[Histonet] Macrophage marker in sheep for IHC
Hi there, I am very new to basic sciences research and am planning an experiment to quantify and compare adipose tissue macrophage infiltration in sheep model using immunohistochemistry. I am specifically interested in CD11c as a marker of M1 activation of Macrophages. I have been advised that it is not easy and previous effort by my predecessors have not been very successful in this! From the general google, histonet/histosearch and pubmed search. I have made a list of potential candidates: 1.Clones PG M1 and Clone KP1 (Source : Histonet : http://www.histosearch.com/histonet/Nov03A/RE.HistonetSeepmacrophage.html) 2.EMB11, a mouse anti-human CD68 mAb (Dako, Carpenteria, CA) (source: published paper: http://www.ncbi.nlm.nih.gov/pubmed/16460804 ) 1. Another CD68 antibody Source: http://www.ingentaconnect.com/content/ocean/ajra/2011/0025/0002/art00024) 1. Iba1 (ionized calcium binding adaptor molecule 1) is a 17-kDa EF hand protein that is specifically expressed in macrophages/microglia and is upregulated during the activation of these cells.[Wikipedia] (source: I do not remember where I found this one from but Abcam has several antibodies none of which are predicted for sheep) I am writing to ask for suggestions about which one to go for and any other tips about searching for antibodies or performing IHC in paraffin fixed adipose tissue/liver. Any ideas will be gratefully appreciated as I am very new and isolated in this aspect. Best wishes Vivek Dr Vivek Saroha Clinical lecturer in Child health University of Nottingham This message and any attachment are intended solely for the addressee and may contain confidential information. If you have received this message in error, please send it back to me, and immediately delete it. Please do not use, copy or disclose the information contained in this message or in any attachment. Any views or opinions expressed by the author of this email do not necessarily reflect the views of the University of Nottingham. This message has been checked for viruses but the contents of an attachment may still contain software viruses which could damage your computer system: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation.___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Frozen tissue question
From the research point of view, I've heard of people not fixing tissue before they section it. Since I work with embryonic tissue (which is mostly water!), we always fix our tissue. It definitely is better to not fix tissue when you want to stain with antibodies because anything you do to the sample might affect the antigens. However, since it's impossible to section unfixed tissue, you have to fix it somehow. Our lab once tried to section snap frozen unfixed chick embryo because another lab (our Sauron, if you will) had said that's the best way to get antibody staining. We, however, could not get the tissue to section properly. How that other lab did it is beyond me. Either way, there's never a clear yes or no with protocols, it'll always change due to what you're sectioning. Emily A great book should leave you with many experiences, and slightly exhausted. You should live several lives while reading it. -William Styron On Thu, Jun 23, 2011 at 1:47 PM, Nicole Tatum nic...@dlcjax.com wrote: I run a Mohs lab that processes skin by frozen section. There is no need to use any fixative before hand. But, if you are looking for melanoma or melanocytes, freezing can cause artifact and make it difficult to read slides. Limit the amount of nitrogen you use to freeze the specimen. Some places use alcohol as a fixative after the slide is cut. It will for all purposes remove the water continent from the tissue which will preserve tissue. Hope this helps. Nicole Tatum HT ASCP We have a new doctor in our lab who swears that all frozen tissue must be fixed in formalin with a subsequent sucrose treatment before freezing in OCT because not fixing it will cause the structures to be distorted and you can't get good antibody attachment. In my previous experience, we have done this with tissue that came from an animal that was perfused with formalin before the tissue was removed. However, the majority of my previous frozen specimens were flash frozen in OCT and fixed after sectioning. It is also my understanding that fixation in formalin can cause poor antibody detection because of cross-linking caused by the formalin. I would like to hear some other opinions on this please. The particular specimen we will be working with is skin. Thanks in advance, Joel Reichensperger -- Joel Reichensperger Researcher II Southern Illinois University Plastic Surgery Institute jreichensper...@siumed.edu 217-545-7309 (Office) 217-545-1824 (Fax) ___ 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] Tissue Shriveling in Paraffin
