RE: [Histonet] Best IHC stainer
I echo Joyce's comments! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 I find that the harder I work, the more luck I seem to have. - Thomas Jefferson Weems, Joyce jwe...@sjha.org 8/26/2009 10:57 PM They are both good instruments. From my experience pricing is high for the Ventana reagents and there seems to be an increase in DAKO pricing from what I see on the Net. Have you looked at the Leica Bond? We found that to be a good fit for us. Started out to add it to our DAKO line, but ended up changing completely. It also does ISH, and is a continuous feed - up to 10 slides at a time. Good luck, Joyce -Original Message- From: histonet-boun...@lists.utsouthwestern.edu on behalf of Gareth Blaeuer Davis Sent: Wed 8/26/2009 8:04 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Best IHC stainer The lab I work in has been doing demos on IHC stainers Ventana Benchmark XT and Dako's Autostainer Plus. We could really use feed back from current users of both. We are having a hard time deciding between the two, so any input would be great. What are the Pros and Cons with both. Thanks, Gareth Blaeuer Davis, B.S., HT _ Hotmail® is up to 70% faster. Now good news travels really fast. http://windowslive.com/online/hotmail?ocid=PID23391::T:WLMTAGL:ON:WL:en-US:WM_HYGN_faster:082009___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. Déclaration de confidentialité Le présent message (y compris les annexes) peut contenir des renseignements confidentiels à l'intention d'une personne ou d'un organisme particulier. Si vous avez reçu la présente communication par erreur, veuillez en informer l'expéditeur immédiatement. Si vous n'êtes pas le destinataire prévu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer immédiatement de votre système informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Double HRP immunostains
Andrea,Go for: Vector Blue (Vector Labs) or Permanent Blue (Diagnostic Biosystems) for AP activityVector Red or Permanent Red, also for AP activity. These two AP immunostaining methods can be combined in a sequential double staining method with a HIER step (10 min, 98C) in between for removing the immunoreagents. Both red or blue reaction products survive the HIER step without damage (see JOH 31:119-127). Cheers, ChrisChris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Wed, 26 Aug 2009 14:05:41 -0400 From: Andrea Hooper anh2...@med.cornell.edu Subject: [Histonet] Double HRP immunostains To: Histonet histonet@lists.utsouthwestern.edu I do double HRP immunostains. Works well using DAB/AEC and True Blue ... but what are your favorite chromagens to use? Would love some feedback for inspiration. Andrea -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Sakura Printers
Hi Joyce, Hi Pat, From my experience the printers are working very well. There could be issues if any of the adjustments are of or the slides or cassettes that are used not fit to the system. What kinds of problems do you having? What makes you unhappy with the printer? @Joyce: I would like to know what kind of LIS system do you using. Do you have the printers connected directly to a computer or do you using a network switch as an interface to the printers. Thanks Gregor ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Best IHC stainer
I prefer the DAKO. Check, besides the instrument itself, the reagents COST and how open one system is compared with the other. René J. --- On Wed, 8/26/09, Gareth Blaeuer Davis gareth.da...@hotmail.com wrote: From: Gareth Blaeuer Davis gareth.da...@hotmail.com Subject: [Histonet] Best IHC stainer To: histonet@lists.utsouthwestern.edu Date: Wednesday, August 26, 2009, 8:04 PM The lab I work in has been doing demos on IHC stainers Ventana Benchmark XT and Dako's Autostainer Plus. We could really use feed back from current users of both. We are having a hard time deciding between the two, so any input would be great. What are the Pros and Cons with both. Thanks, Gareth Blaeuer Davis, B.S., HT _ Hotmail® is up to 70% faster. Now good news travels really fast. http://windowslive.com/online/hotmail?ocid=PID23391::T:WLMTAGL:ON:WL:en-US:WM_HYGN_faster:082009___ 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] Confirmation of reports
We use Copath for Histology LIS and Epic as our Hospital and clinic based system. In Epic, you can retrieve who reviewed your results by looking at the result in Chart Review. Look below the results, it lists who and when. There is also an electronic trail that Epic can produce if needed. Dorothy Webb, HT Regions Histology Technical Supervisor 651-254-2962 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Organizing of cassettes for processing
We load the cassettes into the tissue processing basket as they are grossed in. They stay in a container of formalin until they are loaded on the tissue processor. What concern about formalin exposure do you have with this method? The gross hood is on while they are doing this. We also monitor our formalin exposure annually and I have the path assistant wear the formalin badge while she performs this task. Our readings have always been well below the acceptable range. If there is a better method I may be interested in it also. Thanks, Carol -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of ba...@gundluth.org Sent: Wednesday, August 26, 2009 4:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Organizing of cassettes for processing Hi all - For those that do batch overnight processing, how do you organize the cassettes? Currently we have 2 path assistants that gross throughout the day, and each puts their cassettes as they are grossed into a bucket of formalin. At the end of the day a histotech drains the formalin off, rinses the cassettes in water, then manually puts the cassettes into order according to our worklist, with rush cases being put up front. The baskets are then loaded onto the tissue processor (Sakura VIP5 and VIP6). We are wondering if there are some other ideas of how to streamline this process. One thought was to have the cassettes loaded/organized into the tissue processing basket as they are grossed, but have a concern about formalin exposure while doing this. Any thoughts would be greatly appreciated. Barb Moe Gundersen Lutheran Medical Center La Crosse WI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet NOTICE OF CONFIDENTIALITY This message, including any attachments, is intended only for the sole use of the addressee and may contain confidential or privileged information that is protected by the State of Kentucky and/or Federal regulations. If you are not the intended recipient, do not read, copy, retain or disseminate this message or any attachment. If you have received this message in error, please call the sender immediately at (859)258-4000 and delete all copies of this message and any attachment. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. Neither the transmission of this message or any attachment, nor any error in transmission or misdelivery shall constitute waiver of any applicable legal privilege. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Best IHC stainer
