Re: [Histonet] number of slides
Here is our basic microtomy protocol: BMT - 3 HEs, each has a ribbon, plus PAS, Retic, Perls Liver bx - 3 HEs, each has a ribbon plus Retic, Trichrome, Perls, Renal bx - 4 HEs, each has a short ribbon plus PAS, PMS, trichrome - on slides with gloms Breast bx - 3 HEs, each has a ribbon Derm bx - 4 HEs, each has a ribbon, one Path likes AP PAS on all punch bx's GIT bx - 3 HEs, each has a ribbon, plus HP all other small bx's get 3 HEs, each with a ribbon and then we wait for the orders for levels and deepers and..and...and AbuDhabiAnnie On 25 March 2010 00:19, hymclab hymclab.hymc...@ministryhealth.org wrote: We follow the same practice as Susan. Our Pathologists would rather have more than less!!! Dawn -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto: histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sue Sent: Tuesday, March 23, 2010 5:08 PM To: anita dudley Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] number of slides We do three levels on all diagnostic biopsies. Liver and Kidney also get upfront special stains, as do gastric biopsies (h pylori) We did cut down on extra unstained slides since we were discarding most of them. As far as stopping levels, my pathologist thinks it goes against standard of care. As for prostate biopsies, they are so small any more, that we are thinking o cutting 4 slides staining 1and 4 for HE and holding 2 and 3 for possible IHC. We are finding that when we have to go back there is minimal tumor for IHC demonstration. Susan T. Paturzo Thomas Jefferson University Hospital ___ 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 -- Anne van Binsbergen (Hope) Abu Dhabi UAE ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] LOOKING FOR IHC POSITION
Hi, I am an HTL(ASCP) certified Histotechnologist with many years of experience. I am looking for a new position as IHC Specialist/IHC Supervisor/IHC Tech. Open to relocation. Isaac. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] H E QC
Hi there, HE QC should be carry out on every slides that are stained, if you are checking for the staining intensity, this will be carried out using a control slide on new batch of staining solution, commercial or house made. Batch number should be recorded, and slides labelled with batch reference, date and file accordingly, the same goes for daily QC of routine work. Prior each morning run a control slide is run QC. HE QC goes beyond staining intensity, as a number of artefacts can be introduced and HE unacceptable ( Shatters, creases, folds, scores, sections cut too thick, scams ... to name fews ) and therefore each slides stained should be QC by experience Staff prior been sent out to pathologist, this QC should be recorded and audited on a regular basis. Furthermore over here in the UK every laboratory belong to the UK-NEQAS http://www.ukneqas.org.uk/ and external body that run a number of EQA Schemes for histopathology. For HE EQA for each run the NEQAS will ask to select slides a number of HE slides that were produce at a randomly selected date. Slide will then be send externally and score by external accessor, and benchmark against all the participant in the scheme. If more score too low, lab will be flag and investigated. For those of you in the US, I would be curious to know if you have a similar system, and how they work. Cheers, Malika On 25 Mar 2010, at 03:22, WILLIAM DESALVO wrote: Whether you are using an automated stainer or hand staining, run a control slide and review before any patient samples are stained. I also suggest that only start or endpoint QC is not enough and you should consider incorporating continuous QC/QA at regular intervals for the stain set-up, to ensure the highest quality and provide adequate control of the process. You should be able to determine, in a very short time, the end point of the stain set up and then add QC checks for slide quality at 1/3 and 2/3 through the run or anytime a solution container is changed or rotated. In our lab, with the regents used and staining protocols available to select, we have determined that a stain set of solutions, on our automated instrument, will maintain agreed and desired quality the pathologist will accept for 1500 slides (I strongly suggest counting slides, not runs or racks). We stop processing patient slides and run the control slide at runs 1, 500, 1000 (+- 10% to allow for process flow and variance). The slides are reviewed for acceptance or rejection by a Coordinator or higher and when acceptable,patient slides may be placed on the instrument. All QC slides are saved for review and the QC maintenance sheet is filed daily. This process captures the employee that set up and monitors the instrument and the employee that QC'd along w/ the QC review results. The control slide is a multi-tissue slide that must contain the four highest volume tissue types for the lab. Each pathologist receives a daily Quality Review sheet to report any variance, issues or problems for all cases read. Think through your process, communicate with the pathologist and develop a QC/QA system/process that creates accountability for all employees, supports production of quality results and meets your regulatory needs. William DeSalvo, B.S., HTL(ASCP) System Production Manager Sonora Quest Laboratories NSH Quality Control Committee Chairperson From: adesupo2...