I was cutting some bone/joint tissue and noticed that the cartilaginous portion was concave/indented, instead of flush with the rest of the block surface. Even as I continued to cut that portion always seemed a little sunken into the block face and all the sections crumbled. I didn't seal the block after I was done and when I came back the next day the entire tissue sample was shriveled and pulling away from the paraffin. I'm new, but in the few bone sections I've done I've never had this happen! Any ideas? ~Shelly ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] SV-40 control blocks or slides
Hi, I am looking for a lab or a supplier for SV-40 control blocks or slides. If anyone can help us out it would be appreciated. Thanks Sharon Allen Senior Technologist Neuropathology Lab MS435U Health Sciences Centre 825 Sherbrook St. Winnipeg, Manitoba R3A 1R9 Ph# 787-4615 This email and/or any documents in this transmission is intended for the addressee(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. Ce courriel et tout document dans cette transmission est destiné à la personne ou aux personnes à qui il est adressé. Il peut contenir des informations privilégiées ou confidentielles. Toute utilisation, divulgation, distribution, copie, ou diffusion non autorisée est strictement défendue. Si vous n'êtes pas le destinataire de ce message, veuillez en informer l'expéditeur immédiatement et lui remettre l'original. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Macrophage marker in sheep for IHC
CD68 - clone KP1 has not worked on sheep (also not dogs, cows, horses, and chickens) in my experience. I have found it to stain primates well (and rare cells in pigs and cats). Jan Shivers Senior Scientist Histology/IHC/EM Section Head Pathology Teaching Program University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive...@umn.edu (Confidentiality Notice: This message, together with any attachments, is intended only for the use of the individual or entity to which it is addressed and may contain confidential or privileged information. If you think you have received this message in error, please advise the sender and then delete this message and any attachments immediately.) - Original Message - From: Vivek Saroha vivek.sar...@nottingham.ac.uk To: histonet@lists.utsouthwestern.edu Sent: Friday, June 24, 2011 4:05 AM Subject: [Histonet] Macrophage marker in sheep for IHC Hi there, I am very new to basic sciences research and am planning an experiment to quantify and compare adipose tissue macrophage infiltration in sheep model using immunohistochemistry. I am specifically interested in CD11c as a marker of M1 activation of Macrophages. I have been advised that it is not easy and previous effort by my predecessors have not been very successful in this! From the general google, histonet/histosearch and pubmed search. I have made a list of potential candidates: 1.Clones PG M1 and Clone KP1 (Source : Histonet : http://www.histosearch.com/histonet/Nov03A/RE.HistonetSeepmacrophage.html) 2.EMB11, a mouse anti-human CD68 mAb (Dako, Carpenteria, CA) (source: published paper: http://www.ncbi.nlm.nih.gov/pubmed/16460804 ) 1. Another CD68 antibody Source: http://www.ingentaconnect.com/content/ocean/ajra/2011/0025/0002/art00024) 1. Iba1 (ionized calcium binding adaptor molecule 1) is a 17-kDa EF hand protein that is specifically expressed in macrophages/microglia and is upregulated during the activation of these cells.[Wikipedia] (source: I do not remember where I found this one from but Abcam has several antibodies none of which are predicted for sheep) I am writing to ask for suggestions about which one to go for and any other tips about searching for antibodies or performing IHC in paraffin fixed adipose tissue/liver. Any ideas will be gratefully appreciated as I am very new and isolated in this aspect. Best wishes Vivek Dr Vivek Saroha Clinical lecturer in Child health University of Nottingham This message and any attachment are intended solely for the addressee and may contain confidential information. If you have received this message in error, please send it back to me, and immediately delete it. Please do not use, copy or disclose the information contained in this message or in any attachment. Any views or opinions expressed by the author of this email do not necessarily reflect the views of the University of Nottingham. This message has been checked for viruses but the contents of an attachment may still contain software viruses which could damage your computer system: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation.___ 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] Tissue Shriveling in Paraffin
