[Histonet] setting up for staining
Hello! My lab doesn't really do staining (H and E, Masson Trichrome) too often (maybe once a week), but I'd really like to have a set of dishes with all of the plastic bins and what not. Unfortunately, this costs $500 for just one row. Has anyone tried the newer set-ups like this https://us.vwr.com/store/catalog/product.jsp?product_id=4790248 It's remarkably cheaper, but I wonder if that's because it's not as good. We don't have a lot of money, and convincing my boss to even consider this will probably be difficult. I've been using three glass dishes and pouring reagents in and out of them; it's time to move on to an actual set where we can store the reagents in the dish. While I wish we could buy the cool Tissue Tek version (since everyone else has it), it's not feasible at all considering the cost. (Also, Ann, stop lurking I know you're reading this!!) Emily By bitching and bitching and bitching, they could exhaust the drama of their own horror stories. Grow bored. Only then could they accept a new story for their lives. Move forward. -Chuck Palahniuk, Haunted ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Masson trichrome and H and E
Hello! I am doing Masson trichrome manually (not with a machine) and I just found out the kit is $800!! I was thinking of buying the ingredients separately, but why the hell is it so expensive??? Also, the people I was going to borrow their reagents from said their aniline blue is not very good and I wanted to replace it. I only need about 250ml, what brand do you guys prefer? l know I can google this, but I want to know what you guys like and what works best. This is for mouse kidney paraffin sections, 4 to 5 microns. Another question, I did H and E and there is no eosin staining. I think the reagents are pretty old, so I thought that might be a problem. Also because my lab is cheap, they were reusing the xylenes and EtOH for both rehydrating and rehydrating. I told my boss this is probably not a good idea as the end steps will have stain in them. And I also think this is why it didn't work! The EtOH is also really old so who know is the 100% is actually even close to 100% any more. I'm buying new reagents, but if you guys think anything else would help, let me know. Also, shoutout to Ann, I know you're reading these!! Join the list!! Emily By bitching and bitching and bitching, they could exhaust the drama of their own horror stories. Grow bored. Only then could they accept a new story for their lives. Move forward. -Chuck Palahniuk, Haunted ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Re: Embedding
That is so cool!! I wish I had one of those!! By bitching and bitching and bitching, they could exhaust the drama of their own horror stories. Grow bored. Only then could they accept a new story for their lives. Move forward. -Chuck Palahniuk, Haunted On Wed, Jan 21, 2015 at 1:59 PM, histot...@imagesbyhopper.com histot...@imagesbyhopper.com wrote: We also use the para-trimmer. In my view, it is worth its weight in gold! I can melt 5 blocks at a time, works like a charm. I am one who does not mind the wax on the sides, as I am most confident that there is enough paraffin to support the cassette. Michelle Sent from my iPhone On Jan 21, 2015, at 11:41 AM, Morken, Timothy timothy.mor...@ucsf.edu wrote: I agree. We got one of these a couple years ago and the techs love it. It is a heated block on which you rub the cassette. The paraffin melts away. It is especially good for preserving barcodes (but don't press the printed surface on the heat block too long - you can soften the print and cause some damage, but nothing like can happen with scraping). Tim Morken Supervisor, Histology, Electron Microscopy and Neuromuscular Special Studies UC San Francisco Medical Center San Francisco, CA CONFIDENTIALITY NOTICE: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this email message or its attachments. If you believe you have received this email message in error, please contact the sender by reply email and destroy all copies of the original message. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto: histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Goins, Tresa Sent: Wednesday, January 21, 2015 8:29 AM To: gayle.cal...@bresnan.net; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: Embedding I agree with Gayle. We finally purchased a trimmer from Ted Pella - lowest price by far - and are saving our finger joints. The amount of wax remaining on the cassette also appears to depend on the brand of mold used. Tresa -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto: histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Wednesday, January 21, 2015 9:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Embedding After years of never winning the battle of paraffin on cassette edges after embedding, we purchased a paraffin block trimmer. It saves time and the stress on finger joints compared to scraping cassettes daily. No matter how careful we were during embedding to keep excess paraffin off cassette edges, we were never successful. Several vendors have these and you may be able to find a refurbished one. Gayle M. Callis HTL/HT/MT(ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] anti-roll plate damage?
