[Histonet] Hello everyone, I am new to this forum
Hello everyone, I want to introduce myself to all the histotechs here. I have visited this forum before but I finally subscribed today. My name is Valerie, but please call me Val. I am an histotechnologist living in Puerto Rico. I got interested in the field of histology in 2008. I always loved Biology but I really did not wanted to study medicine, so I started to search for careers that were related to medicine but did not required a doctorate degree. First I wanted to study cytotechnology in my country but I did not got admitted to that college because there were many people who applied to the program but a very small group was chosen. I remember that my dermatologist told me that I should consider the career of histotechnology instead because they are not a lot of people prepared in this field. Since there are no histotech schools in Puerto Rico I decided to study this career in the U.S. I graduated in 2009. I took the HTL ASCP test in January of 2010 and successfully passed after my second try. I finally got a job, which I started this month in a private laboratory in my country, but instead of working with histology, I am preparing cytology slides! At the begining I was told I was going to do histology in their central laboratory, but I was sent to another laboratory in a private hospital to perform frozen sections but to my surpise, they also told me I was going to stain cytology stains of fine needle aspirations. I have been a little overwhelmed because my expertise is not in cytology. I may follow the protocol of the laboratory for staining these slides, but I really don't have the knowledge of how to troubleshoot a problem in cytology. For now I have been doing a good job, but I recently had a problem with the staining, because the person who trained me did not gave me clear instructions about when to change the solutions so, I did not changed the H20 saline frequently. I tough it was once a week, but then after making this mistake they told me it was like 3 times a week because this solution gets dirty very quickly and can affect the quality of the details of the stains, (they told me this too late because I already stained a whole rack of slides). Now I have to find some info on cytology so that I can understand this field better and stay competitive because in my country you have to do everything, unlike in the U.S where histotech do histology, cytotech do cytology and the pathologist assistant only do the grossing, in here there are histotech that have to do all. Now, to the frozen sections. I still have not performed a real frozen section by myself yet. I have just practiced using little pieces of chicken and I have gotten good sections but, I need to practice with real tissue because not all the tissues are the same and some are very difficult to cut. I have more experience cutting sections with the microtome, embedding blocks, doing routine HE stains. I have not done an special stain yet, but I have knowledge of all of them because histotech school and the ASCP prepared me to know them, but I have little experience in frozen sectioning. I have the knowledge but not the practice. How ironic is life, I prepared myself 100% for histology and now I am doing things that I though I would never do. Do you have similar experiences? Could you provide me advice or good resources about cytology and frozen sections? Thanks ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Block patient IDs
The new derm path lab I am associated with built their own computerized specimen tracking system and I am pretty sure that the same multi unique identifiers that are printed on the slides are printed on the blocks. They have leica slide and block printers connected to accessioning and grossing by touch screen computers and I believe they are labeled the same. If the block labeling and slide labeling are integrated why not keep them the same? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Terri Brown Sent: Friday, March 12, 2010 9:00 AM To: Nails, Felton; Anne van Binsbergen; histonet-requ...@lists.utsouthwestern.edu; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Block patient IDs I think you will find that CAP is asking if 2 patient identifiers are used throughout the entire process. We engrave the blocks, slides and a computer generated block count log that uses 2 patient identifiers. Terri H. Brown,, HT (ASCP) Pathology Laboratory Manager Northside Hospital Atlanta terri.br...@northside.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Nails, Felton Sent: Friday, March 12, 2010 10:55 AM To: 'Anne van Binsbergen'; histonet-requ...@lists.utsouthwestern.edu; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Block patient IDs It is my understand that the two patient identifier applies to the completed slide not the block. You can verify this by looking at the CAP Checklist for anatomical pathology -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Anne van Binsbergen Sent: Friday, March 12, 2010 9:42 AM To: histonet-requ...