Why not try resin embedding techniques? Less shrinkage than paraffin, more specimen/block stability in cutting, no damaging effects of decalcification and a many times you have a better and more clear morphological representation. I could help you to achieve this if interested, so feel free to contact me as needed (31-281-1975). Jack Jack Ratliff Senior Histologist, Biomimetic Therapeutics, Inc. Chair, Hard Tissue Committee - National Society for Histotechnology On Jun 24, 2011, at 8:27 AM, Michelle Aono aono...@auburn.edu wrote: I was cutting some bone/joint tissue and noticed that the cartilaginous portion was concave/indented, instead of flush with the rest of the block surface. Even as I continued to cut that portion always seemed a little sunken into the block face and all the sections crumbled. I didn't seal the block after I was done and when I came back the next day the entire tissue sample was shriveled and pulling away from the paraffin. I'm new, but in the few bone sections I've done I've never had this happen! Any ideas? ~Shelly ___ 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] Sirius Red Stain
Why not save yourself some time and simply purchase the staining kit from Polysciences or Dorn and Hart Micredge. Jack On Jun 23, 2011, at 3:48 PM, Jesus Hernandez jesus.w@gmail.com wrote: Dear all, Does anyone have a protocol on how to prepare Sirius Red stain with known concentration. We are unsure about which concentration to use. We are trying to stain collagen. The Sirius Red is in powder-form. Thank you. Jesse Hernandez University of Texas - San Antonio Department of Biomedical Engineering ___ 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] Joplin's article
To those who were not able to open to the site, I tried to copy and paste and hopefully you can read it. This is a very interesting story about Joplin's St John's Regional Medical Center and code Gray. If you still have problems. please visit this link. http://m.kansascity.com/kcstar/db_41535/contentdetail.htm?contentguid=0kA2g6z2src=catfull=true#display By ERIC ADLER and LAURA BAUER The Kansas City Star JOPLIN, Mo. | Looking back, they remember the quiet — like a last, deep breath before death. In the nursery of St. John’s Regional Medical Center, newborns napped in bassinets. Ventilators hummed in an intensive-care unit. In the emergency room, nurse Tracy Hernandez checked an older woman for a stroke, one of the few serious cases in the ER all day. In an operating room down a second-floor hallway, orthopedic surgeon James “Dusty” Smith opened an infected hip. One floor above, John Seay, a 60-year-old mechanic from Welch, Okla., visited with his 83-year-old mother. Frail from congestive heart failure, she doubted she’d be getting better and had picked out a pink dress for her burial. Then, in the west, the air began to spin. Condition Gray. The announcement over the hospital speakers warned of a potential tornado. Prepare. No one panicked. Such calls are routine in Joplin, a zinc- and lead-mining town carved from the rock and fields of Tornado Alley. Nurses pulled shades over windows to shield from flying debris. They rolled equipment from the halls, on the off chance patients would have to be moved there. Off chance, because this storm wasn’t expected to hit them. Visitors watched it on television with nonchalance. Radar showed funnel clouds tracking north. In a neighborhood across from St. John’s, Amanda and Bradley German sat in a friend’s home with their sons, Brody, 6, and James, 9, heedless of the weather alert. Small hail fell. The friend tossed hailstones playfully into the house. “We were joking about it,” Amanda German would say. “We hear the storm sirens all the time.” What no one anticipated was the dark monster developing to the west, two miles outside their windows. The sky turned the green of a violent bruise. Execute Condition Gray: Get patients to safety! • • • It’s been four weeks since an EF-5 tornado slammed to the ground in Joplin, its 200-mph winds scouring a three-quarter-mile-wide, six-mile-long band of devastation. When it was over, this city of 50,000 would reel, broken and bloodied. For the watching world, the image of the hollowed shell of St. John’s was ground zero. For 115 years, the hospital system created by the Catholic order of the Sisters of Mercy healed the community’s illnesses and injuries. In seconds, the nine-story symbol of the city’s strength and caring, built and expanded since 1968, would stand gravely injured itself. Yet in many ways, the story of what happened inside the walls of St. John’s might better stand as a microcosm of the horrors, heroism and humanity that played out across Joplin that night. Witness upon witness recounts a stream of “walking wounded,” individuals impaled by wood, glass or metal, limbs missing, flesh torn from their bodies, lurching toward the hospital. Many remain haunted by the carnage they saw. But that night at the hospital, they said, also will be remembered as one of the city’s proudest. There