Scientific STP-420 and our back up is the Sakura VIP V processor.I have been working with Thermo technical support thinking it might be a processing issue. We have a Leica ST5020 Multstainer/CV5030 Robotic Cover slipper we have made several changes that the technical teams has suggested to the reagents and staining time. It's still not the quality that we are looking for. I have had culligan techs out several times to see if it could be something with the water. We can run 100 slides the same day, same reagents and protocol and the HE color is so inconsistent. I would appreciate any suggestions in this matter. Cherie Chapman, BS, HT, HTL (ASCP) Associate Director of Dermatopathology Laboratory University of Missouri Department of Dermatology University Physicians Medical Building Phone: (573) 884-0123 Fax: (573) 884-0834 ___ Histonet mailing list Histonet@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] Qdots
If anyone has had success with solving background issues with Ventana's Q-Dot labeling, would you please contact me? Thankyou, Tom Truscott ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Processing thin slices of mouse cerebellum
Hi Kathleen, Several years and a couple employers ago we attached small biopsies (for proper orientation) to small squares of cucumber (which had been dehydrated in several changes of alcohol) with a egg-albumin/glycerol mixture. The bx/cucumber unit was embedded together after processing and cut and stained as a unit with no adverse effects. Tom Truscott -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of kgrob...@rci.rutgers.edu Sent: Wednesday, May 29, 2013 11:14 AM To: histonet Subject: [Histonet] Processing thin slices of mouse cerebellum To all, We have these very thin slices of mouse cerebellum that need to be processed into paraffin. The problem is that they need to be kept flat. We have tried sponges, but some of the slices are so small that they shrink (processed with our biopsy program, of course; if you want details, let me know) and get lost in the holes of the sponges. We have biopsy cassettes, but the slices will still tumble and not stay flat. We have processed other things in lens paper, but I had to scrape the tissue with the paraffin into the mold. I have not tried Histogel, though-maybe embed the slices flat in that first and then process it? Is there anything else? Thanks so much! Kathleen Principal Lab Technician Neurotoxicology Labs Molecular Pathology Facility Core Dept of Pharmacology Toxicology Rutgers, the State University of NJ 41 B Gordon Road Piscataway, NJ 08854 (848) 445-1443 FAX (732) 445-6905 ___ Histonet mailing list Histonet@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] H pylori and Ventana instruments
I haven't used HP, but have noticed this stippled effect especially when using 40X on the microscope and looking at slides labeled with an Ventana alk-phos kit. I seem to have solved the problem by making sure no trace of H2O is left on the slides. It helps after washing slides to run them thru a few changes of alcohol then one acetone and then a couple xylene before coverslipping. Ventana suggested the acetone step. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Roger Heyna Sent: Wednesday, May 08, 2013 8:41 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H pylori and Ventana instruments We use Ventana's HP antibody, and we do see some of the small, non-specific globs of stain, but the pathologists are comfortable differentiating between the true and false staining since they are familiar with this artefact. Roger Heyna Rae Staskiewicz raest...@grics.net 5/7/2013 12:56 PM Hi all, Those of you using Ventana instrumentation, could you tell me what H pylori antibody you are using, and whether or not you are experiencing non-specific staining, or staining that looks specific but with a stippled effect. Thanks Rae Staskiewicz UnityPoint Health-Methodist ___ Histonet mailing list Histonet@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] Recycled Xylene on Tissue Processor
If the xylene was not recycled properly and retained too much alcohol, then the extra alcohol could be the culprit and do the drying'. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Wednesday, January 23, 2013 12:47 PM To: Laurie Colbert; Histonet Post (histonet@lists.utsouthwestern.edu) Subject: Re: [Histonet] Recycled Xylene on Tissue Processor If xylene is recycled properly and has no alcohol residues, i should not pose any problems. It will clear just like the original pure xylene, at least that is what I found with my recycled xylene for more than 10 years. The problem you describe should have a different cause. René J. From: Laurie Colbert lcolb...@pathmdlabs.com To: Histonet Post (histonet@lists.utsouthwestern.edu) histonet@lists.utsouthwestern.edu Sent: Wednesday, January 23, 2013 1:55 PM Subject: [Histonet] Recycled Xylene on Tissue Processor Has anyone that uses recycled xylene on the tissue processor ever noticed that it dries out the biopsies (specifically GI bx's)?? We are having issues with the GI's being dried out, and I'm wondering if this may be the cause. Laurie Colbert ___ Histonet mailing list Histonet@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] annoying crystals on sections
