Re: [Histonet] Validating a new manual special stain
The new CAP Checklist (published 6/4/2020) no longer requires validations for "/LDTs that employ the following methods: manual microscopy eg. histopathologic and cytopathologic microscopic interpretation/" (see COM.40350, note 8). So, CAP does not require it and won't have guidance for that. HOWEVER, CLIA still requires validations! The common way to do that is to run a minimum of 5, and preferably 10, different samples by the new method and grade them according to expected results. A few of the samples should be 'negative' samples. Using various control tissue for your validations is perfectly acceptable. Write that up, have the medical director review the slides and sign and date the bottom of the form: "This procedure has been validated and is approved for patient use." Beth Cox, HTL/SCT(ASCP)QIHC Pathology Solutions Prescott, MI -- Message: 1 Date: Fri, 6 Nov 2020 18:35:25 + From: "Pairan, Kelly" To: "'histonet@lists.utsouthwestern.edu'" Subject: [Histonet] Validating a new manual special stain Message-ID: <49e4f7123d03401385e3727519640...@l1perdwmbx04.childrensroot.net> Content-Type: text/plain; charset="us-ascii" Good Afternoon, We are currently updating one of our manual special stain procedures. How have you documented the validating of a new special stain method at your institution? We have a IHC validation protocol that follows the CAP guidelines but the CAP checklist does not mention special stain validations. Thanks for your input, Kelly -- Subject: Digest Footer ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- End of Histonet Digest, Vol 204, Issue 4 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cytology (Special stains & IHC)
Hi Karen, The best resource for Cyto questions would probably be a recent edition of the Carson Histotechnology book. The Fourth edition has some info on special stains for Cyto, but the new Fifth edition is expanded on that topic and has more info on IHCs for Cyto (Chapter 17). For _special stains_, you would definitely want them fixed in 95% alcohol. The only exception to that would be fat stains which would of course require air-dried slides. It would likely be advantageous to use charged slides, but that is not required. Remember when special staining Cyto slides fixed in 95% alcohol, that the deparaffinization steps at the beginning of the procedure must be skipped. You are normally okay with using regular FFPE controls with your Cytospins or ThinPrep special stains. Doing _IHCs_ on Cytology preps (Cytospin or ThinPrep) gets a little trickier. _IF_ you are going to do them, you would want them to be 95% alcohol fixed and on charged slides. However, you would be required to do a validation for Cytology for any/all antibodies you were going to run. (Your CAP or CLIA inspector will eat you alive if you don't!). Alcohol fixed specimens will require a new optimization since the pretreatments will be different than those for FFPE. Many antibodies, particularly nuclear ones, may be difficult or impossible to stain on the intact cells in a Cyto prep. Also, you would not be able to use FFPE controls for Cyto IHC since they are 'fixed and processed differently" than the Cyto slide and therefor don't properly control the process. AND, alcohol fixed smears begin to lose their antigenicity in about 24 hours, so they must be stained the same day the slides are made, so having a control bank of alcohol fixed slides isn't feasible. Doing IHC on Cyto smears using FFPE protocols and FFPE controls is very likely to give you false negative results. Unless you are doing large volume Cyto, it is very strongly recommended that you do your IHCs for Cyto on the FFPE cell block. Beth Cox, HTL/SCT(ASCP)QIHC -- Message: 5 Date: Wed, 5 Aug 2020 15:34:28 + From: "Heckford, Karen - SMMC-SF" To:"histonet@lists.utsouthwestern.edu" Subject: [Histonet] Cytology Message-ID:<903be99abb344593a73079761d762...@phx-exch-013.chw.edu> Content-Type: text/plain; charset="us-ascii" Good Morning, I rarely do Special stains or IHC's on Cytology cytospins or thin prep type slides.Should these slides be charged, air dried or fixed in 95% alcohol before doing Special stains or IHC's on them? Is there a good book or some sort of publication on cytology procedures regarding the above mentioned. I have tried and look it up and it seems all over the place on what to do. Any help would be greatly appreciated, Karen Heckford HT ASCP CE Lead Histology Technician St. Mary's Medical Center 450 Stanyan St. San Francisco, Ca. 94117 415-668-1000 ext. 6167 karen.heckf...@dignityhealth.org<mailto:karen.heckf...@dignityhealth.org> ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] plural fluid prep