Hey everyone, don't forget to reply all when answering questions! Is there a way to change the reply function for this list so you can reply to the list instead just the person asking a question? I don't want to miss out on any answers and I think for this question I didn't see any but the thank you. Pitt just changed their email system and their virus filter is a little too stringent, so my not getting replies could be due to that. But I would love to have a standard reply to list option. Can we get that set up? Feel free to naysay me if the mods have reasons for not doing so. Emily On Nov 3, 2014 12:34 PM, Bain,Virginia veb...@mdanderson.org wrote: Greetings, Our lab recently got a new cryostat (a cryostar NX50). It sees light to moderate usage (3 - 20 hours a week) and has been operational for about 3 weeks. It has had a host of problems since we received it (shipped with a busted motor and the sensor in the window did not detect when it was shut which lead to some kind of short and killed the lighting system). I am hesitant to contact the rep again (they¹ve told us they¹re tired of hearing from us) until I¹m certain this is not user error of some sort. For the most part we have been getting nice clean sections. Every so often we start getting lines and tears which continue even when the blade is moved or replaced. Twice now we have resolved this issue by rotating the anti-roll plate to a smooth edge. When we encounter this problem I find that the anti-roll plate is jagged to the touch, which is I think what is causing the tears. I see that some people suggest smoothing the anti-roll plate out on an emory board when it gets rough but this seems very early in the life of the anti-roll plate to me. In my previous lab we used the same anti-roll plate without issue for about 2 years with heavy usage (50-70 hours a week) and so I¹m wondering why this anti-roll plate is encountering so much damage? Thanks for the help! -- Virginia Bain Postdoctoral Fellow Richie Lab M D Anderson - Science Park 512-237-6443 ___ 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] Ebola
If formalin didn't kill CJD, what did you use? Just curious. Emily By bitching and bitching and bitching, they could exhaust the drama of their own horror stories. Grow bored. Only then could they accept a new story for their lives. Move forward. -Chuck Palahniuk, Haunted On Thu, Oct 9, 2014 at 4:34 PM, Patsy Ruegg prueg...@hotmail.com wrote: Well said Pam, it is just assumed that formalin will eliminate the biohaz for Ebola, I doubt if that has been conclusively proven yet, remember we only discovered fairly recently that formalin fixation did not protect us from CJD Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting 40864 E Arkansas Ave Bennett, CO 80102 H 303-644-4538 C 720-281-5406 prueg...@hotmail.com pru...@ihctech.net Date: Thu, 9 Oct 2014 20:26:16 + From: mucra...@comcast.net To: tanyaabb...@catholichealth.net Subject: Re: [Histonet] Ebola CC: histonet@lists.utsouthwestern.edu Take Brett's advise and use that as guidleine. We don't know as much as we should about these viruses. Pam - Original Message - From: Tanya Abbott tanyaabb...@catholichealth.net To: Histonet histonet@lists.utsouthwestern.edu Sent: Thursday, October 9, 2014 2:03:47 PM Subject: [Histonet] Ebola Dare I ask?! Are any Pathology labs discussing what to do with specimens/precautions, etc. regarding a person with a potential Ebola infection? Tanya G. Abbott RT (CSMLS) Manager Technologist, Histology/Cytology St. Joseph Medical Center Reading, PA 19603-0316 ph 610-378-2635 fax 610-898-5871 email: tanyaabb...@catholichealth.net This electronic mail and any attached documents are intended solely for the named addressee(s) and contain confidential information. If you are not an addressee, or responsible for delivering this email to an addressee, you have received this email in error and are notified that reading, copying, or disclosing this email is prohibited. If you received this email in error, immediately reply to the sender and delete the message completely from your computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ 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] Is there a Law for refusal of pathology services.
What is the reasoning for this?? I think the insurance company would want a diagnosis! Does this person enjoy surgery for the fun of it and not actually need it? Or maybe she has another problem she doesn't want her insurance to know about? Although if she has insurance she can't be retroactively declined, can she? How odd. Emily On Aug 18, 2014 5:04 PM, Nicole Tatum nic...@dlcjax.com wrote: Please help, We had a patient today who had a punch bx of what is believed to be a clinical dermatofibroma. The patient stated they did not wish for the specimen to be sent for clinical testing. Our ARNP discussed the need for pathology at length and the patient stated she was a nurse and could sign a waiver denying pathology services. I have googled and gooled trying to find any specific law or statue. I can only find information pertaining to research or donated tissue. Stating a person no longer has rights or ownershipto the tissue once consented and removed, but this case is not for research. Could someone pls share an actual law with me. Seems silly to consent to the surgery but not to the diagnosis. Im not sure what to do at this point. Have them sign a document on our company letterhead stating there denial of services? Hold the tissue hold long? Accession it but only do gross description? Charge anything? Any thoughts or imformation would be greatly appreciated. Nicole Tatum HT BSH ___ 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] HS IHC project