Ventana reagents eem to be a little pricey, but if you want a system that you can load and walk away from, it is the way to go. DAKO is an easy enough to use instrument and is an open system, but they are going to a price per slide kind of thing, if I remmeber correctly. I prefer the IP from Biocare, as is is an open system and a continuous load instrument. And it mixes the chromagen online -Original Message- From: Gareth Blaeuer Davis gareth.da...@hotmail.com To: histonet@lists.utsouthwestern.edu Sent: Wed, Aug 26, 2009 8:04 pm Subject: [Histonet] Best IHC stainer he lab I work in has been doing demos on IHC stainers Ventana Benchmark XT and ako's Autostainer Plus. We could really use feed back from current users of both. We are having a hard time deciding between the two, o any input would be great. What are the Pros and Cons with both. Thanks, Gareth Blaeuer Davis, B.S., HT _ otmail® is up to 70% faster. Now good news travels really fast. ttp://windowslive.com/online/hotmail?ocid=PID23391::T:WLMTAGL:ON:WL:en-US:WM_HYGN_faster:082009___ istonet mailing list isto...@lists.utsouthwestern.edu ttp://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Best IHC stainer
Agreed. Intellipath from Biocare. Load it and walk away - and it has a voice notification to tell you the run is finished. You can load it in the PM, and take slides off in the AM. Love it. Has a chilling station for AP chromogen, too. Jacke O' godsgal...@aol.com Sent by: histonet-boun...@lists.utsouthwestern.edu 08/27/2009 10:05 AM To gareth.da...@hotmail.com, histonet@lists.utsouthwestern.edu cc Subject Re: [Histonet] Best IHC stainer Ventana reagents eem to be a little pricey, but if you want a system that you can load and walk away from, it is the way to go. DAKO is an easy enough to use instrument and is an open system, but they are going to a price per slide kind of thing, if I remmeber correctly. I prefer the IP from Biocare, as is is an open system and a continuous load instrument. And it mixes the chromagen online -Original Message- From: Gareth Blaeuer Davis gareth.da...@hotmail.com To: histonet@lists.utsouthwestern.edu Sent: Wed, Aug 26, 2009 8:04 pm Subject: [Histonet] Best IHC stainer he lab I work in has been doing demos on IHC stainers Ventana Benchmark XT and ako's Autostainer Plus. We could really use feed back from current users of both. We are having a hard time deciding between the two, o any input would be great. What are the Pros and Cons with both. Thanks, Gareth Blaeuer Davis, B.S., HT _ otmail® is up to 70% faster. Now good news travels really fast. ttp://windowslive.com/online/hotmail?ocid=PID23391::T:WLMTAGL:ON:WL:en-US:WM_HYGN_faster:082009___ istonet mailing list isto...@lists.utsouthwestern.edu ttp://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] Best IHC stainer
Having worked with Dako autostainer for years and currently using Vantana Discovery XT and Leica Bond stainer's, my preference would be the Bond. You have the capability of the Ventana stainer, and being able to unlock the software that will allow you to use it as any other stainer on the market today. Bond also has a favorable per slide cost analysis. If you need any additional info, I would be happy to provide as much as I am can. Thanks Dusko Trajkovic Pfizer Inc La Jolla 858-638-6202 From: godsgal...@aol.com godsgal...@aol.com To: gareth.da...@hotmail.com; histonet@lists.utsouthwestern.edu Sent: Thursday, August 27, 2009 8:05:35 AM Subject: Re: [Histonet] Best IHC stainer Ventana reagents eem to be a little pricey, but if you want a system that you can load and walk away from, it is the way to go. DAKO is an easy enough to use instrument and is an open system, but they are going to a price per slide kind of thing, if I remmeber correctly. I prefer the IP from Biocare, as is is an open system and a continuous load instrument. And it mixes the chromagen online -Original Message- From: Gareth Blaeuer Davis gareth.da...@hotmail.com To: histonet@lists.utsouthwestern.edu Sent: Wed, Aug 26, 2009 8:04 pm Subject: [Histonet] Best IHC stainer he lab I work in has been doing demos on IHC stainers Ventana Benchmark XT and ako's Autostainer Plus. We could really use feed back from current users of both. We are having a hard time deciding between the two, o any input would be great. What are the Pros and Cons with both. Thanks, Gareth Blaeuer Davis, B.S., HT _ otmail® is up to 70% faster. Now good news travels really fast. ttp://windowslive.com/online/hotmail?ocid=PID23391::T:WLMTAGL:ON:WL:en-US:WM_HYGN_faster:082009___ istonet mailing list isto...@lists.utsouthwestern.edu ttp://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
AW: [Histonet] Organizing of cassettes for processing
We seperate biopsies and grossed tissue before grossing. They get into differently coloured cassettes, and different VIPs. The cassettes of the grossed tissue are put directly into the VIP baskets - short time dry, then put into a container with formalin. While grossing a report is written. So the order of the cassettes is mostly the same as the numbers on the report. Urgent cases have an extra colour. After embedding, the cassettes and the report are checked. Urgent cases are embedded first, then biopsies, grossed tissue is sorted according to the report not numbers. With slide-delivering we check sildes and report again. So mislabelled or missing cassettes and slides are identified. Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von ba...@gundluth.org Gesendet: Mittwoch, 26. August 2009 22:38 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Organizing of cassettes for processing Hi all - For those that do batch overnight processing, how do you organize the cassettes? Currently we have 2 path assistants that gross throughout the day, and each puts their cassettes as they are grossed into a bucket of formalin. At the end of the day a histotech drains the formalin off, rinses the cassettes in water, then manually puts the cassettes into order according to our worklist, with rush cases being put up front. The baskets are then loaded onto the tissue processor (Sakura VIP5 and VIP6). We are wondering if there are some other ideas of how to streamline this process. One thought was to have the cassettes loaded/organized into the tissue processing basket as they are grossed, but have a concern about formalin exposure while doing this. Any thoughts would be greatly appreciated. Barb Moe Gundersen Lutheran Medical Center La Crosse WI ___ 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
AW: [Histonet] DIF tissue in GLUT