Hi Michelle, When I was in my Cytology training, we did an experiment to answer just that question. We started with a 2 liter jug of plural fluid with nothing added, and we stored in in the fridge. At first we tested it every day to see if there was any degeneration, then we got tired of that and tested twice a week, then once a week. After 4 months it was still just fine, and we threw it away because we were tired of testing it (and it was the end of the semester!). It seems that the cells are happy in their natural habitat and refrigeration slowed/prevented any bacterial growth. So my advice is: don't add anything to it, keep it in the fridge, and it will last many months!! Beth Cox, SCT/HTL(ASCP)QIHC From: Michelle Jamison To:"histonet@lists.utsouthwestern.edu" Subject: [Histonet] plural fluid prep Message-ID: Content-Type: text/plain; charset=utf-8; format=flowed Hi All, I work in a research lab where we are using plural fluid to proof a method. Recently we have received some, and are unsure how long it will be ok in the fridge, or if we should add some preservation fluid to it. Can anyone advise please. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Histonet,
Lisa, I have sent you a booklet on detailed Cell Block procedures as an attachment in a separate email -- if anyone else is interested in a copy, please email me directly. Thanks Beth Cox, SCT/HTL(ASCP)QIHC -- Message: 1 Date: Wed, 25 Apr 2018 17:11:43 + From: "White, Lisa M."<lisa.whi...@va.gov> To:"histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Subject: [Histonet] Cell Block Message-ID: <bn7pr09mb2547b2b7f5d6d15ff1c58bdfd0...@bn7pr09mb2547.namprd09.prod.outlook.com> Content-Type: text/plain; charset="us-ascii" Does anyone have the recipe for making cell blocks with Thrombin and Plasma? We made them at my first Histo job, oh say over 20 years ago but I do not remember the ratio. Thanks bunches, Lisa ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] PAP staining quality
Hi Charles, A couple things to check on: 1. The first concern I would have is your EA stain. Poor EA staining will give too much orange staining and pale other counterstains (making them look aged).What brand are you using? Have you changed brands? Is your EA close to the expiration date? Is the bulk stored with light exposure? I think fixing your EA will fix all the other problems. 2. The other question I have regards your alcohol. Have you changed types/brands? Pap staining is very delicate and the different alcohols used can make a big difference. Beth Cox, HTL/SCT(ASCP)QIHC -- Message: 3 Date: Thu, 4 Feb 2016 09:50:18 -0500 From: Charles Riley <cri...@dpspa.com> To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PAP stain quality Message-ID:
Re: [Histonet] Pathology labs using Premier for staffing
The way Premier can break down the productivity/staffing info for your lab is dependent on how your organization separates the lab into cost centers -- if your lab is all one cost center for clinical and anatomic labs together, that's the way it gets reported to Premier, and that's how the statistics will come back to you. These statistics are based on billable tests and worked hours for your cost center. If your organization has the anatomic lab as a different cost center from the clinical lab, the info will get reported to Premier separately and Premier can give you info back comparing you to other AP only databases. There are no labs I am aware of which have Histo and Cyto in separate cost centers, so Premier would likely be unable to give you comparison data that way. Beth Cox, HTL/SCT(ASCP)QIHC Pathology Solutions, Inc Message: 10 Date: Thu, 15 Oct 2015 11:18:39 -0400 From: Amy Self<as...@tidelandshealth.org> To:"histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Subject: [Histonet] Pathology labs using Premier for staffing Message-ID: <d84a63d984914348a9e9a86492d86016d9fa262...@gmhdtcexch.gmhpost.com> Content-Type: text/plain; charset="us-ascii" Dear Histonetters, Are there any labs that utilize Premier for staffing productivity? For those that do use Premier is the histology section separated from your other lab sections or included with the clinical lab? And also is cytology considered separate or included with histology? Do you report volume and staffing? Or, only report one or the other? I am trying to find a comparison group for staffing productivity. Our histology and cytology sections are included as part of the entire lab for both volume and staffing. Thanks in advance for your help, Amy Self Histology Lab Senior Tech Lab Tidelands Georgetown Memorial Hospital 606 Black River Road Georgetown, SC 29440 843-520-8711 as...@tidelandshealth.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: QIHC exam