@hotmail.com To: histonet@lists.utsouthwestern.edu Date: Wed, 24 Mar 2010 17:53:12 -0400 Subject: [Histonet] H E QC Hi, I will appreciate it, if you guys could share your method/procedure for H E QC with me. Thanking you all for your usual cooperation. Adesupo A. _ Hotmail has tools for the New Busy. Search, chat and e-mail from your inbox. http://www.windowslive.com/campaign/thenewbusy?ocid=PID27925::T:WLMTAGL:ON:WL:en-US:WM_HMP:032010_1___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _ Hotmail has tools for the New Busy. Search, chat and e-mail from your inbox. http://www.windowslive.com/campaign/thenewbusy?ocid=PID27925::T:WLMTAGL:ON:WL:en-US:WM_HMP:032010_1___ 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] Symphony
I was wondering who out there in Histo Land is using or has used the Ventana Symphony HE stainer and what their experiences have been. Thanks in advance!! Kelly D. Boyd, BS, HTL (ASCP) Lab Manager Harris Histology Services Tele (252)-830-6866 (800)-284-0672 Cell (252)-943-9527 Fax (252)-830-0032 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Job Opportunity
We have a great full time permanent opportunity for an experienced histotech at Abbott Laboratories in Abbott Park, IL. We are about 40 miles north of Chicago in Lake County. Please go to www.Abbott.com if you are interested in applying for this position. Additionally, if you would like to forward your resume to me, it could be helpful. Jackie O' ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] A.O TP8000 Processor
Hi - I'm cleaning out my office and found the procedure manual, timing disc and notes for the old A/O TP8000 Processor.If for some reason someone would like these I will mail them to you. Otherwise they are going in the trash. Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Silver Lips and Fingers
Beauty mark - like Cindy Crawford has. Andrea Grantham, HT (ASCP) Senior Research Specialist University of Arizona Cell Biology and Anatomy Histology Service Laboratory P.O.Box 245044 Tucson, AZ 85724 algra...@email.arizona.edu Tel: 520.626.4415 Fax: 520.626.2097 happy slicing and dicing and may all your stains work perfectly - Paula Sicurello P Please consider the environment before printing this email. On Mar 23, 2010, at 9:14 AM, Breeden, Sara wrote: I'm not a chemist and may shoot myself in the foot here, but if gold chloride tones silver, would it work on skin? Then you could call it a beauty spot? Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 ___ 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] Troubleshooting IHC
Hi all, I've recently run into a problem troubleshooting IHC on mouse bones. I am using the tyramide amplification system using a goat primary, anti-goat biotin, strepavidin-HRP, biotinyl tyramide, followed by another strepavidin-HRP and then DAB+ from Dako. When I use an isotype goat IgG, I get essentially no staining. I titered it once for immunofluorescence (SA-fluor instead of HRP in the last step) and got a titer of 1:800, and these staining conditions are quite reproducible. Then I titered for IHC, and 1:200 gave the best signal to noise. I did a batch of staining using those IHC conditions, and it stained beautifully, comparable to my IF. I then tried to repeat it on a second batch of sections processed in the same way, and the background was terrible (but not on my isotype slide). I thought maybe it was a problem in processing so I did a third batch of sections, and the background was still really bad. I see real staining sometimes, but I need to quantify this staining using histomorphometry, so I really need clean staining. Any ideas? The only thing I can think of is that 1:200 is just at the limit of titration that gives too much background. Thanks, Adam ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Benchmark Ultra