Anne, we do DIF on formalin fixed renal biopsies routinly. To get good results we perform Protease digestion before DIF. I don't have the protocol here, but we use Proteinase K from Dako at RT for 3-4 min, and the antibodies are between 1:20 and 1:40 für 30 min. I don't know, if that would help with glutaraldehyde-fixed specimen, but worth trying. Gudrun Lang Akh Linz, Austria -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Anne van Binsbergen Gesendet: Donnerstag, 27. August 2009 12:24 An: John Kiernan; histonet@lists.utsouthwestern.edu Betreff: [Histonet] DIF tissue in GLUT Dear John and other knowledgeable Histonetters and Gurus of the trade Renal bx tissue core has been placed in Gluteraldehyde/Trumps EM fixative by mistake (we all make mistakes) - we now need to do DIF Does anyone out there have a method for this? -- Anne van Binsbergen (Hope) Abu Dhabi UAE ___ 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] Michael LaFriniere
If anyone knows how to contact Michael, please email me. Thanks! Phyllis Thaxton HT(ASCP)QIHC DCH Regional Medical Center Tuscaloosa, AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [SPAM-HC] - [Histonet] Organizing of cassettes for processing - Email found in subject
I gave out misinformation which I want to rectify. The metal sleeves and holding tray is sold by McCormick Scientific which is a part of Leica, but they have their own ordering website: http://www.mccormickscientific.com/cassettemangement.asp?PCID=853 The above link should open to the item itself. The ordering information is- # MC 100 Formalin Holding tank # MC 101 Formalin Tank Lid Ruth Cazares, HT (ASCP) Histology Supervisor Department of Pathology Swedish Covenant Hospital 5145 North California Ave Chicago, IL 60625 -Original Message- From: Cazares, Ruth Sent: Wednesday, August 26, 2009 4:22 PM To: 'ba...@gundluth.org'; histonet@lists.utsouthwestern.edu Subject: RE: [SPAM-HC] - [Histonet] Organizing of cassettes for processing - Email found in subject Hi Barb, We have metal sleeves in which 18 cassettes fit standing front to back, and a metal box with lid which we fill with formalin. This box holds 6 sleeves and as cases are grossed, the cassettes are picked up in order (it doesn't take any effort to do this at the time of grossing) and placed in the sleeves. We have two metal box/containers in which we separate our routines and our biopsies (we use a short program for our biopsies). At the end of the day we print out hard copies of all the cases that will be cut the next morning, separating the routines from the biopsies so there are 2 lists, and then we go cassette by cassete and check off on the list to assure that every cassette is accounted for. This system works great for us and it helps to find errors before they go any further. We use colored dots on the cases that are biopsies so when its time to print out our log sheets we can do so from the requisitions or working log sheet, and if done right, every case and cassette should match up. I believe Leica now carries these sleeves and metal box containers, I strongly urge you to look into them, they keep things so organized and we LOVE them!! I can get you ordering information, just let me know. Ruth Cazares, HT (ASCP) Histology Supervisor Department of Pathology Swedish Covenant Hospital 5145 North California Ave Chicago, IL 60625 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of ba...@gundluth.org Sent: Wednesday, August 26, 2009 3:38 PM To: histonet@lists.utsouthwestern.edu Subject: [SPAM-HC] - [Histonet] Organizing of cassettes for processing - Email found in subject Hi all - For those that do batch overnight processing, how do you organize the cassettes? Currently we have 2 path assistants that gross throughout the day, and each puts their cassettes as they are grossed into a bucket of formalin. At the end of the day a histotech drains the formalin off, rinses the cassettes in water, then manually puts the cassettes into order according to our worklist, with rush cases being put up front. The baskets are then loaded onto the tissue processor (Sakura VIP5 and VIP6). We are wondering if there are some other ideas of how to streamline this process. One thought was to have the cassettes loaded/organized into the tissue processing basket as they are grossed, but have a concern about formalin exposure while doing this. Any thoughts would be greatly appreciated. Barb Moe Gundersen Lutheran Medical Center La Crosse WI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet histonet *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Organizing of cassettes for processing
We do the same and all of our readings have been well under what they should be. Dawn D. Schneider, HT(ASCP) Howard Young Medical Center Woodruff, WI 54568 (715)356-8174 dawn.schnei...@ministryhealth.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Carol Bryant Sent: Thursday, August 27, 2009 8:56 AM To: 'ba...@gundluth.org'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Organizing of cassettes for processing We load the cassettes into the tissue processing basket as they are grossed in. They stay in a container of formalin until they are loaded on the tissue processor. What concern about formalin exposure do you have with this method? The gross hood is on while they are doing this. We also monitor our formalin exposure annually and I have the path assistant wear the formalin badge while she performs this task. Our readings have always been well below the acceptable range. If there is a better method I may be interested in it also. Thanks, Carol -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of ba...@gundluth.org Sent: Wednesday, August 26, 2009 4:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Organizing of cassettes for processing Hi all - For those that do batch overnight processing, how do you organize the cassettes? Currently we have 2 path assistants that gross throughout the day, and each puts their cassettes as they are grossed into a bucket of formalin. At the end of the day a histotech drains the formalin off, rinses the cassettes in water, then manually puts the cassettes into order according to our worklist, with rush cases being put up front. The baskets are then loaded onto the tissue processor (Sakura VIP5 and VIP6). We are wondering if there are some other ideas of how to streamline this process. One thought was to have the cassettes loaded/organized into the tissue processing basket as they are grossed, but have a concern about formalin exposure while doing this. Any thoughts would be greatly appreciated. Barb Moe Gundersen Lutheran Medical Center La Crosse WI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet NOTICE OF CONFIDENTIALITY This message, including any attachments, is intended only for the sole use of the addressee and may contain confidential or privileged information that is protected by the State of Kentucky and/or Federal regulations. If you are not the intended recipient, do not read, copy, retain or disseminate this message or any attachment. If you have received this message in error, please call the sender immediately at (859)258-4000 and delete all copies of this message and any attachment. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. Neither the transmission of this message or any attachment, nor any error in transmission or misdelivery shall constitute waiver of any applicable legal privilege. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipient(s) named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please notify the sender at the electronic mail address noted above and destroy all copies of this communication and any attachments. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: DIF tissue in GLUT