Amylin, The Michigan Society of Histotechnologists has Study Workbook for the QIHC exam. It was written to cover all the things you need to study for the exam (and avoids the things not covered on the exam). To order, go to mihisto.org cost:$20 Beth Cox, HTL/SCT(ASCP)QIHC Pathology Manager Beebe Healthcare Lewes, De Message: 3 Date: Mon, 20 Oct 2014 17:39:35 + From: Amy Johnsonajohn...@aipathology.com Subject: [Histonet] QIHC exam To:'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu, 'histonet-boun...@lists.utsouthwestern.edu' histonet-boun...@lists.utsouthwestern.edu Message-ID: D2A2F4D6FC62CB47A21B179D1812DC5212276C@SERV2012.aipathology.local Content-Type: text/plain; charset=us-ascii Is there any info or mock exams on the QIHC that people can get to see if they are ready to take the exam?? Thanks, Amylin Johnson, BS HTL(ASCP) Associates in Pathology Wausau, WI 715-847-2130 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Ventana Ultraview users
Beth, We are doing exactly that same thing right now. This is changing a complete detection system, so we need a full validation (10 pos + 10 negs for all non-prognostic markers). To avoid running 20 slides on every antibody on our menu (), we have chosen to use a 3-prong method: 1. we bought appropriate TMAs from reputable vendors (such as Pantomics, Inc) which will provide more than 10 Pos +10 neg for most cases. 2. we made a universal negative control with 12 different tissues from our collection (provides tissues processed by us) 3. we run 1(or 2) of our control slides (bonus: also validates the control at the same time) This way we only run three slides for each validation to get the required number of tissues - big difference in workload and cost! Beth Cox, HTL/SCT(ASCP)QIHC Pathology Solutions, Inc __ Message: 1 Date: Wed, 9 Jul 2014 12:16:55 -0500 From: Beth Brinegarbbrinegar...@gmail.com Subject: [Histonet] Ventana ultraview users To:histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Message-ID: CAFYmSULScGQdaSdrU0dBz=55yaq-qlwsm_z944a1hdeniw7...@mail.gmail.com Content-Type: text/plain; charset=UTF-8 Hello all, Our lab is getting ready to switch from using the iview DAB to the ultraview - any tips for validation? How many cases did you run for each antibody? Thanks! Beth Brinegar HTL(ASCP) Anatomic Pathology Supervisor Mercy Medical Center Cedar Rapids, IA 52403 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: HTL Certification
Hi Cecy, I recommend the Michigan Society of Histotech's HT/HTL Study Workbook. This will take you through everything you need for the HTL in an organized and clear manner (much better than the ASCP outline). Note that this is a workbook that you fill in, not a question/answer book. It has forms and charts to lead you through all your study. It will help you understand the info, much better than just memorizing. I think the cost is $25. you can order one at: www.mihisto.org Beth Cox, HTL/SCT(ASCP)QIHC Vagabond Histotech _ Message: 1 Date: Mon, 19 May 2014 12:32:36 -0500 From: Cecy Stancecysta...@gmail.com Subject: [Histonet] HTL certification To:histonet@lists.utsouthwestern.edu Message-ID: CACh=0wgaaEfVDFmsuB-Jw8OKo=lkupapgujjkoeagjazasx...@mail.gmail.com Content-Type: text/plain; charset=UTF-8 Hello everyone, I'm starting to prepare for my HTL certification (I am very nervous and anxious but also very excited about this decision to go for it). I was just curious to know how you guys prepared for it, and how long it took for you to prepare before taking the test. Will 6 months preparation be enough? (I know that may depend on the individual; it's just that I had my Masters over 10 years ago and I haven't studied this much since then). I have Freida L. Carson's 3rd Edition book -- quite daunting to memorize -- but the outline ASCP has provided for study seems to be helpful. Do you have any other book/study aid suggestions? Thank you in advance for your input and advice! C.A. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Anyone in Uganda??
