[Histonet] immunohisto c-MYC and APC
Morning to All Please need some help. Anyone used the following antibodies on FFPE human tissue. Novocastra mouse monoclonal c-MYC(oncoprotein) clone 9E11 and APC (adenomatous polyposis Coli)clone EMM43. Have tried with and without retrieval. Tried different retrievals No staining. Thanks Marilyn ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Lab Telephone
When our new lab was built they gave us phones that could only be used for dialing out...and none in the smaller, procedure rooms. We argued that the tissue processors and other instruments were often in these procedure rooms and it would be handy if we needed a service tech to walk us through a problem. We ended up with the phones in the procedure rooms and also a phone that allowed incoming calls in the main lab. That way we could call the lab looking for individuals without having to run around looking for them. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartl...@cdc.hhs.gov -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara Sent: Tuesday, August 18, 2009 3:48 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Lab Telephone I'm having a challenge wrapping my head around this one: my boss wants to know why I need a telephone in my new lab in the new building we're moving into in January. Part of this is cost (what isn't a cost issue these days??) but I am to come up with justification for having a telephone IN the lab. The new phones will be cordless but he wants the phone to be in the hallway for what he deems a safety issue (I am not a BSL3 lab). The new phone system will be individual lines for specific locations within our facility. I'm the only tech and I'll have a 22x22' lab and the adjoining (with separate entrance) storage room/volatile storage and I will be doing specimen cut-in in another area altogether. I know I've left out some salient points so if you need more info AND if you have some logical input, please ANSWER offline to me directly. I can't figure out why I would NOT need a telephone... egad! I was asked how many calls (in/out) I make/receive per month as a basis for the decision. OMG. Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Da Phone Issue
Thank you to everyone that replied to my (ridiculous) phone justification message! I have many very good, logical, sensible, common-sense reasons there should be a phone IN the lab and I will use every one of them in a short, bullet-point message to my boss. Another reason Histonet is such a priceless device - albeit usually used for more worthy reasons than justifying having a telephone in a histo lab! Let me see... how long until I can retire? WOA - 18 months! Anybody need a p.r.n. in New England??? Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Da Phone Issue
Is Ohio close enough? Linda Blazek HT (ASCP) Manager/Supervisor GI Pathology of Dayton 7415 Brandt Pike Huber Heights, OH 45424 Phone: (937) 293-4424 ext 7118 Email: lbla...@digestivespecialists.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara Sent: Wednesday, August 19, 2009 8:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Da Phone Issue Thank you to everyone that replied to my (ridiculous) phone justification message! I have many very good, logical, sensible, common-sense reasons there should be a phone IN the lab and I will use every one of them in a short, bullet-point message to my boss. Another reason Histonet is such a priceless device - albeit usually used for more worthy reasons than justifying having a telephone in a histo lab! Let me see... how long until I can retire? WOA - 18 months! Anybody need a p.r.n. in New England??? Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Tech vs Professional
Does anyone out in histology have any documentation from CMS, ASCP, or NSH that breaks down the job duties or areas for what is covered under Tech and professional part of the CPT billing codes. For example is Grossing part of the a profee or tech component. Rick Garnhart HT(ASCP) Memorial Health System Histology Supervisor 1400 E. Boulder St. Colorado Springs, CO 80909 Cell: 719-365-8357 Ph: 719-365-6926 Fax: 719-365-6373 rick.garnh...@memorialhealthsystem.com Mission: To provide the highest quality health care Vision: To create an outstanding health system where patients heal and people thrive Values: Compassion - Integrity - Quality - Respect - Teamwork www.memorialhealthsystem.com The information contained in or attached to this electronic message is privileged and confidential, intended only for the use of the individual(s) named above. If the reader of this message is not the intended recipient, or the employee or agent responsible to deliver it to the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited. If you have received this communication in error, please inform the sender immediately and remove any record of this message.___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Evan's Blue - blood brain barrier
We will be infusing Evan's Blue to assess blood brain barrier disruption in some rats brains. The current plan is to follow this with saline perfusion to clear the vasculature and then perfuse with fixative. Brains will then be sunk in sucrose and frozen for sectioning, but I am wondering if anyone knows if Evan's Blue withstands processing to paraffin? Thanks for any tips/ info, etc. Brett Notice: This e-mail message, together with any attachments, contains information of Merck Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp Dohme or MSD and in Japan, as Banyu - direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] in situ with paraffin sections
Hello I've been reading a few protocols for RNA in situ hybridization with paraffin sections. One of the suggestions is to dehydrate the sections on the first day after rehydration and pre-hybridization processing (para fix, proteinase K, acetylation, pbs washes between all). This seems counter intuitive to me, or at least unnecessary. Any suggestions why this would be done? Our protocol does not involve this, by the way, and it works fine except for the tissue tearing. Emily One of the defining characteristics of modern surgery was that patients ought to survive it. --Peter Stanley, For Fear of Pain: British Surgery, 1790-1850 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] job opening, Minneapolis VA
DEPT. OF VETERAN AFFAIRS MEDICAL CENTER, MINNEPOLIS, MN. Full time Histotech. opportunity at Minneapolis VA. BS or BA in Biology. HT cert. required, HTL preferred. Prefer 5 yr. exp. IHC experience a plus. Effective interpersonal skills required. Holiday, evenings and weekends off. Excellent bene's. Detail oriented. Responsible for technical and procedural operations of the dept., performing quality control, quality improvement and regulatory compliance tasks. Please e-mail resume to Sandra Harrison, Histology Supervisor, at sandra.harris...@va.gov. Principals only. No recruiters please. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] destaining bone