Anne, Tissue that has been fixed in glutaraldehyde has very, VERY bright autofluorescence. Unless there is some way to minimize this (none that I'm aware of), your immunofluorescence will be impossible to read over the background signal. Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Best IHC stainer
I was a DAKO fan for years... But after using Ventana, I completely changed my mind! Sure you pay a little more, but honestly the amount is negligible! In fact, after pricing out Dako versus Ventana, in the end the was really no difference. Plus the quality of the Ventana is exceptional. No one can touch their antibodies... If they don't have the antibody, Cell Marque does. (Cell Marque even puts the antibody into a Ventana style dispenser so that you register it and run!!!) Who else does that? No one. Trust me, I have checked. OMG... now I am sounding like and ad... didn't mean too! I think Ventana is the best and you can't beat their customer service dept.. Dako falls short on tech hotline help Ventana will even try to help you, even if you have a Dako! I am just saying. Maria Katleba HT(ASCP) Ms Napa CA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of godsgal...@aol.com Sent: 27 August 2009 08:06 To: gareth.da...@hotmail.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Best IHC stainer Ventana reagents eem to be a little pricey, but if you want a system that you can load and walk away from, it is the way to go. DAKO is an easy enough to use instrument and is an open system, but they are going to a price per slide kind of thing, if I remmeber correctly. I prefer the IP from Biocare, as is is an open system and a continuous load instrument. And it mixes the chromagen online -Original Message- From: Gareth Blaeuer Davis gareth.da...@hotmail.com To: histonet@lists.utsouthwestern.edu Sent: Wed, Aug 26, 2009 8:04 pm Subject: [Histonet] Best IHC stainer he lab I work in has been doing demos on IHC stainers Ventana Benchmark XT and ako's Autostainer Plus. We could really use feed back from current users of both. We are having a hard time deciding between the two, o any input would be great. What are the Pros and Cons with both. Thanks, Gareth Blaeuer Davis, B.S., HT _ otmail® is up to 70% faster. Now good news travels really fast. ttp://windowslive.com/online/hotmail?ocid=PID23391::T:WLMTAGL:ON:WL:en-US:WM_HYGN_faster:082009___ istonet mailing list isto...@lists.utsouthwestern.edu ttp://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Notice from St. Joseph Health System: Please note that the information contained in this message may be privileged and confidential and protected from disclosure. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Re: DIF tissue in GLUT
I suggest you use a counterstain for your IF to reduce the autofluoresence. Evans Blue - the product can be used as a counterstain in immunohistochemistry when using FITC. After staining for immunofluorescence, dip sections in a 0.1% (w/v) in water solution of Evans Blue for 5-10 minutes. Rinse well in fresh PBS or water before coverslipping. Reference: Immunocytochemistry, Theory and Practice, p. 82 (1988). Purchase from Sigma-Aldrich. William DeSalvo, B.S., HTL(ASCP) From: t...@stowers.org To: histonet@lists.utsouthwestern.edu Date: Thu, 27 Aug 2009 12:18:47 -0500 Subject: [Histonet] Re: DIF tissue in GLUT Anne, Tissue that has been fixed in glutaraldehyde has very, VERY bright autofluorescence. Unless there is some way to minimize this (none that I'm aware of), your immunofluorescence will be impossible to read over the background signal. Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _ Windows Live: Keep your friends up to date with what you do online. http://windowslive.com/Campaign/SocialNetworking?ocid=PID23285::T:WLMTAGL:ON:WL:en-US:SI_SB_online:082009___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Toluidine Blue
I am currently writing a procedure for the use of Toluidine Blue staining for both FNA's and frozen sections at the request of one of our pathologists. Can anyone guide me in the use of this stain in lieu of Diff Quick or HE for rapid processing (such as references I could use, etc) and does anyone have a procedure I can cite for permanent mounting/storage of these slides? Thanks in advance, Edie Lehman MT(ASCP) Anatomical Pathology Supervisor ed...@fmchealth.org Fairfield Medical Center People you know. Care you trust. Visit us at http://www.fmchealth.org or our online store at http://fairfield.thehospitalstore.com Confidentiality Notice: This e-mail message, including any attachments is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review; use; disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] DIF tissue in GLUT
Dear Anne, I assume the glutaraldehyde tissue has been processed and embedded in paraffin. Following removal of paraffin and any pretreatment to unmask the antigen, you can block autofluorescence or change the color by treating the sections with 0.05% sodium borohydride prepared in PBS for 30 minutes. After the borohydride treatment, carefully wash the slides with multiple changes of PBS for thirty minutes. I would also use a good quality silane treated slide for maximum tissue adhesion. A comment of caution: sodium borohydride is poisonous. Be extremely careful with the powder. Reference for NaBH4: Embedment in Glycol Methacrylate at Low Temperature Allows Immunofluorescent Localization of a Labile Tissue Protein. JA Carnegie, ME McCully and HA Robertson, Journal of Histochemistry and Cytochemistry, Vol. 28, No. 4, pp 308-310, 1980 Methods in Cell Biology, by Leslie Wilson, page 117, section VI, Use of Glutaraldehyde in Immunofluorescence Microscopy. Academic Press 1982 I used this method in a double label fluorescence project a few years ago with excellent success. 23. Endothelial Metaplasia in the Iridocorneal Endothelial (ICE) Syndrome. DN Howell, T Damms, JL Burchette, JR Ross, WR Green. Investigative Ophthalmology and Visual Science, Vol. 38, No. 9, pages 1896-1901, August 1997. Best Regards, Jim Jim Burchette, HT(ASCP) QIHC A simple histotech from a little country hospital in North Carolina Anne van Binsbergen anni...@gmail.com Sent by: histonet-boun...@lists.utsouthwestern.edu 08/27/2009 06:27 AM To John Kiernan jkier...@uwo.ca, histonet@lists.utsouthwestern.edu Histonet@lists.utsouthwestern.edu cc Subject [Histonet] DIF tissue in GLUT Dear John and other knowledgeable Histonetters and Gurus of the trade Renal bx tissue core has been placed in Gluteraldehyde/Trumps EM fixative by mistake (we all make mistakes) - we now need to do DIF Does anyone out there have a method for this? -- Anne van Binsbergen (Hope) Abu Dhabi UAE ___ 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] (no subject)