Is there anyone on the Histonet in Uganda?? I have questions regarding licensing/certification/training for lab people there. Please feel free to contact me directly. Beth Cox, HTL/SCT(ASCP)QIHC ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Cornflaking
Sharon other Histonetters, Cornflaking is literally microscopic air trapped under the coverslip. It doesn't have anything to do with poor dehydration and trapped water. Thus it can be caused by partial drying out before the coverslipping. The things you need to look at to eradicate the problem is: 1. Keep the slides wet before coverslipping (obviously) 2. Consider increasing the amount of xylene deposited on the slide for tape coverslippers. 3. For glass coverslippers, consider increasing the xylene and increasing or changing the mounting media. Cornflaking tends to happen more on slides/sections with rough topography on the section (the more rough it is, the more nooks crannies to trap the air). So anything that would give the section a more rough surface would increase the tendency to cornflake; such as: section lifting. section thickness, even chatter in the sections. Think about things that would affect the section - for example, are you using a different brand of blade on the microtome? Hope this helps, Beth Cox, HTL/SCT(ASCP)QIHC Message: 15 Date: Thu, 13 Mar 2014 11:59:06 -0400 From: sris...@mail.holyname.org Subject: Re: [Histonet] RE: Cornflaking artifact To: HERRINGTON, SHEILA sheila.herring...@interiorhealth.ca Cc: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu,Sharon Scalise sscal...@beaumont.edu, histonet-boun...@lists.utsouthwestern.edu, 'Laurie Colbert' lcolb...@pathmdlabs.com Message-ID: of46844f54.2f7fc911-on85257c9a.00575cb9-85257c9a.0057c...@holyname.org Content-Type: text/plain; charset=US-ASCII We had this problem several years ago. We were using the sakura tapes with the coverslipper. We did the following: Last three alcohols were changes frequently. Slides should be not dry when loading on coverslipper. If you could load two racks at a time, only load one. By this way the slides in the second rack will not dry out. Finally, change the tapes from sakura to Mercedes Medical tapes. Mala Nirmala Srishan Holy Name Medical Center From: HERRINGTON, SHEILA sheila.herring...@interiorhealth.ca To: 'Laurie Colbert' lcolb...@pathmdlabs.com, Sharon Scalise sscal...@beaumont.edu, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Date: 03/13/2014 11:43 AM Subject:[Histonet] RE: Cornflaking artifact Sent by:histonet-boun...@lists.utsouthwestern.edu We also have recently started to see this artifact more than ever before, and nothing in our process has changed. We have tried everything to correct to no avail. Wonder if it is possible to be a change in some type of supply, either xylene or coverslipping film. Something has changed but am at a loss as to what. Sheila Herrington Technical Lead Histopathology and Immunohistochemistry Kelowna General Hospital 2268 Pandosy Street, Kelowna, B.C. V1Y 1T2 250-862-4300 ext 7587 or 7510 sheila.herring...@interiorhealth.ca -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [ mailto:histonet-boun...@lists.utsouthwestern.eduhistonet-boun...@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert Sent: Thursday, March 13, 2014 6:30 AM To: Sharon Scalise; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Cornflaking artifact You will also see the cornflaking if your tissue is lifting off of the slide at all. We used to get this more often on hard, decal specimens than on other specimens. We used the film to coverslip. If you remove the film from the problem slides and recoverslip conventionally with extra mountant and glass coverslips, I'm sure you will not see the artifact. Laurie Colbert -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [ mailto:histonet-boun...@lists.utsouthwestern.eduhistonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sharon Scalise Sent: Wednesday, March 12, 2014 8:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Cornflaking artifact I am looking for help with cornflaking (tiny, brown dry spots under coverslip)artifact. We have been using fresh xylene on our stainer and coverslipper, cleaned and wiped all containers dry before filling, tried different lots of coverslipping film and had service on our coverslipper to make sure it was functioning properly, including the xylene drip. We continue to have this artifact and it is driving us crazy. It is sporadic with no pattern of tissue type or placement on the slide. Sometimes it lands on tissue other times not. Most of the time when we remove the coverslip and re-coverslip it goes away (I am assuming because the acetone removes any minute amounts of water that may be present). We just cannot figure out where the water is coming from. Has anyone seen this artifact while using the drying step on the prisma stainer? We just recently started using the drying on some slides and I am thinking maybe it is causing humidity??? I cannot say for a fact that our cornflaking started at the same