Hi Wayne, I had lots of problems with round irregular crystals, but have greatly improved my slides by limiting baking at 57 degrees to 1/2 hour. We use a paraffin with plastic in the mix, and I think the plastic globs up under some conditions, like no or not enough xylene to dissolve it out. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of E. Wayne Johnson Sent: Tuesday, August 21, 2012 8:09 AM To: Debra Siena; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] annoying crystals on sections we are using a Sakura DRS2000 and we are 3 x 3 minutes in xylene and we have been keeping it fresh. The staining is good now but we still see the crystals. If it were paraffin we should see unstained spots on the slide I think. I have gone to an aqueous 1% HCl today after hematoxylin for regression and that seems to be cleaning them up on most of the slides. I cleaned out some of the plumbing and cleaned some calcium out of the pipes. We are using Harris hematoxylin that we purchase. We have a tried different counterstains but it seems to make no difference. We are using a Sakura tissue processor for overnight processing of cassettes. the embedding is going good and we get nice flat thin sections. We are fixing tissues with neutral phosphate buffered formalin but still see some formalin pigment. We are cleaning that up with picric acid in etoh. We find we still need that and the formalin pigment is brown to dark brown. These problem crystals are round irregular to rhomboidal some times sort of large and flat about the size of a cell and they are clear. I thought they were formalin pigment at first and fiddled with the Picric Acid, and tried Ammonia in Alcohol to get rid of formalin pigment and finally decided that it was not formalin pigment. I thought it might be something from Scott's tap water (Mg++) so i dropped that and tried bluing with NH4+ and it didnt help any. I tried blueing just with tap water. Nice result but still the crystals. The aqueous HCl seems to be working and is not harming the nuclei so I may have a sort of solution and am calling it calcium crystals in the water until I know better. I may look for some sort of filter to put in the water line. E Wayne Johnson DVM Enruikang Ag Tech MOA Feed Industry Centre China Agriculture University Beijing On 8/21/2012 7:51 PM, Debra Siena wrote: could it be paraffin? Debbie Siena HT(ASCP)QIHC Technical Manager | StatLab Medical Products 407 Interchange St. | McKinney, TX 75071 Direct: 972-436-1010 x229 | Fax: 972-436-1369 dsi...@statlab.com | www.statlab.com - Original Message - From: e...@pigsqq.org [mailto:e...@pigsqq.org] Sent: Tuesday, August 21, 2012 05:46 AM To: Debra Siena Subject: Re: [Histonet] annoying crystals on sections We just now ran the statlab version protocol from StatLab's website, using an alcoholic eosin. No doubt that gives a stronger brighter red stain. However we still see those crystals! We are suspecting a water problem. I have been dismantling some of the plumbing and getting some Ca crystals out of the pipes. We are manually coverslipping. ---Original Message--- From: Debra Sienadsi...@statlab.com To: 'e...@pigsqq.org'e...@pigsqq.org Subject: Re: [Histonet] annoying crystals on sections Sent: Aug 21 '12 07:46 Is your eosin alcoholic or aqueous? What is your staining protocol and what reagents are you using? This information would be most helpful. Also are the sections paraffin embedded and routinely processed in tissue processor? Debbie Siena HT(ASCP)QIHC Technical Manager | StatLab Medical Products 407 Interchange St. | McKinney, TX 75071 Direct: 972-436-1010 x229 | Fax: 972-436-1369 dsi...@statlab.com | www.statlab.com - Original Message - From: E. Wayne Johnson [mailto:e...@pigsqq.org] Sent: Monday, August 20, 2012 06:20 PM To: histonet@lists.utsouthwestern.eduhistonet@lists.utsouthwestern.edu Subject: [Histonet] annoying crystals on sections We are having problems with crystals precipitated on our slides which are HE stains on tissues from pigs. Tissues are fixed in buffered formalin. We had trouble months ago with formalin pigment and we had resolved that by using ammonia in EtOH or picric acid in EtOH. Sometimes we receive fixed samples from the field that are not buffered but presently all of our tissues are fixed in neutral phosphate buffered formalin. We moved the Sakura autostainer to a different location under a fume hood on a different floor of the building to get the solvent odor out of our work area. Immediately we began to see a tremendous degradation in slide quality due to what we initially thought was formalin pigment. We have changed all of the solutions and all of the stains. We find that if we
[Histonet] antigen retrieval
To those with the Biocare intelliPATH system, Is the antigen retrieval part of the IHC automated or do you need to use the Biocare decloaker? Thankyou, Tom Truscott ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Re: Histonet Digest, Vol 98, Issue 36
Hi Jennifer, I am by no means an expert on this, but have done a few mouse guts. I think that trying to flush out the feces with formalin or saline soon after necropsy, would help preserve the mucosa. The bacteria in the gut start breaking down the mucosa soon after death. Perhaps the gut is thin enough to fix rapidly enough to prevent damage to the mucosa without flushing. If that is the case, then flushing out the feces after fixation might help the quality of your slides. You may have to get permission to open up the gut to flush out the feces. It may hinge on how the tissue needs to be trimmed and oriented. Good luck, Tom Truscott -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jennifer Sipes Sent: Thursday, January 26, 2012 10:13 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Histonet Digest, Vol 98, Issue 36 Hi Histonetters! I have a question that I'm hoping you guys can help me with. I have a person who brings me mouse intestinal tract with fecal matter still in it. She can NOT remove the fecal matter before processing due to the fact that her lab is studying the mucus membrane. She had asked the lab that was doing this how they got such prestine sections while we are having a hard time with the sections staying neat due to all the nicks caused by the fecal matter. Does anyone have any suggestions as to how best to handle this?? It seems to be getting worse the further into the study she goes. Thank you in advance for any help that you give!! It is truly appreciated! Jennifer K. Sipes, ALAT Sr. Laboratory Technician Johns Hopkins University Ross 933 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 fax: 410-955-9677 cell: 443-631-6361 e-mail: jsip...@jhmi.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] RE: Interview Questions