I am so jealous of these kids. I didn't get near a microscope until college, let alone a microtome. I hope they have fun! Emily By bitching and bitching and bitching, they could exhaust the drama of their own horror stories. Grow bored. Only then could they accept a new story for their lives. Move forward. -Chuck Palahniuk, Haunted On Mon, Jul 21, 2014 at 2:19 PM, Jennifer MacDonald jmacdon...@mtsac.edu wrote: T cell and B cell markers work well with human spleen. From: Patsy Ruegg pru...@ihctech.net To: Histonet@Lists. Edu histonet@lists.utsouthwestern.edu Date: 07/21/2014 11:08 AM Subject:[Histonet] HS IHC project Sent by:histonet-boun...@lists.utsouthwestern.edu Friends, I am going to help a local HS class (a Biotech class) do a simple IHC project. This class already processes tissue, embeds, sections and does an HE stain. My husband fixed up an old black AO microtome for them to use and they actually cut sections. They cannot use human tissue there so what they have available is mouse spleen already ffpe. I was thinking of using a rab antibody that works on ffpe ms spleen. Ki67 comes to mind? It has been a while since I was in the lab so I am consulting with you all to make sure a rab poly or rab mono Ki67 will indeed stain a ffpe ms spleen? If you have evidence of this can you tell me which ab was used successfully. If you can think of a better rab ab to use on the ffpe ms spleen for this project let me know. It would help if we could keep it simple for antigen retrieval. One of our kind vendors will be donating IHC reagents for this HS class project. I hope the IHC meeting is going well in Vegas, wish I was there. Cheers, Patsy Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting 40864 E Arkansas Ave Bennett, CO 80102 H 303-644-4538 C 720-281-5406 prueg...@hotmail.com pru...@ihctech.net ___ 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] Histology as art!
Hello Histonetters, I'm really looking forward to going to the brand new Morbid Anatomy Museum in NYC, but imagine my surprise when I found some histology in their online gift shop!! http://morbidanatomy.bigcartel.com/category/gifts Histology is beautiful, but it is odd to look at those images on clothing. Emily By bitching and bitching and bitching, they could exhaust the drama of their own horror stories. Grow bored. Only then could they accept a new story for their lives. Move forward. -Chuck Palahniuk, Haunted ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] ISH preparation
That may be what you have to do, as the first day of ISH needs to be RNase free! When I do them, it is 10 slides at a time and we have dishes and a lab bench set aside for RNase free procedures only. Obviously this is not feasible in all workplaces! I would just think about it logically. Use gloves when you can, always. Can you keep the hybridizing part RNase free? I don't know what machines you use for this--we do process the slides manually in glass dishes and then hybridize in a slide container and oven kept for RNase free purposes. After hybridization, you don't really need to be too careful. Now I'm wondering if there's some sort of magical machine that does ISH! Emily By bitching and bitching and bitching, they could exhaust the drama of their own horror stories. Grow bored. Only then could they accept a new story for their lives. Move forward. -Chuck Palahniuk, Haunted On Mon, Jun 23, 2014 at 2:06 PM, jennifer.john...@genzyme.com wrote: Hi Folks! I was hoping to get a little input about preparing samples for ISH. We currently RNAse-away our tools at the time of collection, toss the tissues into formalin, process normally and then cut them using RNAse away cleaned microtomes/forceps on a bath of DEPC treated water (in a cleaned water bath). How do other labs prepare paraffin embedded samples? Do you use special reagents - formalin? Do you clean the processor and use treated reagents there too? Do you wash the slides in DEPC treated water? I am trying to figure out how far we need to go here. We are a high throughput lab and we deal with all kinds of tissues and I don't want to have to take down a processor and keep it RNAse free if I don't have to! I will run the experiments and test different methods, but I was hoping some of you could share your wisdom and give me a place to start. Thanks, JJ Jennifer Johnson Staff Scientist Genzyme Corp. Department of Pathology 5 Mountain Road Framingham, MA 01701-9322 Phn - 508-271-3610 Fax - 508-872-9080 ___ 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] Should I leave histology world
As someone who has been in research (basically being a histologist), I can say that there are NO jobs out there for you. The market is saturated with PhDs. Do not leave your job for a research position unless they can guarantee your salary for years. This will be very unlikely, as getting a grant is super hard nowadays. I actually have to be ceritifed to work in a clinical lab, but I know that after 15 years in a lab, I definitely have the skills, just not the certification to be in a clinical lab. I am working in the office now, and in the lab one day a week after having an R01 for ten years and being the lab manager in a research lab. I'm going to get certification in case this office/lab thing doesn't work out in a few years. I wish there was more money in science but there isn't. So the main point is, either get some skills, or go a different path. Research is not where it's at right now. Although, I am assuming you're in the US, this might not be the case in other countries. Emily By bitching and bitching and bitching, they could exhaust the drama of their own horror stories. Grow bored. Only then could they accept a new story for their lives. Move forward. -Chuck Palahniuk, Haunted On Wed, Jun 4, 2014 at 7:45 AM, joelle weaver joellewea...