[Histonet] Re: Staining on Alcohol Fixed Smears
I'm sorry, I have no experience with acetone fixed smears. Beth Cox, HTL/SCT(ASCP)QIHC Vagabond Histotech On 12/17/2013 8:40 AM, Susan Foreman wrote: What info do you guys have on acetone fixed smears? Specifically, having trouble getting PAX5 (nuclear) to show good signal without chewing up the cells during antigen retrieval. Susan Foreman, HT (ASCP) KDL Pathology 315 Erin Drive Knoxville, TN 37919 (865)584-1933 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tom McNemar Sent: Tuesday, December 17, 2013 5:48 AM To: 'Beth Cox'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Staining on Alcohol Fixed Smears Thanks for the clarification. I appreciate the info! Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcne...@lmhealth.org www.LMHealth.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Beth Cox Sent: Monday, December 16, 2013 2:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Staining on Alcohol Fixed Smears I agree that with no formalin exposure, there is no crosslinking that needs to be broken, but a then, do nothing approach to antigen retrieval oversimplifies the issue. Alcohol fixation makes changes in the protiens/antibodies also, although different changes than formalin does, and these changes need to be accounted for. In my research on alcohol fixed smears, I found that most antibodies perform best with some antigen retrieval, often gentler than formalin tissues require. Beth Cox, HTL/SCT(ASCP)QIHC Vagabond Histotech -- Message: 4 Date: Mon, 16 Dec 2013 11:13:59 -0500 From: Tom McNemartmcne...@lmhealth.org Subject: RE: [Histonet] Re: Staining on Alcohol Fixed Smears To: 'Beth Cox'bethc...@gmail.com, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu,levi.fr...@gmail.com levi.fr...@gmail.com Message-ID: E9A90E28259D2F4E84308C5E8EA8F7B4016951E94252@lmhs-exchange Content-Type: text/plain; charset=us-ascii Since there is no exposure to formalin, we do not do antigen retrieval on alcohol fixed specimens. We use the Ventana Benchmark XT and copy the protocol to a new number and use the Wet option rather than paraffin. No retrieval needed. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcne...@lmhealth.org www.LMHealth.org -Original Message- From:histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Beth Cox Sent: Saturday, December 14, 2013 6:30 PM To:histonet@lists.utsouthwestern.edu;levi.fr...@gmail.com Subject: [Histonet] Re: Staining on Alcohol Fixed Smears Levi, I've worked up most of the more common antibodies on alcohol fixed smears. There are two things to remember here: 1) your specimens are alcohol fixed, meaning antigen retrieval needs are different, and 2) they are smears, meaning the cells weren't 'cut open' during microtomy, which impeeds some nuclear staining. Those are two different issues to consider. Some rules of thumb: 1) most cell membrane antigens will stain nicely with minimal changes to the protocols you use for FFPE specimens 2) most cytokeratin antigens stain nicely with little change to your FFPE protocols (if you need to change, start by cutting your antigen retrieval time in half) 3) most nuclear antigens require significant changes from FFPE protocols, and some are essentially impossible to stain using standard antibodies. When I return to work on Monday, I will forward you my protocol 'adjustments' for the antibodies you requested. Beth Cox, HTL/SCT(ASCP)QIHC Vagabond Histotech On 12/14/2013 12:00 PM,histonet-requ...@lists.utsouthwestern.edu wrote: Message: 4 Date: Sat, 14 Dec 2013 18:25:08 +0200 From: Levi Friedlevi.fr...@gmail.com Subject: [Histonet] Staining on Alcohol Fixed Smears. To:histonet@lists.utsouthwestern.edu Message-ID: CA+puqxTXL85pEwvR=sqPQA36tMJPm=sx2kv3gyebrbu0he3...@mail.gmail.com Content-Type: text/plain; charset=ISO-8859-1 Hi Everyone. I am looking to stain alcohol fixed smears with a group of antibodies. The antibodies of particular interest are p63, p53 and ki67. Along with these antibodies I am looking for positive nuclear and cytoplastic staining antibodies. If anyone has any experience in staining alcohol fixed slides your information is very appreciated. All the best. -- Sincerely, - Levi Fried ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, including attachments, is intended for the sole use of the individual and/or entity to whom it is addressed, and contains information from Licking
[Histonet] Re: Staining on Alcohol Fixed Smears