Hi Sally, I think I would ask if they were familiar with the terms bovine, porcine, ovine, caprine, equine, ect. Then ask if they would process bovine tissue differently than mouse tissues. Ask what kind of problems they might expect to encounter doing IHC and immunofluorescence on animal tissue. If you handle legal cases, ask questions about tissue identity and chain of evidence. Ask questions on lab safety and chemical waste disposal. Good luck, Tom Truscott -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara Sent: Wednesday, January 25, 2012 7:37 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Interview Questions Okay, My People - I will be one of the interviewers for locating my replacement). I've not been this fortunate before and I do know there are questions one cannot ask so that's not an issue. What I'd like to know is what I SHOULD ask. This position is fairly straightforward - basic veterinary histology with nothing significantly challenging (but with that potential). What would YOU want to know about a candidate that would convince you that this person was The One? I need questions with meat to them. Your suggestions will be much-ly appreciated. Gracias! Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services 1101 Camino de Salud NE Albuquerque, NM 87102 505-383-9278 (Histology Lab) ___ Histonet mailing list Histonet@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: saffron vs. safran du gatinais
Hi Beth, Saffron is a great spice. We use it in a old family recipe from Cornwall for a bread roll( saffron nubbies). We also find the price very expensive, but also varies a lot. If you know anyone in the middle East, or India, they could get it a lot cheaper. We have raised it in our garden and get about four 3/4 in long stigmas from each flower, so it is labor intensive to get much, but a little goes a long way. It has to. I don't know of any substitute in staining. Tom Truscott -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Villarreal, Beth Sent: Thursday, January 05, 2012 12:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] saffron vs. safran du gatinais Hello histonet, I have a protocol that calls for safran du gatinais and am experiencing some serious sticker shock. Can I substitute saffron in my solution or am I asking for trouble? Many thanks, Beth Beth Villarreal Scientist I Novartis Institutes for BioMedical Research, Inc. 300 Technology Square Cambridge, MA 02139 USA ___ Histonet mailing list Histonet@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] Re: Histonet Digest, Vol 93, Issue 24
Or, has you tried gently infusing the intestinal lumen with formalin from a syringe and tying or clamping both ends, then allow to fix in a jar of formalin, then trim open lengthways, then trim C-shaped cross-sections, then remove fecal material before placing in cassette. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Grantham, Andrea L - (algranth) Sent: Thursday, August 18, 2011 12:13 PM Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: Histonet Digest, Vol 93, Issue 24 Has she tried opening the colons and allowing them to fix and then gently flushing the fecal matter out before processing? Andi On Aug 18, 2011, at 10:34 AM, Jennifer Sipes wrote: Hi everyone! First let me thank EVERYONE for the huge response on the HE stain on frozen tissue!!! My slides turned out great! Now, I've got a question that I'm thinking will stump you as much as it's stumped a co-worker and I! She is doing a study on the mucus membrane in the intestinal tract(cecum and colon mainly) of the mouse. She tried flushing the fecal matter out of the colon and cecum but lost the membrane. She decided to leave the fecal matter in, however now when I section it, it tears up my blade and gives sections that are not all that great. I was wondering if anyone knows how to acheive great sections while keeping the membrane? Thank you so much in advance for any and all help you can give us! Regards, Jen Jennifer K. Sipes, ALAT Sr. Laboratory Technician Johns Hopkins University Ross 933 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 fax: 410-955-9677 cell: 443-631-6361 e-mail: jsip...@jhmi.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] RE: Cresyl violet stain on 50 um mouse brain sections
Hi Susan, I would suggest cutting lots thinner sections. The plus slides seem to have trouble holding on to very thick sections. Another possibility is too much glacial acetic acid in your working solution of cresyl-too much acid can loosen the bonds on the plus slides. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Michael, Susan Sent: Friday, July 29, 2011 6:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cresyl violet stain on 50 um mouse brain sections I am having trouble keeping my sections on the slides when I stain with cresyl violet. These are fixed frozen sections, 50 um, dried overnight on plus slides, overnight in 1 to 1 alcohol/chloroform, then rehydrated through 100, 95 ETOHs then to water. Into the cresyl violet solution, then 100% to 95% ETOH. Is it the water? Is it the slides? Any suggestions? Susan ___ Histonet mailing list Histonet@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] comprehensive list?