Hello, We currently use a Benchmark Ultra and it is worth several XTs. There are some features about it that you should know about before you start a run. What we currently do in our lab is load all of the antibodies we are likely to run every day. This can be troublesome if you run the same 30 antibodies every day (unless you have more than one Ultra, then it is much easier). We currently only run 30 total antibodies on our Ventana platform and of those 30 we run 10 or 12 every day. When we come in, we pull a list of stains for the AM run off the computer and we have a good idea if we need to load something less common along with the list of 12 more common antibodies and a detection kit. For the next run, if we have already started a run and we need to add an antibody, we will be forced to use the Landing Zone feature. Using this, we are able to load the missing antibodies/reagents and remove what we don't need. With regard to the important or unimportant cases, you can flag them with the blue LED, (which looks really cool, by the way...) so that you know visually which cases you are looking for without having to hover over a position on the computer. As far as loading important cases after unimportant cases, I don't think I can offer any advice there. The instrument is essentially 30 individual stainers and you can choose whether or not to load the slides. We don't really encounter this issue because of the way we have structured our workflow, but then again, we have enhanced our workflow based upon the capabilities of the instrumentation that we use. I hope this has helped. If there are any other questions, feel free to email me. Ashley Troutman BS, HT(ASCP) QIHC Vanderbilt University Histopathology 1301 Medical Center Drive TVC 4532 Nashville, TN 37232 Date: Wed, 24 Mar 2010 19:37:25 +0100 From: Gudrun Lang gu.l...@gmx.at Subject: [Histonet] benchmark ultra and continuous workflow To: histonet@lists.utsouthwestern.edu Message-ID: b646f3f46af8408cafc5b4887d948...@dielangs.at Content-Type: text/plain; charset=us-ascii Hi Benchmark Ultra users! I would like to hear of your experiences with the continuous workflow of this instrument. Does it really make life easier or even more complicated? I think of handling one slide every few minutes after the staining is completed and of the problems, that occur when the stainer is started with unimportant cases and the later coming important cases have not enough place to complete them in time. Gudrun Lang ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: chick embryo frozen sections
hello, will some one please give me pointers on successful methods used for cryosectioning bursa esophagus from approx. 21 day chick embryos (such as recommended temp. micron thickness, for yielding the best sections)? Thanks! Atoska ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Acetone Fixation
hello, if any of you have acetone fixation incorporated into you frozen section H E staining protocol will you please advise me on adjustments necessary for routine staining protocol. Also, out of curiosity please enlighten me on the purpose for post sectioning acetone fixation on tissue samples initially fixed in 95% ETOH/ Acetic Acid? Your prompt replies will be greatly appreciated. ~ Atoska ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: coverslipper
It depends on your volume and how long you maintain your slides. Because you are a children's hospital you keep them longer. The Leica brand is very troublesome in high volume situations. I personally prefer Sakura tape for the speed and fast drying time. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rathborne, Toni Sent: Thursday, March 25, 2010 11:15 AM To: Margaryan, Naira; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] coverslipper Leica CV5030. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu]on Behalf Of Margaryan, Naira Sent: Thursday, March 25, 2010 12:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] coverslipper Hi Colleges, I need your opinion about coverslip by hand vs. using machine. If you use machine what company's coverslipper you prefer? Thanks in advance, Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Be sure to visit Somerset Medical Center's Web site - www.somersetmedicalcenter.com - for the most up-to-date news, event listings, health information and more. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: The information in this e-mail may be confidential and/or privileged. If you are not the intended recipient or an authorized representative of the intended recipient, you are hereby notified that any review, dissemination, or copying of this e-mail and its attachments, if any, or the information contained herein is prohibited. If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail from your computer system. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] coverslipper
We've had our Sakura film coverslipper since '94 and love it. The goodsaves us tech time coverslipping for hours each day, plus you can file the slides right away. The bad.the docs prefer the bone marrow smears to be coverslipped by hand. We have about 12 bone marrows a monthso reallyit's not a problem. We've also tried other brands of film, but the cytotechs could notice a refractive difference, so we switched back to the Sakura brand. Margaryan, Naira nmargar...@childrensmemorial.org 3/25/2010 12:07 PM Hi Colleges, I need your opinion about coverslip by hand vs. using machine. If you use machine what company's coverslipper you prefer? Thanks in advance, Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Acetone Fixation