I agree that with no formalin exposure, there is no crosslinking that needs to be broken, but a then, do nothing approach to antigen retrieval oversimplifies the issue. Alcohol fixation makes changes in the protiens/antibodies also, although different changes than formalin does, and these changes need to be accounted for. In my research on alcohol fixed smears, I found that most antibodies perform best with some antigen retrieval, often gentler than formalin tissues require. Beth Cox, HTL/SCT(ASCP)QIHC Vagabond Histotech -- Message: 4 Date: Mon, 16 Dec 2013 11:13:59 -0500 From: Tom McNemartmcne...@lmhealth.org Subject: RE: [Histonet] Re: Staining on Alcohol Fixed Smears To: 'Beth Cox'bethc...@gmail.com, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu,levi.fr...@gmail.com levi.fr...@gmail.com Message-ID: E9A90E28259D2F4E84308C5E8EA8F7B4016951E94252@lmhs-exchange Content-Type: text/plain; charset=us-ascii Since there is no exposure to formalin, we do not do antigen retrieval on alcohol fixed specimens. We use the Ventana Benchmark XT and copy the protocol to a new number and use the Wet option rather than paraffin. No retrieval needed. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcne...@lmhealth.org www.LMHealth.org -Original Message- From:histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Beth Cox Sent: Saturday, December 14, 2013 6:30 PM To:histonet@lists.utsouthwestern.edu;levi.fr...@gmail.com Subject: [Histonet] Re: Staining on Alcohol Fixed Smears Levi, I've worked up most of the more common antibodies on alcohol fixed smears. There are two things to remember here: 1) your specimens are alcohol fixed, meaning antigen retrieval needs are different, and 2) they are smears, meaning the cells weren't 'cut open' during microtomy, which impeeds some nuclear staining. Those are two different issues to consider. Some rules of thumb: 1) most cell membrane antigens will stain nicely with minimal changes to the protocols you use for FFPE specimens 2) most cytokeratin antigens stain nicely with little change to your FFPE protocols (if you need to change, start by cutting your antigen retrieval time in half) 3) most nuclear antigens require significant changes from FFPE protocols, and some are essentially impossible to stain using standard antibodies. When I return to work on Monday, I will forward you my protocol 'adjustments' for the antibodies you requested. Beth Cox, HTL/SCT(ASCP)QIHC Vagabond Histotech On 12/14/2013 12:00 PM,histonet-requ...@lists.utsouthwestern.edu wrote: Message: 4 Date: Sat, 14 Dec 2013 18:25:08 +0200 From: Levi Friedlevi.fr...@gmail.com Subject: [Histonet] Staining on Alcohol Fixed Smears. To:histonet@lists.utsouthwestern.edu Message-ID: CA+puqxTXL85pEwvR=sqPQA36tMJPm=sx2kv3gyebrbu0he3...@mail.gmail.com Content-Type: text/plain; charset=ISO-8859-1 Hi Everyone. I am looking to stain alcohol fixed smears with a group of antibodies. The antibodies of particular interest are p63, p53 and ki67. Along with these antibodies I am looking for positive nuclear and cytoplastic staining antibodies. If anyone has any experience in staining alcohol fixed slides your information is very appreciated. All the best. -- Sincerely, - Levi Fried ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Staining on Alcohol Fixed Smears
Levi, I've worked up most of the more common antibodies on alcohol fixed smears. There are two things to remember here: 1) your specimens are alcohol fixed, meaning antigen retrieval needs are different, and 2) they are smears, meaning the cells weren't 'cut open' during microtomy, which impeeds some nuclear staining. Those are two different issues to consider. Some rules of thumb: 1) most cell membrane antigens will stain nicely with minimal changes to the protocols you use for FFPE specimens 2) most cytokeratin antigens stain nicely with little change to your FFPE protocols (if you need to change, start by cutting your antigen retrieval time in half) 3) most nuclear antigens require significant changes from FFPE protocols, and some are essentially impossible to stain using standard antibodies. When I return to work on Monday, I will forward you my protocol 'adjustments' for the antibodies you requested. Beth Cox, HTL/SCT(ASCP)QIHC Vagabond Histotech On 12/14/2013 12:00 PM, histonet-requ...@lists.utsouthwestern.edu wrote: Message: 4 Date: Sat, 14 Dec 2013 18:25:08 +0200 From: Levi Friedlevi.fr...@gmail.com Subject: [Histonet] Staining on Alcohol Fixed Smears. To:histonet@lists.utsouthwestern.edu Message-ID: CA+puqxTXL85pEwvR=sqPQA36tMJPm=sx2kv3gyebrbu0he3...@mail.gmail.com Content-Type: text/plain; charset=ISO-8859-1 Hi Everyone. I am looking to stain alcohol fixed smears with a group of antibodies. The antibodies of particular interest are p63, p53 and ki67. Along with these antibodies I am looking for positive nuclear and cytoplastic staining antibodies. If anyone has any experience in staining alcohol fixed slides your information is very appreciated. All the best. -- Sincerely, - Levi Fried ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Cytopathology Quest - alcohols