Hi All, Is there a comprehensive list in one location somewhere that gives information on all or most antibodies on how they work with FFPE or FS or other fixatives, so that we don't have to try to many approaches when we get a new antibody to try. Thanks, Tom T ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: comprehensive list?
Yes, vendor sheets are good, but not all give enough info, and you may have to compare several vendor, and as you say, antibodies only tested by western blot are common in research. Yet, it would be a great money and timesaver if there was one site to go to, to see if anyone had tried these antibodies by different methods and could also submit their own results for entry, possibly with links to Journal articles, much like what some vendors do. It could be a quick reference guide to setting up some research or double staining methods, or just see if some new request is going to fit your routine. Someone would have to have more time and computer skills than I. Thanks all for the input, TOM T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Morken, Timothy Sent: Friday, March 18, 2011 10:38 AM To: Histonet Subject: [Histonet] RE: comprehensive list? Tom, Vendor datasheets are the best source of information. The vast majority of clinical vendors antibodies are designed for formalin fixed paraffin embedded tissue. Research vendors are all over the map and often only test by western blots or elisa so they may or may not have information about how their antibodies work in fixed tissue. Tim Morken Supervisor, Histology, IPOX UCSF Medical Center San Francisco, CA, USA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Truscott, Tom Sent: Friday, March 18, 2011 9:02 AM To: Histonet Subject: [Histonet] comprehensive list? Hi All, Is there a comprehensive list in one location somewhere that gives information on all or most antibodies on how they work with FFPE or FS or other fixatives, so that we don't have to try to many approaches when we get a new antibody to try. Thanks, Tom T ___ Histonet mailing list Histonet@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] Slide Labeling System that Survives Citrate Boiling
That sounds like the one TBS made or makes. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of louise renton Sent: Wednesday, January 05, 2011 10:39 PM To: Cameron Nowell; Histonet Subject: Re: [Histonet] Slide Labeling System that Survives Citrate Boiling long ago there was a cute system that worked with superfrost type slides - you know the ones that have the sort of coloured ends - etched through the paint to make a permanent marking Common HB pencil also survives on ordinary frosted slidesand surgipath pen marking esp if used on superfrost slides On 1/6/11, Cameron Nowell cameron.now...@ludwig.edu.au wrote: Hi List, I am looking for a slide labelling system to print labels for our research histology samples that will survive pretty much anything we throw at it. There seems to be lots of choices out there for chemical resistant labels but i can't seem to find much on ones that are resistant to antigen retrieval like citrate boiling. I have searched google and the histonet archives and the best i can find is a reference to General Data having some that may do the job. Does anyone out there have any more info or are you using someting that works well? Thanks Cam Cameron J. Nowell Microscopy Manager Centre for Advanced Microscopy Ludwig Institute for Cancer Research PO Box 2008 Royal Melbourne Hospital Victoria, 3050 AUSTRALIA Office: +61 3 9341 3155 Mobile: +61422882700 Fax: +61 3 9341 3104 Facility Website http://www.ludwig.edu.au/confocal/ This communication is intended only for the named recipient and may contain information that is confidential, legally privileged or subject to copyright; the Ludwig Institute for Cancer Research Ltd does not waive any rights if you have received this communication in error. The views expressed in this communication are those of the sender and do not necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd. ___ 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 +27 11 717 2298 (tel fax) 073 5574456 (emergencies only) 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. ___ Histonet mailing list Histonet@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] PLP fixative
I am seeking basic information on PLP fixative. What is it? Can I make it or buy it? How long does fixation take for lymph node? Thanks in advance, Tom Truscott ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: billing consults