I was wondering if anyone on the histonet uses an alcian yellow stain method for H. Pylori? Do you have a problem finding the alcian yellow, or do you use a substitute? I would be interested to hear your pros and cons on this stain. Do you stain manually or use a kit? If a kit, where do you purchase yours from? Thank you all for your insight and knowledge concerning this task. Deanne Knutson Anatomic Pathology Supervisor St. Alexius Medical Center 900 E. Broadway Bismarck, North Dakota 58506 (701)-530-6730 dknut...@primecare.org mailto:dknut...@primecare.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] (no subject)
We purchase our Alcian Yellow kit for Helicobacter from Newcomer Supply, Kit #9130. Phone number is 800-383-7799. They are a wonderful company to deal with and their Alcian Yellow kit works beautifully. Our pathologists love it!! Lynne Bell, HT (ASCP) Lead Histologist Central Vermont Medical Center 130 Fisher Road Barre, VT 05641 802-371-4923 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Re: DIF tissue in GLUT
William, thanks for the reminder, I had totally forgotten about using a counterstain to counteract AF in aldehyde fixed samples. We have some glutaraldehyde fixed cryosections of kidney I think I'll try this with. I can assure you, the autofluorescence in these samples is magnificent! I worry about using sodium borohydride because they are cryosections and I've heard the treatment can be a bit harsh on tissue. Here's some additional information since we're on the subject: http://www.uhnresearch.ca/facilities/wcif/PDF/Autofluorescence.pdf Kind regards, Teri -Original Message- From: WILLIAM DESALVO [mailto:wdesalvo@hotmail.com] Sent: Thursday, August 27, 2009 12:51 PM To: Johnson, Teri; histonet Subject: RE: [Histonet] Re: DIF tissue in GLUT I suggest you use a counterstain for your IF to reduce the autofluoresence. Evans Blue - the product can be used as a counterstain in immunohistochemistry when using FITC. After staining for immunofluorescence, dip sections in a 0.1% (w/v) in water solution of Evans Blue for 5-10 minutes. Rinse well in fresh PBS or water before coverslipping. Reference: Immunocytochemistry, Theory and Practice, p. 82 (1988). Purchase from Sigma-Aldrich. William DeSalvo, B.S., HTL(ASCP) From: t...@stowers.org To: histonet@lists.utsouthwestern.edu Date: Thu, 27 Aug 2009 12:18:47 -0500 Subject: [Histonet] Re: DIF tissue in GLUT Anne, Tissue that has been fixed in glutaraldehyde has very, VERY bright autofluorescence. Unless there is some way to minimize this (none that I'm aware of), your immunofluorescence will be impossible to read over the background signal. Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Windows Live: Keep your friends up to date with what you do online. Find out more.http://windowslive.com/Campaign/SocialNetworking?ocid=PID23285::T:WLMTAGL:ON:WL:en-US:SI_SB_online:082009 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] uneven alternating sections on cryostat
When cutting PFA fixed, cryoprotected tissue on our cryostat, I frequently find that every other section gets cut improperly. I'll get one nicely cut section and then on the next pass I only get half of a section. This cycle simply repeats over and over and I lose many slices. It has been a while since our cryostat has been serviced, so I'm wondering if this an operator error or a machine problem. I thought maybe the tissue temperature hadn't settled properly, but the uneven cutting still happens even when I let the tissue sit for 30 minutes in the chuck holder. (cutting mouse spinal cord at -20C) On another note, if anyone has any tips for improving white/gray matter contrast in frozen spinal cord sections stained with luxol fast blue, I'd be very appreciative. I do defat the slices with chloroform and differentiate with lithium carbonate but everything is usually either very dark or very light. I am learning as I go (checking the archives here often) so any help would be great. Thanks! Nathan Cramer Neurobiology and Behavior Cornell University Ithaca, NY 14853 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] uneven alternating sections on cryostat
Nathan, It sounds like the blade holder is not secure enough or even in the tightness on each side. Hope this helps. Robyn Date: Thu, 27 Aug 2009 16:44:11 -0400 From: natec...@gmail.com To: histonet@lists.utsouthwestern.edu Subject: [Histonet] uneven alternating sections on cryostat When cutting PFA fixed, cryoprotected tissue on our cryostat, I frequently find that every other section gets cut improperly. I'll get one nicely cut section and then on the next pass I only get half of a section. This cycle simply repeats over and over and I lose many slices. It has been a while since our cryostat has been serviced, so I'm wondering if this an operator error or a machine problem. I thought maybe the tissue temperature hadn't settled properly, but the uneven cutting still happens even when I let the tissue sit for 30 minutes in the chuck holder. (cutting mouse spinal cord at -20C) On another note, if anyone has any tips for improving white/gray matter contrast in frozen spinal cord sections stained with luxol fast blue, I'd be very appreciative. I do defat the slices with chloroform and differentiate with lithium carbonate but everything is usually either very dark or very light. I am learning as I go (checking the archives here often) so any help would be great. Thanks! Nathan Cramer Neurobiology and Behavior Cornell University Ithaca, NY 14853 ___ 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] Special stains and microwave
Greetings Histonetters, We have finally come to the point where we need to fish or cut bait in relation to purchasing a laboratory approved microwave and venting it. While I think it would be cool to have one, I'm wondering if the cost is justified to do a handful of special stains that could be done (though more slowly) in a laboratory oven like the one we already have. Anyone with the time or desire to opine, please chime in. Those who have purchased such a microwave, I'll gladly take suggestions. Thanks in advance William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Best IHC stainer Ventana Ab's