What is your goal? A routine HE or immuno'setc.?? Atoska Gentry gent...@auburn.edu 3/25/2010 12:21 PM hello, if any of you have acetone fixation incorporated into you frozen section H E staining protocol will you please advise me on adjustments necessary for routine staining protocol. Also, out of curiosity please enlighten me on the purpose for post sectioning acetone fixation on tissue samples initially fixed in 95% ETOH/ Acetic Acid? Your prompt replies will be greatly appreciated. ~ Atoska ___ 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] coverslipper
Leica CV3050 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Margaryan, Naira Sent: Thursday, March 25, 2010 12:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] coverslipper Hi Colleges, I need your opinion about coverslip by hand vs. using machine. If you use machine what company's coverslipper you prefer? Thanks in advance, Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] whole mount protocol
hi all i am trying to do an in situ hybridization in whole mount brains at P7 but is hard to get staining mostly due to permeabilization issues since is 7 days postnatal brain. I would really appreciate to receive any suggestion or protocol to help me in this issue. Thanks a lot in advance Carlos Carlos G. Perez-Garcia, Ph.D. Molecular Neurobiology Lab (MNL-O) The Salk Institute 10010 North Torrey Pines Road 92037 La Jolla, CA, USA Phone: 858-453-4100 ext. X1449 Fax: 858 558 6207 E-mail: cpgar...@salk.edu  ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: coverslipper
Thanks a lot to all of you answered me. I was surprise nobody mentioned coverslipper from DAKO. Are any of you have any experience with DAKO's coverslipper? Again Thanks to all, Naira Subject: coverslipper Hi Colleges, I need your opinion about coverslip by hand vs. using machine. If you use machine what company's coverslipper you prefer? Thanks in advance, Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Histonet Digest, Vol 76, Issue 38
On 3/25/10 1:04 PM, histonet-requ...@lists.utsouthwestern.edu histonet-requ...@lists.utsouthwestern.edu wrote: Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than Re: Contents of Histonet digest... Today's Topics: 1. Re: H E QC (Sue) 2. RE: H E QC (WILLIAM DESALVO) 3. Re: number of slides (Anne van Binsbergen) 4. LOOKING FOR IHC POSITION (Isaac O) 5. Re: H E QC (Malika Benatti) 6. RE: LOOKING FOR IHC POSITION (Plewinski, Amy) 7. Symphony (Kelly Boyd) 8. Job Opportunity (Jackie M O'Connor) 9. A.O TP8000 Processor (Cheryl Crowder) 10. RE: Symphony (CHRISTIE GOWAN) 11. Re: Silver Lips and Fingers (Andrea Grantham) 12. Troubleshooting IHC (Adam .) 13. Benchmark Ultra (Troutman, Kenneth A) 14. coverslipper (Margaryan, Naira) 15. RE: coverslipper (Rathborne, Toni) 16. Re: chick embryo frozen sections (Atoska Gentry) 17. Re: Acetone Fixation (Atoska Gentry) 18. RE: coverslipper (Nails, Felton) -- Message: 1 Date: Wed, 24 Mar 2010 23:02:35 + (UTC) From: Sue suetp...@comcast.net Subject: Re: [Histonet] H E QC To: ADESUPO ADESUYI adesupo2...@hotmail.com Cc: histonet@lists.utsouthwestern.edu Message-ID: 1021401464.930521269471755920.javamail.r...@sz0028a.westchester.pa.mail.comca st.net Content-Type: text/plain; charset=utf-8 We have automatic stainers, so after the stainer is set up a slide with a micro-array is run. This slide is checked by the histologists and logged in. The next slides run are our rapid cases. A log sheet is prepared and handed to the pathologist with the slides and they are graded for processing, embedding, microtomy and staining. This log is turned into the supervisor and reviewed daily. If there are issues the supervisor will review with the histologists. Since the techs rotate weekly we are able to monitor all the tech's technical performance. Susan T. Paturzo TJUH -- Message: 2 Date: Wed, 24 Mar 2010 21:22:34 -0600 From: WILLIAM DESALVO wdesalvo@hotmail.com Subject: RE: [Histonet] H E QC To: adesupo2...@hotmail.com, histonet histonet@lists.utsouthwestern.edu Message-ID: blu103-w2204b7c6ba7d8c137df3f491...@phx.gbl Content-Type: text/plain; charset=iso-8859-1 Whether you are using an automated stainer or hand staining, run a control slide and review before any patient samples are stained. I also suggest that only start or endpoint QC is not enough and you should consider incorporating continuous QC/QA at regular intervals for the stain set-up, to ensure the highest quality and provide adequate control of the process. You should be able to determine, in a very short time, the end point of the stain set up and then add QC checks for slide quality at 1/3 and 2/3 through the run or anytime a solution container is changed or rotated. In our lab, with the regents used and staining protocols available to select, we have determined that a stain set of solutions, on our automated instrument, will maintain agreed