One way to think about it, would be that if a pathologist accepts a case, it is understood that he or she has the expertise to diagnose it, and with a diagnoses comes a bill. But, if unable to diagnose, or confidently diagnose, and a consult is needed, then the pathologist that can make the diagnosis gets paid, and the lab that did the preliminary work gets part of that pay. Would a mechanic charge you full price for fixing your engine, if he couldn't fix it and sent it to another mechanic to fix? Perhaps fee schedules should be set up so that a certain percentage goes to consultation ( because it's going to happen) and the patient doesn't get extra billing.Just an opinion. Tom Truscott -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tench, Bill Sent: Wednesday, September 08, 2010 9:48 AM To: histo...@pathology.swmed.edu Subject: [Histonet] billing consults we ask the consultant to bill the patient's insurance. If they don't do that, they bill the hospital and the hospital passes the charges on to the patient. we do not make any distinction based on where the request for the consultation came from (us, the patient, the treating clinician). The patient is the beneficiary of the service. On the very rare case when it clearly is an issue of intellectual curiosity (i can think of only 3 examples), the practice will pay the charge. Bill Tench Associate Dir. Laboratory Services Chief, Cytology Services Palomar Medical Center 555 E. Valley Parkway Escondido, California 92025 bill.te...@pph.org Voice: 760- 739-3037 Fax: 760-739-2604 [None] made the following annotations - Confidential E-Mail: This e-mail is intended only for the person or entity to which it is addressed, and may contain information that is privileged, confidential, or otherwise protected from disclosure. Dissemination, distribution, or copying of this e-mail or the information herein by anyone other than the intended recipient is prohibited. If you have received this e-mail in error, please notify the sender by reply e-mail, and destroy the original message and all copies. - ___ Histonet mailing list Histonet@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] water bath information
Hi, The old TBS's are blue, and the new ones are white. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cynthia Pyse Sent: Tuesday, June 08, 2010 12:02 PM To: 'Cheryl'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] water bath information I am also looking for the same type of waterbaths and would appreciate any information. Thanks Cindy Pyse, CLT, HT (ASCP) Histology Supervisor X-Cell Laboratories e-mail cp...@x-celllab.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cheryl Sent: Tuesday, June 08, 2010 12:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] water bath information Hi Guys- We need new waterbaths and I'm looking for a specific type. It's blue with a square glass baking dish inset on the top. We don't care if they're blue but the small footprint with the glass insert is the key-- Vendors and refurb dealer responses welcome- Thanks! Cheryl Kerry, HT(ASCP) Houston ker...@labcorp.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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Contamination..processor?
If not already mentioned, wiping your heated forcep off with a kimwipe or paper towel before and after embedding each tissue helps prevent this problem at embedding. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of kim.dona...@bhcpns.org Sent: Wednesday, March 24, 2010 10:01 AM To: tigger...@aol.com Cc: histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Subject: Re: [Histonet] Contamination..processor? You are definite that the cells are in the block? If so I would change all the paraffins out as well, even the paraffin on your embedding center. Kim Donadio Pathology Supervisor Baptist Hospital 1000 W Moreno St. Pensacola FL 32501 Phone (850) 469-7718 Fax (850) 434-4996 tigger...@aol.com Sent by: histonet-boun...@lists.utsouthwestern.edu 03/24/2010 11:40 AM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Contamination..processor? Hello everyone. Today we have a problem with contamination. The pathologist notes cells from tonsil specimens here and there on our GI biopsy slides. The cells are in the block. I'm trying to ascertain the source of the contamination. The grossing pathologist grossed the tonsils AFTER all GI specimens yesterday (not source of contaminant). We (the techs) embedded all GI specimens first, trimmed, cut, floated and stained ALL GI specimens BEFORE the tonsils (not source of contaminant). The only other source of the contamination I can think of is from the tissue processor. We have a Tissue Tek VIP closed processor. Has anyone ever experienced any problems like this? We had a similar issue a few weeks ago. I thought the contaminant cells may be from a bladder tumor, which had multiple sections submitted. In this instance the cells showed up days work of the bladder tumor, and in the following days work also (though the pathologists could not say for sure the cells were from the bladder case). We changed our formalin solutions in the processor and the problem did not present the next day. We also started putting all bladder tumor specimens in the microcassettes, to prevent tissue from escaping. Has anyone had any problem like this, or does anyone have any ideas on how to prevent this in the future? We had not seen this problem until these past two incidences, and this tonsil problem is particularly strange to me because we process tonsils and GI specimens in the same workload on a regular basis and have never had this issue before. Any help is appreciated! Thanks! Brandi ___ 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Anything New for Finding Lymph Nodes in Colon Fat?