Regarding alcohols in cytology: Amber, you are right about those alcohols being equivalents for *shrinkage rates* (and since size is one of the criteria for cyto, shrinkage rates are important) but that does NOT indicate they perform the same. Reagent alcohol (but not all other denatured alcohols) has very similar performance and shrinkage to ethyl alcohol and is considered an good acceptable substitute. 80% isopropal alcohol has similar shrinkage but not quite the same performance as ethyl and is considered an adequate substitute for fixation in emergency situations - the same is true for absolute methyl alcohol. Beth Cox, SCT/HTL(ASCP)QIHC -- Message: 1 Date: Mon, 14 Oct 2013 20:52:25 + From: Ian R Bernard ibern...@uab.edu Subject: RE: [Histonet] Cytopathology Quest. To: Fimbres, Amber afimb...@uci.edu, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Message-ID: d4f4c602b10b9f45b4e9271af6380e161821c...@uabexmb1.ad.uab.edu Content-Type: text/plain; charset=us-ascii Thanks for your reference and input IB -Original Message- From: Fimbres, Amber [mailto:afimb...@uci.edu] Sent: Monday, October 14, 2013 11:58 AM To: Ian R Bernard; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Cytopathology Quest. According to Koss' Diagnostic Cytology and its Histopathologic Bases (1979), 100% methanol is equivalent to 95% ethanol which is equivalent to 95% reagent (denatured) alcohol which is equivalent to 80% propanol which is equivalent to 80% isopropanol. So in theory, if you ran out of 95% reagent alcohol and only had isopropanol on hand, as long as it was 80% it would be OK to use. Have I tried this? Honestly...no. Amber Fimbres, CT(ASCP)HTL -Original Message- From: Ian R Bernard [mailto:ibern...@uab.edu] Sent: Sunday, October 13, 2013 6:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cytopathology Quest. Per text books, for cytology specimens, 95% Ethyl Alcohol is the fixative of choice. Are there any studies or references that approve 95% isopropyl or denatured alcohol as a suitable substitute? Ian Bernard ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Are those sites using the Thermo Fisher SlideMate slide printer also using Thermo slides?
We also installed 3 Thermo SlideMates in April 2012. They have had light use since then because Thermo could not get/install a PrintMate cassette printer to finish the project. We have found the best slide to use is the Colorfrost slides from Statlab. However our print quality is VERY inconsistent and still unsatisfactory. Right now, all three are in having their print heads replaced ($1,000 each) after only 5 months of light use. Like IB, we are also waiting for Thermo to fix the problems. We are definitely dissatisfied with the results and unhappy with the Slidemates. Beth Cox, HTL/SCT(ASCP)QIHC Histology Supervisor St Elizabeth Healthcare Edgewood, KY Subject: RE: [Histonet] Are those sites using the Thermo Fisher Slide Mateslide printer also using Thermo slides? To: Elizabeth Chlipalal...@premierlab.com, 'Harrison, Sandra C.' sandra.harris...@va.gov,histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Message-ID: d4f4c602b10b9f45b4e9271af6380e161418d...@uabexmb4.ad.uab.edu Content-Type: text/plain; charset=us-ascii We have also acquired a Thermo Fisher slide mate/micro writer in Mar/Apr of this year. We have yet to have them work and consistently. The printing quality despite the type slides used is inconsistent. Nevertheless, I am giving them opportunity to fix it since we have invested money. IB USAFA, Colorado Springs. -Original Message- From:histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Wednesday, October 10, 2012 2:12 PM To: 'Harrison, Sandra C.';histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Are those sites using the Thermo Fisher Slide Mate slide printer also using Thermo slides? Sandra We had one slide mate that we purchased few years back, we had problems from the beginning with both fisher and other slides, right now the machine is sitting in a cabinet not being used. To me we found it very unreliable with respects to print quality. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From:histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Harrison, Sandra C. Sent: Wednesday, October 10, 2012 1:00 PM To:histonet@lists.utsouthwestern.edu Subject: [Histonet] Are those sites using the Thermo Fisher Slide Mate slide printer also using Thermo slides? I just purchased 6 Slide Mates. We just began using them yesterday. Are any of you using slides that are NOT Fisher slides on your SlideMates? When you respond, could you answer the following questions? a) Is the print job consistently satisfactory? b) How long have had your SlideMate(s)? c) What difficulties did you have, getting it to consistently print well? d) Approximately how many slides per day you are printing on (each)SlideMate? Thanks, Sandy C. Harrison, HTL (ASCP) Histology Supervisor Minneapolis VA 612-467-2449 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Histonet QIHC