was the surgeon who operated by flashlight as the hospital crashed around him. The ER doc who plunged a chest tube through the ribs of a young man to keep him from dying. Nurses used their bodies to blanket vulnerable patients from wind-hurled debris. A floor tech plucked a flying man from the air. Employees wielded axes to free drugs from locked cabinets. And then there were the strangers. Hundreds rushed in convoys of pickup trucks, descending on the hospital to speed the wounded away. Hospital visitors left the sides of their dead and dying loved ones to carry fragile patients down blackened hallways, guided by the dim light of cell phones. “We did what we had to do,” said John Seay. • • • At 5:41 p.m., the tornado descends, a black, twisting wall on the western horizon. It splinters houses, strips trees, heaves cars. Hailstones crash through glass like sledgehammers. Rain pounds down in a stinging curtain. Minutes away, St. John’s waits — 183 patients in its 367 licensed beds; some 25 patients in the ER; 100 staff on duty; an unknown number of visitors in the patient rooms, halls and waiting areas. In the ER, Angie Abner, 40 — a paramedic who became a nurse only a year ago, and who has missed work the last two days because of food poisoning — has been doing triage. Now, minutes after the call Execute Condition Gray, she is struggling to get patients to safety, into hallways and away from windows. People, having grown too used to these warnings and then seeing storms peter out, refuse to move. “Folks, this is for your own safety,” Abner belts, emphatic. “You have to listen to me!” A man waves
AW: [Histonet] Tissue Shriveling in Paraffin
That sounds like insufficient infiltration with paraffin. Melt your block again and let it sit in paraffin for a few hours or even over night. Most time this helps to get better sections. Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Michelle Aono Gesendet: Freitag, 24. Juni 2011 15:28 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Tissue Shriveling in Paraffin I was cutting some bone/joint tissue and noticed that the cartilaginous portion was concave/indented, instead of flush with the rest of the block surface. Even as I continued to cut that portion always seemed a little sunken into the block face and all the sections crumbled. I didn't seal the block after I was done and when I came back the next day the entire tissue sample was shriveled and pulling away from the paraffin. I'm new, but in the few bone sections I've done I've never had this happen! Any ideas? ~Shelly ___ 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] Looking for 2-3 week temp Histology assignment in July
Hi Histonetters! I am looking for a temp histology assignment for a couple of weeks in July. I am HT (ascp) and have 17 years experience. Please respond to this email if you are looking for some help, thank you, Jill Jill Cox, HT ASCP ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Invitation to connect on LinkedIn
LinkedIn Adekunle Oluwatosin Adeluwoye requested to add you as a connection on LinkedIn: -- David, I'd like to add you to my professional network on LinkedIn. - Adekunle Oluwatosin Accept invitation from Adekunle Oluwatosin Adeluwoye http://www.linkedin.com/e/yvpgd1-gpbbkjr1-3/qXtGZ0-QiF70UPNqEunZRx9zbUTaXy-_ifnGa0-b4uheRh4MMF/blk/I123429292_13/1BpC5vrmRLoRZcjkkZt5YCpnlOt3RApnhMpmdzgmhxrSNBszYPclYOej8VczgPcz59bRhktn1vsk5CbPkUc3cPe3wOcPcLrCBxbOYWrSlI/EML_comm_afe/ View invitation from Adekunle Oluwatosin Adeluwoye http://www.linkedin.com/e/yvpgd1-gpbbkjr1-3/qXtGZ0-QiF70UPNqEunZRx9zbUTaXy-_ifnGa0-b4uheRh4MMF/blk/I123429292_13/3cNnP8VczAOd3cOckALqnpPbOYWrSlI/svi/ -- DID YOU KNOW LinkedIn can help you find the right service providers using recommendations from your trusted network? Using LinkedIn Services, you can take the risky guesswork out of selecting service providers by reading the recommendations of credible, trustworthy members of your network. http://www.linkedin.com/e/yvpgd1-gpbbkjr1-3/svp/inv-25/ -- (c) 2011, LinkedIn Corporation ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Invitation to connect on LinkedIn
LinkedIn Adekunle Oluwatosin Adeluwoye requested to add you as a connection on LinkedIn: -- David, I'd like to add you to my professional network on LinkedIn. - Adekunle Oluwatosin Accept invitation from Adekunle Oluwatosin Adeluwoye http://www.linkedin.com/e/yvpgd1-gpbbtrfa-4r/qXtGZ0-QiF70UPNqEunZRx9zbUTaXy-_ifnGa0-b4uheRh4MMF/blk/I123431533_13/1BpC5vrmRLoRZcjkkZt5YCpnlOt3RApnhMpmdzgmhxrSNBszYPclYPcPkNcPgPcz59bRhktn1vsk5CbPkUc3cPe3wOcPcLrCBxbOYWrSlI/EML_comm_afe/ View invitation from Adekunle Oluwatosin Adeluwoye http://www.linkedin.com/e/yvpgd1-gpbbtrfa-4r/qXtGZ0-QiF70UPNqEunZRx9zbUTaXy-_ifnGa0-b4uheRh4MMF/blk/I123431533_13/3cNnPcPdj4Pd3cOckALqnpPbOYWrSlI/svi/ -- DID YOU KNOW LinkedIn can help you find the right service providers using recommendations from your trusted network? Using LinkedIn Services, you can take the risky guesswork out of selecting service providers by reading the recommendations of credible, trustworthy members of your network. http://www.linkedin.com/e/yvpgd1-gpbbtrfa-4r/svp/inv-25/ -- (c) 2011, LinkedIn Corporation ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Formula R paraffin