Everyone may want to check this out. Heard today that this is no longer the case. Maybe Ventana will make an announcement! -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Akemi Allison-Tacha Sent: Thursday, August 27, 2009 16:46 To: Maria Katleba Cc: histonet Subject: RE: [Histonet] Best IHC stainer Ventana Ab's Most people are unaware of this, but that's probably because Ventana has had a contract with Cell Marque for years as their primary antibody and other consumables source. --- On Thu, 8/27/09, Maria Katleba maria.katl...@stjoe.org wrote: From: Maria Katleba maria.katl...@stjoe.org Subject: RE: [Histonet] Best IHC stainer To: godsgal...@aol.com godsgal...@aol.com, gareth.da...@hotmail.com gareth.da...@hotmail.com, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Date: Thursday, August 27, 2009, 10:40 AM I was a DAKO fan for years... But after using Ventana, I completely changed my mind! Sure you pay a little more, but honestly the amount is negligible! In fact, after pricing out Dako versus Ventana, in the end the was really no difference. Plus the quality of the Ventana is exceptional. No one can touch their antibodies... If they don't have the antibody, Cell Marque does. (Cell Marque even puts the antibody into a Ventana style dispenser so that you register it and run!!!) Who else does that? No one. Trust me, I have checked. OMG... now I am sounding like and ad... didn't mean too! I think Ventana is the best and you can't beat their customer service dept.. Dako falls short on tech hotline help Ventana will even try to help you, even if you have a Dako! I am just saying. Maria Katleba HT(ASCP) Ms Napa CA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of godsgal...@aol.com Sent: 27 August 2009 08:06 To: gareth.da...@hotmail.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Best IHC stainer Ventana reagents eem to be a little pricey, but if you want a system that you can load and walk away from, it is the way to go. DAKO is an easy enough to use instrument and is an open system, but they are going to a price per slide kind of thing, if I remmeber correctly. I prefer the IP from Biocare, as is is an open system and a continuous load instrument. And it mixes the chromagen online -Original Message- From: Gareth Blaeuer Davis gareth.da...@hotmail.com To: histonet@lists.utsouthwestern.edu Sent: Wed, Aug 26, 2009 8:04 pm Subject: [Histonet] Best IHC stainer he lab I work in has been doing demos on IHC stainers Ventana Benchmark XT and ako's Autostainer Plus. We could really use feed back from current users of both. We are having a hard time deciding between the two, o any input would be great. What are the Pros and Cons with both. Thanks, Gareth Blaeuer Davis, B.S., HT _ otmail® is up to 70% faster. Now good news travels really fast. ttp://windowslive.com/online/hotmail?ocid=PID23391::T:WLMTAGL:ON:WL:en-US:WM_HYGN_faster:082009___ istonet mailing list isto...@lists.utsouthwestern.edu ttp://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Notice from St. Joseph Health System: Please note that the information contained in this message may be privileged and confidential and protected from disclosure. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Special stains and microwave
Bill, I can highly recommend the EBS processor microwave. At a base price of 8,900 (last I checked) it is one of the lowest cost processors on the market. I don't know if having the flexibility to process in this new microwave may help justify the price? I purchased mine from Stat Lab, and the customer service from the folks at EBS was outstanding; they even had a prob and air agitator block style holder designed and built for me when I suggested that it should come with one. I really like our model, and it offers great flexibility in our lab. Rosa Fields, HT (ASCP) Gastroenterology Specialties Histology Supervisor 4545 R Street Lincoln, NE 68503 402-465-4545 rfie...@gidocs.net The information contained in the message and the documents accompanying this message contain information that is privileged and confidential and is intended only for the use of the individual or entity named above. If the reader of this message is 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 communication, other than its return to the sender, is strictly prohibited. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of O'Donnell, Bill Sent: Thursday, August 27, 2009 4:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Special stains and microwave Greetings Histonetters, We have finally come to the point where we need to fish or cut bait in relation to purchasing a laboratory approved microwave and venting it. While I think it would be cool to have one, I'm wondering if the cost is justified to do a handful of special stains that could be done (though more slowly) in a laboratory oven like the one we already have. Anyone with the time or desire to opine, please chime in. Those who have purchased such a microwave, I'll gladly take suggestions. Thanks in advance William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] uneven alternating sections on cryostat
Thanks, Robyn... I'll make sure the holder is clamped down. (sometimes its the little things...) Best Nate R J VAZQUEZ wrote: Nathan, It sounds like the blade holder is not secure enough or even in the tightness on each side. Hope this helps. Robyn Date: Thu, 27 Aug 2009 16:44:11 -0400 From: [1]natec...@gmail.com To: [2]histo...@lists.utsouthwestern.edu Subject: [Histonet] uneven alternating sections on cryostat When cutting PFA fixed, cryoprotected tissue on our cryostat, I frequently find that every other section gets cut improperly. I'll get one nicely cut section and then on the next pass I only get half of a section. This cycle simply repeats over and over and I lose many slices. It has been a while since our cryostat has been serviced, so I'm wondering if this an operator error or a machine problem. I thought maybe the tissue temperature hadn't settled properly, but the uneven cutting still happens even when I let the tissue sit for 30 minutes in the chuck holder. (cutting mouse spinal cord at -20C) On another note, if anyone has any tips for improving white/gray matter contrast in frozen spinal cord sections stained with luxol fast blue, I'd be very appreciative. I do defat the slices with chloroform and differentiate with lithium carbonate but everything is usually either very dark or very light. I am learning as I go (checking the archives here often) so any help would be great. Thanks! Nathan Cramer Neurobiology and Behavior Cornell University Ithaca, NY 14853 ___ Histonet mailing list [3]histo...@lists.utsouthwestern.edu [4]http://lists.utsouthwestern.edu/mailman/listinfo/histonet References 1. mailto:natec...@gmail.com 2. mailto:histonet@lists.utsouthwestern.edu 3. mailto:Histonet@lists.utsouthwestern.edu 4. http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] uneven alternating sections on cryostat