@lists.utsouthwestern.edu; histonet@lists.utsouthwestern.edu Subject: [Histonet] Block patient IDs A question for all you CAP fundis: how many patient Identifiers are needed on each paraffin block We currently just write the unique, LIS generated number on the block face, but have recently been advised that CAP and JCIA require not one, but TWO patient IDS on the block Comments please -- Anne van Binsbergen (Hope) Abu Dhabi UAE ___ 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 CONFIDENTIALITY NOTICE: This electronic mail transmission has been sent by Northside Hospital. It may contain information that is confidential, privileged, proprietary, or otherwise legally exempt from disclosure. If you are not the intended recipient, you are hereby notified that you are not authorized to read, print, retain, copy or disseminate this message, any part of it, or any attachments. If you have received this message in error, please delete this message and any attachments from your system without reading the content and notify the sender immediately of the inadvertent transmission. There is no intent on the part of the sender to waive any privilege. ___ 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: SPAM-LOW: RE: [Histonet] Red Chromogen for MITF antibody
Is the Perma Red for hrp? If you switch to an alk.phos detection system you could use fast red which is much redder than AEC. Dako sells ap/red detection systems, but I prefer to use Vector Red for AP. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of McMahon, Loralee A Sent: Friday, March 12, 2010 6:23 AM To: Gareth Blaeuer Davis; histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: RE: [Histonet] Red Chromogen for MITF antibody We use Diagnostic Biosystems Perma Red with the Dako system. www.dbiosys.com Works very well Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Gareth Blaeuer Davis [gareth.da...@hotmail.com] Sent: Thursday, March 11, 2010 7:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Red Chromogen for MITF antibody Hi, Does anyone do MITF immunostains with a Red Chromogen. We are using a Dako autostainer, and in the past have used AEC chromogen on MITF. Our pathologist doesn't like it, because it doesn't get red enough. We would like something we could use with current protocols on the Dako, but can't find anything. Any suggestions? Gareth Blaeuer Davis, HT, BS. Pathology Associates of St. Thomas Nashville, Tn 37205 615-298-4100 _ Hotmail: Free, trusted and rich email service. http://clk.atdmt.com/GBL/go/201469228/direct/01/ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Her-2 Controls
Morning All, My chief pathologist has asked the following question. For all of those doing Her-2 Neu staining for scoring with the Vias system. How many control slides do you cut in advance and how long are they viable if kept refrigerated? Thanks in advance, Rae Ann Staskiewicz ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histonet
dear Val, first my english is not good. I can understandbut the grammar! My way is similar- after studies I wanted to work in microbiologie. But I´ve got a job in Pathologie. First time I was alone in the laboratorie.I had to manage the routine...cutting,staining,preparation and so on...and that was at the beginning after my studies, I had no experiances! Sometimes I thought OH GOD but I think, this is the best way to learn! I have made my mistakes! -but I´ve learned.I´ve never made the same mistakes two times! Now I do my job about 36 years and I teach students in histotechnologie. Val, I think there is no problem to make a mistake at the beginning! For your beginning I wish you good luck. Best wishes Renate ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC on 3 year fixed tissue?
I have to imagine that you are doing the HIER in the FFPE sections? If you have used autoclave unsuccessfully it seems that the epitopes are so cross-linked that they do not respond to the treatment. I would try a piece of fixed tissue before processing, cut it in a very thin slice (about 2 mm maximum) and heat it in distilled water at 65ºC for 30 minutes. Wah it thoroughly and process it as usual. After that, prepare the sections and HIER them in citrate buffer in a steamer and proceed as usual. By doing this you will have to steps were the cross-linkages area going to be weaken and perhaps you will get some acceptable reaction. René J. --- On Sat, 3/13/10, Kevin Egan kevi...@mail.med.upenn.edu wrote: From: Kevin Egan kevi...@mail.med.upenn.edu Subject: [Histonet] IHC on 3 year fixed tissue? To: histonet@lists.utsouthwestern.edu Date: Saturday, March 13, 2010, 2:15 PM Hello Histonetters, I've been asked to perform anti-GFP IHC on some mouse bones which have been sitting in NBF for 3-4 years. My Abcam rabbit polyclonal gets great results in GFP mouse control tissue, but I can't seem to get any results out of these bones. I've tried citrate buffer HIER using autoclave, hotplate, and 65 degree water batch over night, but my signal is non-existent (or really the noise is so huge you can't distinguish).Do the great minds of histonet have any ideas on how to achieve good staining? Is enzymatic epitope retrieval worth a try? Does anyone have a time machine I can use to stop my predecessors from just sticking the bones in NBF and forgetting about them for years? This listserv has helped me countless times, I am truly appreciative of the knowledge and experience available on Histonet. Thanks, Kevin ___ 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] IHC on 3 year fixed tissue?