If sometimes you are experiencing compression problems it could be because your paraffin has some xylene contamination, or your blocks are not cold enough when cutting, or you are using a paraffin of lower melting point as needed for the tissue you regularly process. Paraplast X-tra always worked fine for me. Check with them for a harder paraffin instead of changing to a different paraffin altogether. René J. --- On Fri, 6/24/11, Clare Thornton cthorn...@dahlchase.com wrote: From: Clare Thornton cthorn...@dahlchase.com Subject: [Histonet] Formula R paraffin To: 'Histonet@lists.utsouthwestern.edu' Histonet@lists.utsouthwestern.edu Date: Friday, June 24, 2011, 12:08 PM Does anyone have any experience with Formula R paraffin, from Leica, good or bad? We currently use Paraplast X-tra but occasionally have troubles with compression, and we heard Formula R is harder and less likely to compress. Thanks! Clare J. Thornton, HTL(ASCP), QIHC Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthorn...@dahlchase.com ___ 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] HistoGel
Hello, Does anyone out there have any experience with HistoGel? It's Richard Allan/Thermo Fisher. They claim that you can embed scant tissues in the gel and then process, embed, and cut as usual. Just wondering how it works in the real world Michael J. Dessoye, M.S. | Histology Supervisor | Wyoming Valley Health Care System | mjdess...@wvhcs.org mailto:mjdess...@wvhcs.org | 575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1485 | Fax: 570-552-1526 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the originator of the message. This footer also confirms that this email message has been scanned for the presence of computer viruses. Any views expressed in this message are those of the individual sender, except where the sender specifies and with authority, states them to be the views of Wyoming Valley Health Care System. Scanning of this message and addition of this footer is performed by Websense Email Security software in conjunction with virus detection software. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] (no subject)
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[Histonet] RE: HistoGel
We use histogel a lot in our lab. It's a research lab and we use it for a few purposes - pelleting cultured cells then creating multi-culture TMAs for testing antibodies and also pelleting cells from ascites and pleural effusions. Has also been used to process really small samples that could have been lost in the processor through the cassettes. Works quite well. The researchers just put the samples in histogel and give it to me in formalin then I process it as I would regular tissue. Cuts very well too. Katy Message: 3 Date: Fri, 24 Jun 2011 12:26:46 -0400 From: Dessoye, Michael J mjdess...@wvhcs.org Subject: [Histonet] HistoGel To: histonet@lists.utsouthwestern.edu Message-ID: e2547e1cd0ee324488a2940994571efa0401f...@wvhcs-exchange.wvhcs.com Content-Type: text/plain; charset=iso-8859-1 Hello, Does anyone out there have any experience with HistoGel? It's Richard Allan/Thermo Fisher. They claim that you can embed scant tissues in the gel and then process, embed, and cut as usual. Just wondering how it works in the real world Michael J. Dessoye, M.S. | Histology Supervisor | Wyoming Valley Health Care System | mjdess...@wvhcs.org mailto:mjdess...@wvhcs.org | 575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1485 | Fax: 570-552-1526 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Histogel Problem
Hi, I've encountered this problem before in my previous lab. To reduce the Histogel from shrinking that badly, avoid putting the tissue at the very edge of the Histogel, space them out nicely in the middle, providing sufficient amount of extra Histogel between each tissue and surrounding them. A little bit of shrinking usually do not interfere with cutting as long as the whole thing is placed flat on the final paraffin block, the rest will stretch out on the water bath. I find sometimes the gel does not behave well in the water but as long as it is not on top of your tissue when placed on your slide, it should interfere much with staining. Rose -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amos Brooks Sent: Friday, June 24, 2011 10:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histogel Problem Hi, I have a problem with some blocks that were prepared with Histogel. I was hoping someone