I was having this problem too, but I think I finally figured it out. There is a screw on our cryostat that attaches the chuck to the rest of the machine. This screws fits into a small hole in the chuck. Sometimes the screw isn't well set in the hole or is well set but for some reason comes loose and causes the chuck to wobble. For some reason, this causes your problem. Now I make sure that everything is tightly secured, and I rarely have this problem. Good luck, Adam On Thu, Aug 27, 2009 at 4:47 PM, Nathan Cramer natec...@gmail.com wrote: Thanks, Robyn... I'll make sure the holder is clamped down. (sometimes its the little things...) Best Nate R J VAZQUEZ wrote: Nathan, It sounds like the blade holder is not secure enough or even in the tightness on each side. Hope this helps. Robyn Date: Thu, 27 Aug 2009 16:44:11 -0400 From: [1]natec...@gmail.com To: [2]histo...@lists.utsouthwestern.edu Subject: [Histonet] uneven alternating sections on cryostat When cutting PFA fixed, cryoprotected tissue on our cryostat, I frequently find that every other section gets cut improperly. I'll get one nicely cut section and then on the next pass I only get half of a section. This cycle simply repeats over and over and I lose many slices. It has been a while since our cryostat has been serviced, so I'm wondering if this an operator error or a machine problem. I thought maybe the tissue temperature hadn't settled properly, but the uneven cutting still happens even when I let the tissue sit for 30 minutes in the chuck holder. (cutting mouse spinal cord at -20C) On another note, if anyone has any tips for improving white/gray matter contrast in frozen spinal cord sections stained with luxol fast blue, I'd be very appreciative. I do defat the slices with chloroform and differentiate with lithium carbonate but everything is usually either very dark or very light. I am learning as I go (checking the archives here often) so any help would be great. Thanks! Nathan Cramer Neurobiology and Behavior Cornell University Ithaca, NY 14853 ___ Histonet mailing list [3]histo...@lists.utsouthwestern.edu [4]http://lists.utsouthwestern.edu/mailman/listinfo/histonet References 1. mailto:natec...@gmail.com 2. mailto:histonet@lists.utsouthwestern.edu 3. mailto:Histonet@lists.utsouthwestern.edu 4. 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] uneven alternating sections on cryostat
Nathan: As indicated by others - first check that everything is tight as that is most often the issue - don't forget to check the adhesion of the specimen to the chuck as well. Also check the angle of approach to the blade as it could be too steep or too shallow. If everything is tight and properly adjusted, then you may have a retraction issue. If your cryostat retracts during the upstroke, the head may still be returning to the proper distance from the head to the specimen during the downstroke and hence you may see no section at first and then a gradually increasing thickness of section during the remainder of the downstroke. This can be due to two causes: 1) You can make this happen by operating the microtome at too high of a speed, if you slow down and the process improves then speed may be a factor; 2) Your microtome may be sticky and require maintenance (defrosting, cleaning, lubrication, etc.). The latter is best left to someone with extensive experience or a service rep. Good luck. Joe Galbraith University of Iowa joseph-galbra...@uiowa.edu -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adam . Sent: Thursday, August 27, 2009 4:53 PM To: Nathan Cramer Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] uneven alternating sections on cryostat I was having this problem too, but I think I finally figured it out. There is a screw on our cryostat that attaches the chuck to the rest of the machine. This screws fits into a small hole in the chuck. Sometimes the screw isn't well set in the hole or is well set but for some reason comes loose and causes the chuck to wobble. For some reason, this causes your problem. Now I make sure that everything is tightly secured, and I rarely have this problem. Good luck, Adam On Thu, Aug 27, 2009 at 4:47 PM, Nathan Cramer natec...@gmail.com wrote: Thanks, Robyn... I'll make sure the holder is clamped down. (sometimes its the little things...) Best Nate R J VAZQUEZ wrote: Nathan, It sounds like the blade holder is not secure enough or even in the tightness on each side. Hope this helps. Robyn Date: Thu, 27 Aug 2009 16:44:11 -0400 From: [1]natec...@gmail.com To: [2]histo...@lists.utsouthwestern.edu Subject: [Histonet] uneven alternating sections on cryostat When cutting PFA fixed, cryoprotected tissue on our cryostat, I frequently find that every other section gets cut improperly. I'll get one nicely cut section and then on the next pass I only get half of a section. This cycle simply repeats over and over and I lose many slices. It has been a while since our cryostat has been serviced, so I'm wondering if this an operator error or a machine problem. I thought maybe the tissue temperature hadn't settled properly, but the uneven cutting still happens even when I let the tissue sit for 30 minutes in the chuck holder. (cutting mouse spinal cord at -20C) On another note, if anyone has any tips for improving white/gray matter contrast in frozen spinal cord sections stained with luxol fast blue, I'd be very appreciative. I do defat the slices with chloroform and differentiate with lithium carbonate but everything is usually either very dark or very light. I am learning as I go (checking the archives here often) so any help would be great. Thanks! Nathan Cramer Neurobiology and Behavior Cornell University Ithaca, NY 14853 ___ Histonet mailing list [3]histo...@lists.utsouthwestern.edu [4]http://lists.utsouthwestern.edu/mailman/listinfo/histonet References 1. mailto:natec...@gmail.com 2. mailto:histonet@lists.utsouthwestern.edu 3. mailto:Histonet@lists.utsouthwestern.edu 4. 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
Re: [Histonet] uneven alternating sections on cryostat