Hi Sarah, I've been working on editing a QIHC Study Guide (sorry, it's not ready for the public yet!) and so I've been paying quite a bit of attention to what is on the QIHC exam now-a-days. I've found that most of the QIHC questions are complex analysis and problem solving type questions. It's not enough to know that facts, you must understand how to use the info you have. Since you have been doing hand-staining and troubleshooting, I expect you will have a good handle on that stuff. I recommend that you get the DAKO Immunohistochemical Staining Methods Educational Guide and Educational Guide to Demasking Antigens (both are free from DAKO, just call them!) and review those. The Dako books provide the most concise, complete info you can find. Review the basic immunology stuff for a refresher. Pay attention to all the various systems for IHC. Pay extra attention to antigen retrieval, there are quite a few questions relating to that. Also, understand how to do the calculations for dilutions and titrations. You can expect a couple oddball questions about IHC on cytology preps and frozen sections. Good Luck! Beth Cox, HTL/SCT(ASCP)QIHC ___ Message: 6 Date: Thu, 9 Feb 2012 15:35:29 + From: Sarah Dysartsdys...@mirnarx.com Subject: [Histonet] QIHC To: histonethistonet@lists.utsouthwestern.edu Message-ID: 8a70a9b2ecdd084dacfe6c59fcf86d5001cfc...@bl2prd0710mb363.namprd07.prod.outlook.com Content-Type: text/plain; charset=us-ascii Hey ya'll. I have been doing IHC for 10 years or so now. Do you think The QIHC exam would be much of a challenge if I went and tried to do it without much studying. I have never used automation to do IHC and have always had to troubleshoot my by hand stains myself. If you do think something would be worth going over what would you suggest? Thanks Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Non-gyn cytology prep - amount processed?
Hi Cathy, Your message came through a little garbled, but I think I get the jist of it. Standard procedure for large volume cytology fluid specimens is to mix the specimen well, and then take off a 50 ml aliquot to centrifuge down and make the slides and cell block. If the specimen is not mixed, you are likely to miss the settled malignant cells. It is not prudent (or recommended!) to centrifuge down the entire specimen. If a specimen were extremely sparsely cellular, I might occasionally spin down two 50 ml aliquots. I agree that if a specimen has set for a while and settled naturally, pouring off the supernatent and using the concentrated precipitate for a 50 ml aliquat can be a nice bonus. BUT, there should have been malignant cells in the WELL-MIXED original aliquot. What prep procedures are you using for the slides?? ThinPrep processing can be subject to problems with these kind of specimens because sometimes the filter becomes 'clogged' with fibrin and protein, and the processor thinks it has cells, but not much is there. ThinPrep may not be the best choice for body fluid specimens. A well done cell block should also have shown cells in it. If you are simply scraping loose cells into a lens paper to process and try to embed for a cell block, I'm not surprised if you lost the cells and only recovered the more prominent fibrin. It's always a good idea to use some method to actually hold (block!) the cells together - such as agar, thrombin-prothrombin, or albumin clot. Beth Cox, SCT/HT(ASCP) _ Message: 3 Date: Thu, 11 Dec 2008 10:10:31 -0800 From: cathy.crump...@tuality.org Subject: [Histonet] Non-gyn cytology prep - amount processed? To: histonet@lists.utsouthwestern.edu Message-ID: of6ce74de0.44dc7092-on8825751c.0063d701-8825751c.0063d...@tuality.org Content-Type: text/plain; charset=ISO-8859-1 Hi all, for those of you who also work with cytology - what is procedure for fluids that have a large volume? How much of thefluid do you process? I am wanting to compare our polic instance this week where we received processed 100 mL of that after making sure it first cell blocks only contained fibrin, but the malignant cells. When we went back to the contai settled after 24 hours so we poured off the top part of t and only processed the gunk at the bottom. The second time t were malignant cells in the cell block. My pathologist is pushin for us to process 100% of the fluids, 100% of the time, saying what we cu get those ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