else might have had a similar problem and figured it out. I took a photo of the blocks that were mads and put them in a Picassa album here: https://picasaweb.google.com/lh/photo/APO3HsIMa2_jPOEs3QRGUjhz3qi22FNPb2i5JJnBCAk?feat=directlink The long short of it is that the blocs were prepared by the researcher for me to process. They are mouse kidneys. Now it is entirely possible for him to have goofed something up in preparing the Histogel blocks. I wasn't there when he did it, but when I looked at them before processing, they all looked fine. (Like the adjacent good one in the photo). When they were processed they were placed in the VIP right next to each other. When I went to embed them this morning all but one of the four looked fine. The one that didn't come out well looked like the Histogel had shrunk up and shriveled around the kidneys. I am sure this will be aweful to cut, and the researcher is going to have a bird over it since this happened with another project previously. I would like to have a decent explanation for him, so if anyone knows what might have happened and has suggestions I would love to hear it (yes vendors too are welcome to answer this of course!). By the way, this was processed on a rather short cycle of 15-20 min per station of graded ETOH from 70% to 100% with 3 xylene stations and 4 paraffin stations (45 min for these). It seems fine for everything else that was on the processor. Just that one Histogel block was the issue. Thank you, Amos ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet /prePFONT face=Verdana color=blue size=1SPAN style=FONT-SIZE: 8pt; COLOR: blue This e-mail, including any attachments, is meant only for the intended recipient and may be a confidential communication or a communication privileged by law. If you received this e-mail in error, any review, use, dissemination, distribution, or copying of this e-mail is strictly prohibited. Please notify the sender immediately of the error by return e-mail and please delete this message from your system. Thank you in advance for your cooperation. /SPAN/FONT/P ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] bluing
Looking to change my bluing step in the HE process to obtain a bluer (less purple) hue to the nuclear detail. What is everyone using in their bluing step?? Thanks for all of your ideas!! This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] bluing
There are several bluing solutions in the market, or you could use lithium carbonate at different concentrations until you find one of your liking. René J. --- On Fri, 6/24/11, Webb, Dorothy L dorothy.l.w...@healthpartners.com wrote: From: Webb, Dorothy L dorothy.l.w...@healthpartners.com Subject: [Histonet] bluing To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Date: Friday, June 24, 2011, 3:20 PM Looking to change my bluing step in the HE process to obtain a bluer (less purple) hue to the nuclear detail. What is everyone using in their bluing step?? Thanks for all of your ideas!! This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] bluing
Hi Dorothy, Try Richard Allan Bluing Reagent. Here's what they say about their product: It is a buffered product that ensures the proper alkalinity (pH=8.0). Unlike ammonia and lithium carbonate, RA's Bluing Reagent does not allow for the pH shift which can affect the crispness of nuclear detail. When I first began supervising this lab 5 years ago, they made from scratch their own hematoxylin and eosin (not to mention buffered formalin.) Unfortunately, the quality of the stain was very spotty and caused the Pathologists a lot of problems. I switched us to the Richard Allan 7211 Hematoxylin, which has beautiful, crisp nuclear detail. We also went with the recommended Richard Allan Clarifier, Bluing and Eosin, as well, so that we could produce a consistently high quality HE every time. In 5 years, we've had very few complaints about the stain from the 8-10 Pathologists we've worked with, except for one occasion when there was some isolated nuclear hazing. We did some detective work and determined that the cause was due to a rack or two that had been placed in the oven without properly removing the excess water or draining of the slides before placing them in the oven. Have a great week-end everybody. It's practically the 1st sunny day we've had, here in Minneapolis, for the past 2 weeks and Saturday and Sunday's forecast looks good, too! Sandy Harrison VA-Minneapolis Supervisor, Anatomical and Surgical Pathology 612-467-2449 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Webb, Dorothy L Sent: Friday, June 24, 2011 2:21 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] bluing Looking to change my bluing step in the HE process to obtain a bluer (less purple) hue to the nuclear detail. What is everyone using in their bluing step?? Thanks for all of your ideas!! This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] blades