and desired quality the pathologist will accept for 1500 slides (I strongly suggest counting slides, not runs or racks). We stop processing patient slides and run the control slide at runs 1, 500, 1000 (+- 10% to allow for process flow and variance). The slides are reviewed for acceptance or rejection by a Coordinator or higher and when acceptable,patient slides may be placed on the instrument. All QC slides are saved for review and the QC maintenance sheet is filed daily. This process captures the employee that set up and monitors the instrument and the employee that QC'd along w/ the QC review results. The control slide is a multi-tissue slide that must contain the four highest volume tissue types for the lab. Each pathologist receives a daily Quality Review sheet to report any variance, issues or problems for all cases read. Think through your process, communicate with the pathologist and develop a QC/QA system/process that creates accountability for all employees, supports production of quality results and meets your regulatory needs. William DeSalvo, B.S., HTL(ASCP) System Production Manager Sonora Quest Laboratories NSH Quality Control Committee Chairperson From: adesupo2...@hotmail.com To: histonet@lists.utsouthwestern.edu Date: Wed, 24 Mar 2010 17:53:12 -0400 Subject: [Histonet] H E QC Hi, I will appreciate it, if you guys could share your method/procedure for H E QC
Re: [Histonet] coverslipper
We use the Leica CV5030 the advantage of it is that it can be attached to their Auto stainer providing continuous workflow as rack and coverslip 30 slides per run, without the need to physically transfer slides rack between the autostainer and coverslipper or used as a stand alone coverslipper if needed, though at time it can be problematic. For a start glass cover slip must be kept at 37 oC prior use otherwise they tend to stick to each others. Also does not recognised all the slides type unless they are Leica one, or the coverlslipper as been calibrated to use a specific slides type. In the past I used the Sakura Acetate film coverslipper, and their were no major problem with it though remounting section was a very tedious process. Malika On Thu, Mar 25, 2010 at 5:37 PM, Lynette Pavelich lpave...@hurleymc.comwrote: We've had our Sakura film coverslipper since '94 and love it. The goodsaves us tech time coverslipping for hours each day, plus you can file the slides right away. The bad.the docs prefer the bone marrow smears to be coverslipped by hand. We have about 12 bone marrows a monthso reallyit's not a problem. We've also tried other brands of film, but the cytotechs could notice a refractive difference, so we switched back to the Sakura brand. Margaryan, Naira nmargar...@childrensmemorial.org 3/25/2010 12:07 PM Hi Colleges, I need your opinion about coverslip by hand vs. using machine. If you use machine what company's coverslipper you prefer? Thanks in advance, Naira ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Smile it confuses people ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] coverslipper
I purchased a used Hacker RCM-3660 glass coverslipper several years ago. It was missing some parts and the nice people from Hacker supplied me with the needed parts. This work horse has been running 7 days a week for the past several years and has never given us any problems. Great machine and great people. Jim ___ James E. Staruk HT(ASCP) www.masshistology.com www.nehorselabs.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Malika Benatti Sent: Thursday, March 25, 2010 3:08 PM To: Lynette Pavelich Cc: histonet@lists.utsouthwestern.edu; Naira Margaryan Subject: Re: [Histonet] coverslipper We use the Leica CV5030 the advantage of it is that it can be attached to their Auto stainer providing continuous workflow as rack and coverslip 30 slides per run, without the need to physically transfer slides rack between the autostainer and coverslipper or used as a stand alone coverslipper if needed, though at time it can be problematic. For a start glass cover slip must be kept at 37 oC prior use otherwise they tend to stick to each others. Also does not recognised all the slides type unless they are Leica one, or the coverlslipper as been calibrated to use a specific slides type. In the past I used the Sakura Acetate film coverslipper, and their were no major problem with it though remounting section was a very tedious process. Malika ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] coverslipper