Thanks, Joe and Adam. I'm always paranoid about over tightening and warping some piece of the equipment but maybe I'm being a little too gentle. I'll make sure everything is nice and snug and see how it goes. Happy slicing... Nate Galbraith, Joe wrote: Nathan: As indicated by others - first check that everything is tight as that is most often the issue - don't forget to check the adhesion of the specimen to the chuck as well. Also check the angle of approach to the blade as it could be too steep or too shallow. If everything is tight and properly adjusted, then you may have a retraction issue. If your cryostat retracts during the upstroke, the head may still be returning to the proper distance from the head to the specimen during the downstroke and hence you may see no section at first and then a gradually increasing thickness of section during the remainder of the downstroke. This can be due to two causes: 1) You can make this happen by operating the microtome at too high of a speed, if you slow down and the process improves then speed may be a factor; 2) Your microtome may be sticky and require maintenance (defrosting, cleaning, lubrication, etc.). The latter is best left to someone with extensive experience or a service rep. Good luck. Joe Galbraith University of Iowa [1]joseph-galbra...@uiowa.edu -Original Message- From: [2]histonet-boun...@lists.utsouthwestern.edu [[3]mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Adam . Sent: Thursday, August 27, 2009 4:53 PM To: Nathan Cramer Cc: [4]histo...@lists.utsouthwestern.edu Subject: Re: [Histonet] uneven alternating sections on cryostat I was having this problem too, but I think I finally figured it out. There is a screw on our cryostat that attaches the chuck to the rest of the machine. This screws fits into a small hole in the chuck. Sometimes the screw isn't well set in the hole or is well set but for some reason comes loose and causes the chuck to wobble. For some reason, this causes your problem. Now I make sure that everything is tightly secured, and I rarely have this problem. Good luck, Adam On Thu, Aug 27, 2009 at 4:47 PM, Nathan Cramer [5]natec...@gmail.com wrote: Thanks, Robyn... I'll make sure the holder is clamped down. (sometimes its the little things...) Best Nate R J VAZQUEZ wrote: Nathan, It sounds like the blade holder is not secure enough or even in the tightness on each side. Hope this helps. Robyn Date: Thu, 27 Aug 2009 16:44:11 -0400 From: [[6]1]natec...@gmail.com To: [[7]2]histo...@lists.utsouthwestern.edu Subject: [Histonet] uneven alternating sections on cryostat When cutting PFA fixed, cryoprotected tissue on our cryostat, I frequently find that every other section gets cut improperly. I'll get one nicely cut section and then on the next pass I only get half of a section. This cycle simply repeats over and over and I lose many slices. It has been a while since our cryostat has been serviced, so I'm wondering if this an operator error or a machine problem. I thought maybe the tissue temperature hadn't settled properly, but the uneven cutting still happens even when I let the tissue sit for 30 minutes in the chuck holder. (cutting mouse spinal cord at -20C) On another note, if anyone has any tips for improving white/gray matter contrast in frozen spinal cord sections stained with luxol fast blue, I'd be very appreciative. I do defat the slices with chloroform and differentiate with lithium carbonate but everything is usually either very dark or very light. I am learning as I go (checking the archives here often) so any help would be great. Thanks! Nathan Cramer Neurobiology and Behavior Cornell University Ithaca, NY 14853 ___ Histonet mailing list [[8]3]histo...@lists.utsouthwestern.edu [4][9]http://lists.utsouthwestern.edu/mailman/listinfo/histonet References 1. [10]mailto:natec...@gmail.com 2. [11]mailto:histonet@lists.utsouthwestern.edu 3. [12]mailto:Histonet@lists.utsouthwestern.edu 4. [13]http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list [14]histo...@lists.utsouthwestern.edu [15]http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list [16]histo...@lists.utsouthwestern.edu [17]http://lists.utsouthwestern.edu/mailman/listinfo/histonet References 1. mailto:joseph-galbra...@uiowa.edu 2. mailto:histonet-boun...@lists.utsouthwestern.edu 3. mailto:histonet-boun...@lists.utsouthwestern.edu 4.
RE: [Histonet] Special stains and microwave
Hi Rosa, Excuse me if I am wrong, but I think Bill is referring to using a microwave for special stains, and you are referring to a Microwave Tissue Processor. I have used an oven set at a higher temperature (85 -110 degrees C) for staining the silver portion of the GMS, and it generally only takes between 10-15 minutes. That is if you place the silver solution in the oven for approximately 10 minutes prior to use. I also use the oven set at 60-62 degrees C. for the bouins portion of the Trichrome stain. Although using a water bath at 60 degrees C. is the preferred method. You have more control using a oven than using a microwave. With a microwave, results vary with the # of slides placed in the coplin jar, as well as the placement in the microwave. There are variations in microwave intensities if you don't have a carousel. You also may have boil over of solutions. Food for thought. Akemi Akemi Allison-Tacha BS, HT(ASCP)HTL Histology Manager APMG Laboratories 105A Cooper Court, Los Gatos, CA 95032 Contact: 800.848.2764 V/M: 408.884.2718 Fax: 408.884.2758 Cell: 408.335.9994 (W) E-Mail: aallison-ta...@apmglab.com (P) E-Mail: akemiat3...@yahoo.com --- On Thu, 8/27/09, Rosa Fields rfie...@gidocs.net wrote: From: Rosa Fields rfie...@gidocs.net Subject: RE: [Histonet] Special stains and microwave To: O'Donnell, Bill billodonn...@catholichealth.net, histonet@lists.utsouthwestern.edu Date: Thursday, August 27, 2009, 2:26 PM Bill, I can highly recommend the EBS processor microwave. At a base price of 8,900 (last I checked) it is one of the lowest cost processors on the market. I don't know if having the flexibility to process in this new microwave may help justify the price? I purchased mine from Stat Lab, and the customer service from the folks at EBS was outstanding; they even had a prob and air agitator block style holder designed and built for me when I suggested that it should come with one. I really like our model, and it offers great flexibility in our lab. Rosa Fields, HT (ASCP) Gastroenterology Specialties Histology Supervisor 4545 R Street Lincoln, NE 68503 402-465-4545 rfie...@gidocs.net The information contained in the message and the documents accompanying this message contain information that is privileged and confidential and is intended only for the use of the individual or entity named above. If the reader of this message is 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 communication, other than its return to the sender, is strictly prohibited. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of O'Donnell, Bill Sent: Thursday, August 27, 2009 4:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Special stains and microwave Greetings Histonetters, We have finally come to the point where we need to fish or cut bait in relation to purchasing a laboratory approved microwave and venting it. While I think it would be cool to have one, I'm wondering if the cost is justified to do a handful of special stains that could be done (though more slowly) in a laboratory oven like the one we already have. Anyone with the time or desire to opine, please chime in. Those who have purchased such a microwave, I'll gladly take suggestions. Thanks in advance William (Bill) O'Donnell, HT (ASCP) QIHC Lead Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