Hey there. I have done IHC on mouse placentae that have been fixed in NBF for over 2 years and it took me 5-10 minutes of ProK (10 ug/ml in PBS) followed by 60 minutes of HIER in 10 mM citrate buffer (made sure PBS was at pH 6) in a veggie steamer. I had previously done this IHC in a veggie steamer for only 25 minutes in 10 mM citrate buffer (pH 6) for mouse placentae that had only been fixed for 24 hours in NBF. The antigen was a nuclear receptor (AhR); I have found nuclear antigens to be slightly more difficult to retrieve...at least for mouse placenta! Note that the 2-year-fixed tissue did NOT react positively until the 60-minute mark after HIER, although I tested at 25, 35 and 50 minutes.Only 60 minutes HIER plus enzyme retrieval worked, even though quick-fixed tissue worked simply with 25 minutes HIER. Definitely worth a try, if you're desperate. It took me over a year to get this **#%*#%** tissue working! grin Also, used a 1:100 dilution instead of my normal 1:500 dilution of the antibody, so you might want to titrate the antibody, too. Am I being lucid? Have just come home from a most excellent dinner party! Perhaps not such a good idea to give histo-advice on a Saturday nightbut I deeply sympathized with your plight grin. If you have any questions, please feel free to contact me personally, Kevin, either on gmail or at my SLRI account. Jacqui Jacqui Detmar Work phone: 1-416-646-0223 Work fax: 1-416-862-9696 Cell phone:1-647-273-8735 jacquidet...@gmail.com From: histonet-boun...@lists.utsouthwestern.edu on behalf of Rene J Buesa Sent: Sat 3/13/2010 6:17 PM To: histonet@lists.utsouthwestern.edu; Kevin Egan Subject: Re: [Histonet] IHC on 3 year fixed tissue? I have to imagine that you are doing the HIER in the FFPE sections? If you have used autoclave unsuccessfully it seems that the epitopes are so cross-linked that they do not respond to the treatment. I would try a piece of fixed tissue before processing, cut it in a very thin slice (about 2 mm maximum) and heat it in distilled water at 65ºC for 30 minutes. Wah it thoroughly and process it as usual. After that, prepare the sections and HIER them in citrate buffer in a steamer and proceed as usual. By doing this you will have to steps were the cross-linkages area going to be weaken and perhaps you will get some acceptable reaction. René J. --- On Sat, 3/13/10, Kevin Egan kevi...@mail.med.upenn.edu wrote: From: Kevin Egan kevi...@mail.med.upenn.edu Subject: [Histonet] IHC on 3 year fixed tissue? To: histonet@lists.utsouthwestern.edu Date: Saturday, March 13, 2010, 2:15 PM Hello Histonetters, I've been asked to perform anti-GFP IHC on some mouse bones which have been sitting in NBF for 3-4 years. My Abcam rabbit polyclonal gets great results in GFP mouse control tissue, but I can't seem to get any results out of these bones. I've tried citrate buffer HIER using autoclave, hotplate, and 65 degree water batch over night, but my signal is non-existent (or really the noise is so huge you can't distinguish).Do the great minds of histonet have any ideas on how to achieve good staining? Is enzymatic epitope retrieval worth a try? Does anyone have a time machine I can use to stop my predecessors from just sticking the bones in NBF and forgetting about them for years? This listserv has helped me countless times, I am truly appreciative of the knowledge and experience available on Histonet. Thanks, Kevin ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] in situ hybridization histochemistry question about radioisotope
I have routinely seen that we obtain slightly different results (like 10-20% variance) in using 35S dATP for TdT-catalyzed tailing reactions of oligonucleotides from month to month as we use different batches (made fresh monthly) of the 35S dATP. But on a few occasions over the years we have found dramatically reduced results with a particular lot. This happened again in February, and I confirmed by comparing directly between the Feb. March lots in March that the results with the Feb. lot were half that of the March batch (of course, given the half-life of 35S, they would be expected to be down some, but not by half!). The company was good about believing me and not charging us for the March replacement batch, but they have no idea what could cause this to happen occasionally. They check the specific activity, pH, purity, etc. and run a bio-assay that is a binding assay each month, but were not set up to test the enzymatic reaction. I told them that years ago when this happened it was finally determined that the DTT concentration was unusually high and interfered with the reaction, but they discounted this possibility. I guess I should just count my blessings that things are working again now (mercifully, I was able to cling to optimism, based on past experience, that the isotope was the problem, until I could confirm this myself), but it's very frustrating that the company is not motivated to check the efficacy of the item for this particular application (which I would imagine a lot of customers use it for) each month, or to figure out what causes this to happen on occasion (so as to avoid it!)--we lose a lot of time and other expensive reagents (e.g. the TdT) each time this happens. I also don't know how to interpret it--I know that as the enzyme begins to degrade it just means that the tails are shorter, which I can compensate for with a longer film exposure, but I have no idea what's going on when the isotope causes the problem am afraid to use those batches. Does anyone else using ISH have any ideas about what causes this variability? grateful for any thoughts, Susan No virus found in this outgoing message. Checked by AVG - www.avg.com Version: 8.5.436 / Virus Database: 271.1.1/2745 - Release Date: 03/13/10 21:51:00 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