Is this coverslipper now produced by Medite -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of jstaruk Sent: Thursday, March 25, 2010 2:40 PM To: malbena...@gmail.com; 'Lynette Pavelich' Cc: histonet@lists.utsouthwestern.edu; 'Naira Margaryan' Subject: RE: [Histonet] coverslipper I purchased a used Hacker RCM-3660 glass coverslipper several years ago. It was missing some parts and the nice people from Hacker supplied me with the needed parts. This work horse has been running 7 days a week for the past several years and has never given us any problems. Great machine and great people. Jim ___ James E. Staruk HT(ASCP) www.masshistology.com www.nehorselabs.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Malika Benatti Sent: Thursday, March 25, 2010 3:08 PM To: Lynette Pavelich Cc: histonet@lists.utsouthwestern.edu; Naira Margaryan Subject: Re: [Histonet] coverslipper We use the Leica CV5030 the advantage of it is that it can be attached to their Auto stainer providing continuous workflow as rack and coverslip 30 slides per run, without the need to physically transfer slides rack between the autostainer and coverslipper or used as a stand alone coverslipper if needed, though at time it can be problematic. For a start glass cover slip must be kept at 37 oC prior use otherwise they tend to stick to each others. Also does not recognised all the slides type unless they are Leica one, or the coverlslipper as been calibrated to use a specific slides type. In the past I used the Sakura Acetate film coverslipper, and their were no major problem with it though remounting section was a very tedious process. Malika ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: The information in this e-mail may be confidential and/or privileged. If you are not the intended recipient or an authorized representative of the intended recipient, you are hereby notified that any review, dissemination, or copying of this e-mail and its attachments, if any, or the information contained herein is prohibited. If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail from your computer system. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Troubleshooting IHC
Yes. I block in 3% H2O2, followed by protein block (it's a mysterious buffer called TNB that comes with the tyramide amplification kit), and then avidin/biotin. Adam On Thu, Mar 25, 2010 at 2:25 PM, Margaryan, Naira nmargar...@childrensmemorial.org wrote: Hi Adam, How do you block? I usually have: H2O2, Avidin/Biotin and Protein blocking steps. Naira Message: 12 Date: Thu, 25 Mar 2010 10:43:28 -0500 From: Adam . anonwu...@gmail.com Subject: [Histonet] Troubleshooting IHC To: histonet@lists.utsouthwestern.edu Message-ID: 858249121003250843h2d0f16a2q1a74ad1091630...@mail.gmail.com Content-Type: text/plain; charset=ISO-8859-1 Hi all, I've recently run into a problem troubleshooting IHC on mouse bones. I am using the tyramide amplification system using a goat primary, anti-goat biotin, strepavidin-HRP, biotinyl tyramide, followed by another strepavidin-HRP and then DAB+ from Dako. When I use an isotype goat IgG, I get essentially no staining. I titered it once for immunofluorescence (SA-fluor instead of HRP in the last step) and got a titer of 1:800, and these staining conditions are quite reproducible. Then I titered for IHC, and 1:200 gave the best signal to noise. I did a batch of staining using those IHC conditions, and it stained beautifully, comparable to my IF. I then tried to repeat it on a second batch of sections processed in the same way, and the background was terrible (but not on my isotype slide). I thought maybe it was a problem in processing so I did a third batch of sections, and the background was still really bad. I see real staining sometimes, but I need to quantify this staining using histomorphometry, so I really need clean staining. Any ideas? The only thing I can think of is that 1:200 is just at the limit of titration that gives too much background. Thanks, Adam ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Direction for TP 800
Hi - Got a taker for the directions. Who know? Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] primate brain
I have some primate brain blocks to do. When I section the blocks the sections look nice. When I put the ribbon on the water bath( temp is 47) the tissue is wrinkled around the edges. No amount of time on the water makes this better. Ideas to solve this problem would be appreciated. Thanks. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] primate brain
At what thickness do you cut your sections ? For Human brain we cut section at room temperature moist in Molifex and cut sections at 7 µm for standard HE/ 14 µm for LFB. Not sure what is the melting point of the wax your use, but if it is around 57 oC you can safely raise the temperature of your water bath to 50 oC. Hope this help. Malika On Thu, Mar 25, 2010 at 9:17 PM, Beth A Gray g...@rarc.wisc.edu wrote: I have some primate brain blocks to do. When I section the blocks the sections look nice. When I put the ribbon on the water bath( temp is 47) the tissue is wrinkled around the edges. No amount of time on the water makes this better. Ideas to solve this problem would be appreciated. Thanks. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Smile it confuses people ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