@hotmail.com wrote: Yes thanks for the perspective. I have a bias towards my own experience, and this seems to be good advice. I work in a molecular based lab now and they are very unaware of what it typically is like in a clinical histopathology lab. Good to point other environments are out there that are clinical, and also that research in general can be very different than clinical settings. Some people are just more suited to certain environments over the other. Joelle Weaver MAOM, HTL (ASCP) QIHC Date: Wed, 4 Jun 2014 01:32:11 + From: koelli...@comcast.net To: joellewea...@hotmail.com CC: timothy.mor...@ucsfmedctr.org; optimusprimehistot...@hotmail.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Should I leave histology world Alpha Histotech- I'll put in my few words even though I'm not active anymore and possibly from different perspective. But also using a few assumptions and if my assumptions are wrong then the rest of what I say is probably meaningless. Not IDíng your e-mail address but if you've worked 3 jobs nightshift including a large reference lab, do you live near a big city? And if so is it a city close to a college or university. Research histology should not be overlooked. You will find many molecular or other such non-histo labs that actually do some or even a lot of histology by non-histology personnel or lab workers. Sometimes it is OK, sometimes even great. Sometimes, and I witnessed it, it is at an embarrassing histo level. I can walk up or down university hallways and see a genetics lab or some other molecular lab and see a microtome or cryostat in there. Sometimes those PI's will send histo work to a core lab. Sometimes they don't want to pay per block so do it (and staining and IHC and FISH) themselves. Someone with even minimal wide-ranging histo experience might be welcomed. No timed block cutting counts. Learn some immunology, genetics, molecular techniques, comparative medicine, physiology, etc, etc along the way. Many places even pay for college level courses while employed there. Just a thought if you are near that kind of area. Ray in Seattle From: joelle weaver joellewea...@hotmail.com To: Timothy Morken timothy.mor...@ucsfmedctr.org, Alpha Histotech optimusprimehistot...@hotmail.com, histonet@lists.utsouthwestern.edu Sent: Tuesday, June 3, 2014 5:55:09 PM Subject: RE: [Histonet] Should I leave histology world It would be a shame to get discouraged now after all the time you have already put into histology. If you still want to work in histology, I might suggest you try to have a conversation with a manager, supervisor or lead tech and see if they are willing to support you. Tell them you want to spend more time cuting to be able to section with high quality at the rate that works for their productivity standards. If you present it as a win-win proposition, see what resources, people and time they are willing to chip in to help get where they would like you to be. Make some metric or rate to achieve in microtomy your goal for the year, and put it into writing ( good for all goals:). Or if that is too uncomfortable , approach someone individually whose microtomy skills you admire , and see if they are willing to provide some tips and guidance off work time. I also went through a NAACLS program. Still at my first real histology job , the realization that this was the actual training became apparent very quickly. I had moments of exhaustion and feeling overwhelmed, but I now feel I was also fortunate to work initially at a pretty high volume place. It was a great breaking in for embedding and microtomy.
Re: [Histonet] cryosectioning, bubbles between slide and section
Hello, This may be a little late, but we use a mix of half 30% sucrose and half OCT to section. It works really well with 14 to 25 micron sections and no cooling or heating of the slide. Our tissue is very small though--embryonic spinal cord or brain from chick or mouse. I just change the way I pick it up to get rid of bubbles. Have you tried rolling the slide upwards when you pick it up (if that makes sense) as opposed to rolling it from left to right (or the opposite)? Also as our cryostat gets older, it tends to get more static and we cannot get rid of it. This causes the tissue to adhere to the slide faster, which definitely causes bubbles. That may the case, but I hope not!! Emily By bitching and bitching and bitching, they could exhaust the drama of their own horror stories. Grow bored. Only then could they accept a new story for their lives. Move forward. -Chuck Palahniuk, Haunted On Fri, Apr 25, 2014 at 2:58 AM, Christina Kreutzer christina.kreutze...@gmail.com wrote: Hello members, I would need some help regarding cryosectioning of spinal cord. I am currently establishing cryosectioning on the cryostat (Leica CM1950) for this tissue and am having some problems. I am cutting 4% PFA fixed and succrose infiltrated spinal cords, embedded in Tissue Tek at 10µm and a temperature of approximately -16°C . I get more or less smoth slices but once I let them adhere to the slide -directly from the knife, after straightening it carefully with a brush - I get massive air bubbles between the slide and the slice. I have experience cutting on cryostats and with different tissues and never have had this problem before. I tried to change the temperature of the chamber and/or chunk and tried to warm the slide before adhering the slice and I tried to cool the slide. But it didn't help. Do you think changing from 30% succrose to a mixture of Tissue Tek/Succrose or even Tissue Tek/PBS would help? Does anybody have an advice? Regards Christina ___ 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