Trying to clean up some things hanging out there in our lab and wondering what everyone does with a blade that has been used minimally and tech done for the day with the microtome. Where do you store that blade for use tomorrow or do you toss and not worry about the cost involved? I do not like them sitting on top of the microtome. Any good ideas?? Thanks, as always! This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] blades
Dorothy, I put ours in a 15 mL centrifuge tube with a cap sit it on the base of the microtome for the next use, that way, no one gets cut the blade is able to be used to the fullest of it's potential. :-) Best regards, ~Sean McBride Scientific Specialist Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-8275 (fax) smcbr...@andrew.cmu.edu On Jun 24, 2011, at 4:53 PM, Webb, Dorothy L wrote: Trying to clean up some things hanging out there in our lab and wondering what everyone does with a blade that has been used minimally and tech done for the day with the microtome. Where do you store that blade for use tomorrow or do you toss and not worry about the cost involved? I do not like them sitting on top of the microtome. Any good ideas?? Thanks, as always! This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] blades
I use the cardboards that come in a box of slides. A small piece of tape on the open side and mark it used. I just always make sure I have the blade edge facing the folded part. I know some who will tape this folded board it to the side of their microtome and use it as a trimming blade holder. Vikki On Fri, Jun 24, 2011 at 5:10 PM, Esther C Peters epete...@gmu.edu wrote: We put ours in a small slide box (plastic or styrofoam, 5-25 slides) that is clearly marked Microtome Blades for Facing Blocks to be used another day. Esther C. Peters, Ph.D. Assistant Professor Department of Environmental Science Policy Biology Program/Medical Technology Coordinator George Mason University 4400 University Drive, MSN 5F2 Fairfax, VA 22030- Office: David King Hall 3057 Phone: 703-993-3462 Fax: 703-993-1066 epete...@gmu.edu - Original Message - From: Webb, Dorothy L dorothy.l.w...@healthpartners.com Date: Friday, June 24, 2011 4:53 pm Subject: [Histonet] blades Trying to clean up some things hanging out there in our lab and wondering what everyone does with a blade that has been used minimally and tech done for the day with the microtome. Where do you store that blade for use tomorrow or do you toss and not worry about the cost involved? I do not like them sitting on top of the microtome. Any good ideas?? Thanks, as always! This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967- 6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ 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] blades
We use plastic 5 slide mailers Sent from my iPhone On Jun 24, 2011, at 1:53 PM, Webb, Dorothy L dorothy.l.w...@healthpartners.com wrote: Trying to clean up some things hanging out there in our lab and wondering what everyone does with a blade that has been used minimally and tech done for the day with the microtome. Where do you store that blade for use tomorrow or do you toss and not worry about the cost involved? I do not like them sitting on top of the microtome. Any good ideas?? Thanks, as always! This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HT Position - Irvine, CA
Hi Histonet, We are currently looking for a full-time Histotechnician (preferably HT certified) with experience in IHC. Please read the description below and if you're interested please submit your resume via e-mail to: * eric.velazq...@agendia.com*. *ESSENTIAL DUTIES AND RESPONSIBILITIES* - Performs histology aspects of the lab, which includes cutting tissue(frozen, FFPE), manually staining sections for HE imaging, IHC staining, loading slides on ScanScope for imaging and scoring by pathologist - Maintains laboratory logs so that current information is complete and readily available to staff, including clinical log sheet, ScanScope identification - Performs all aspects of lab support to ensure timely completion of samples. Reports any discrepancies to the Director Of Laboratory Operations. - Performs various maintenance and/or housekeeping tasks to keep laboratory clean, and ensure ease of use. - Cleans and fills water baths, bulk reagents, dump waste containers - Reports any malfunctions of equipment to the Director Of Laboratory Operations. - Washes all laboratory dishes to ensure a clean supply when needed. - Performs data entry tasks in the laboratory and office when needed. - Follows all Agendia, Inc.’s health and safety policies and procedures - Performs other related duties as required or assigned Thank you for your interest. -Eric Velazquez Agendia Inc. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