Hi Wanda, Soaking tissues in white vinegar tends to clear the fat and make the lymph nodes more visible, but it might affect some tests. Tom Truscott -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of dkb...@chs.net Sent: Monday, November 30, 2009 12:21 PM To: Smith Wanda Cc: histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Subject: Re: [Histonet] Anything New for Finding Lymph Nodes in Colon Fat? We use Lymph Node Revealing Solution from Polyscientific. Works great for us. Debbie M. Boyd, HT(ASCP) I Chief Histologist I Southside Regional Medical Center I 200 Medical Park Boulevard I Petersburg, Va. 23805 I T: 804-765-5050 I F: 804-765-5582 I dkb...@chs.net Smith Wanda wanda.sm...@hcahealthcare.com Sent by: histonet-boun...@lists.utsouthwestern.edu 11/30/2009 02:59 PM To histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu cc Subject [Histonet] Anything New for Finding Lymph Nodes in Colon Fat? Has anyone discovered anything new since the last discussion in the archives for finding lymph nodes in colonic fat??? Thanks, Wanda WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Disclaimer: This electronic message may contain information that is Proprietary, Confidential, or legally privileged or protected. It is intended only for the use of the individual(s) and entity named in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from your computer. Do not deliver, distribute or copy this message and do not disclose its contents or take any action in reliance on the information it contains. ___ Histonet mailing list Histonet@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] Import of staining reagents(richard allan/microm, now thermo)
Hi Abijag, You can get MSDS forms from the company that explains any hazards associated with these reagents or any others. You can study these with your local distributor. These reagents are packed by RA to be safely shipped anywhere in this country and I would assume wouldn't need much extra to go to India. Tom Truscott -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of abi jag Sent: Thursday, October 29, 2009 9:20 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Import of staining reagents(richard allan/microm, now thermo) Hello histonetters, This is with regard to import of Richard allan(microm, now thermo)staining reagents from US to India. After facing several problems from stains manufactured by local manufacturers, we thought of switching over to RA stains. The local distributor here is telling that these reagents(we are planning for procuring hematoxylin,eosin,bluing reagent and clarifier)are highly hazardous and need special packing(itself require 4000 US $). Anybody having any sort of experience about this and having idea about how RA stains can be transported/used in other parts of the world. Thanks for all your help abijag Try the new Yahoo! India Homepage. Click here. http://in.yahoo.com/trynew ___ Histonet mailing list Histonet@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] Floaters in Waterbath
Hi Stella, Not only wiping the top of the waterbath water with kimwipes between each block, and keeping forceps clean at embedding, and keeping your slides clean, but also keeping things clean at grossing: clean cutting board and instruments between tissues or cases. One pathologist called it forcep metastasis. Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stella Mireles Sent: Friday, October 23, 2009 7:11 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Floaters in Waterbath I know we have all had some problems with floaters in our waterbath at some point in our microtomy career. Our doctors are very picky and I need some tips on keeping an immaculate clean waterbath, but not sacrificing the speed in a regular routine lab. We use the pyrex waterbath and paper towels for wiping our area. 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
RE: [Histonet] PIN4
Hi Ann, Are you using the same microscope in the new location? Tom Truscott USDA-ARS -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of thisis...@aol.com Sent: Friday, February 06, 2009 1:09 PM To: histonet@lists.utsouthwestern.edu Cc: atiak...@qdxpath.com; ake...@qdxpath.com Subject: [Histonet] PIN4 We just moved a Ventana Benchmark XT to a new location.? It appears to be working exactly the same as it was at the old location...no issues.? We used the same IView DAB detection kit? (same? protocol as well), same control tissue and same Vulcan Fast Red reagents.? All procedures are followed exactly at both locations. Can someone tell me why the Vulcan Fast REd is staining with a brown color to it instead of red at the new location.? We even used distilled water for rinsing and decontaminated the XT.? I don't know what else to do Any ideas? Thanks, Ann ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