Hello All, I hope someone can assist us with a protocol for destaining some mouse ulna. A student stained his bones for 12 hours in 1% basic fuchsin in graded ethyl alcohol from 80 % , 90 % and then 100% as a prestain then into 100 % ethyl alcohol to destain some prior to pmma plastic embedding and sectioning to look at microdamage in the cortical bone. The bones are over stained and if possible we would like to destain the bone and continue on without getting more specimens. Any suggestions are welcome. Thank you in advance. John Baker and Mathieu Davis John A. Baker The University of Michigan Orthopaedic Research Laboratories Histology Unit 109 Zina Pitcher Place, 2218 BSRB Ann Arbor, MI 48109-2200 734-936-1635 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Amyloid staining on frozen muscle bx's
Hi, What stain works for amyloid on frozen muscle bx tissue? I did the a Congo Red stain without much success. Thanks Sharon sal...@hsc.mb.ca 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] Please recommend xylene substitutes
I have used Clear Rite 3 for may years. It is not as forgiving as xylene when it comes to water, but if you keep it fresh you will be fine. Marianne Kitchell Novus INternational From: histonet-boun...@lists.utsouthwestern.edu on behalf of jennifer cresor mike hough Sent: Tue 8/18/2009 7:37 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Please recommend xylene substitutes Hello All, I am looking into switching to a different xylene substitute. I currently use Slide Brite and am having problems with water in it. Please recommend your favorite. Thank you for your response. Jennifer Temecula, CA jenc...@ca.rr.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTE The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, please be advised that any reading, review, forwarding, dissemination, distribution or copying, of this communication or any attachment(s) are strictly prohibited. If you have received this e-mail in error, please so notify the sender immediately. Also, please delete it and all attachments from any servers or other hard drives. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] bone sections
Hi Everyone Could any one tell me how they keep bone sections adhered to slides when staining. I'Ve tried polysine slides, pva glue slides, different ways of drying the sections and for different lenghts of time but nothing seems to work for every section. I'm only staining them with HE. Thanks Marie O'Brien (liverpool vet path - histology) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] anti-bromodeoxyuridine staining
I am having a problem with my anti-bromodeoxyuridine (BRDU) stain. I have done this for many years on chicken tissues fixed in Notox. Now I have rat tissues (Notox again) and sometimes I get everything staining (all nuclei) and sometimes I get next to nothing to stain. I have recently started cutting all my sections (paraffin) at 4 um. Instead of 5um which may be a partial effect. I am pretty sure that my problem is in the 2.0 N Hydrochloric acid (HCl) step and/or the trypsin step. It seems that if I use freshly prepared HCl, I get a better response than if I have previously prepared and stored my 2.0 N HCl or buy 2.0 N HCl prediluted. This in turn affects the amount of time required in the subsequent trypsin step. Every time I get a new bottle of HCl I need to retitrate the whole stain. Any help here so I am not redoing this into infinity? I am starting to live the movie Groundhog Day. I have never been able to perform this stain without an additional trypsin step even though I have seen it done this way in the literature. Thank you all in advance. Marianne Kitchell Novus International St. Charles, Mo. CONFIDENTIALITY NOTE The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, please be advised that any reading, review, forwarding, dissemination, distribution or copying, of this communication or any attachment(s) are strictly prohibited. If you have received this e-mail in error, please so notify the sender immediately. Also, please delete it and all attachments from any servers or other hard drives. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] bone sections
Are you working with resin or paraffin sections? Jack Sent from my iPhone On Aug 18, 2009, at 6:50 AM, Williams, Sean s.a.willi...@liverpool.ac.uk wrote: Hi Everyone Could any one tell me how they keep bone sections adhered to slides when staining. I'Ve tried polysine slides, pva glue slides, different ways of drying the sections and for different lenghts of time but nothing seems to work for every section. I'm only staining them with HE. Thanks Marie O'Brien (liverpool vet path - histology) ___ 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] in situ with paraffin sections
Emily, Unfortunately, there are a lot of superfluous steps involved in ISH. It reminds me of the initial IPX protocols. There were many unfathomable steps included that seemed to incite a feeling of magic required to obtain results. Simplifying the technique was one of the main reasons why routine labs were able to utilise it. I hope the same will occur with ISH. IPX and ISH are similar: Block the endogenous enzyme Reverse the formalin cross-linking Apply the probe or antibody Demonstrate the bound probe or antibody. Stringency washes might be required if there is a high possibility of cross-hybridisation. Tissue sections will react differently to Molecular biology nucleic acid containing tubes and gels Granted, for DNA hybridisations you will need to denature the DNA strand but overall that is about it. I would keep it simple and go from there. Many of the added steps were added by researchers because they seemed like the right thing to do not because they were required. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Emily Sours Sent: Thursday, 20 August 2009 2:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] in situ with paraffin sections Hello I've been reading a few protocols for RNA in situ hybridization with paraffin sections. One of the suggestions is to dehydrate the sections on the first day after rehydration and pre-hybridization processing (para fix, proteinase K, acetylation, pbs washes between all). This seems counter intuitive to me, or at least unnecessary. Any suggestions why this would be done? Our protocol does not involve this, by the way, and it works fine except for the tissue tearing. Emily One of the defining characteristics of modern surgery was that patients ought to survive it. --Peter Stanley, For Fear of Pain: British Surgery, 1790-1850 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * 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 are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Amyloid staining on frozen muscle bx's
Sharon, For successful amyloid staining with Congo red, formalin fixation might be required. I have not been able to find any references for this except that formalin is used in the demonstration of amyloid in fat aspirations (reference: Duston et al (1987) Am J Med 82:412-414) Have you tried one of the metachromatic dyes (eg Crystal Violet)? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sharon Allen Sent: Thursday, 20 August 2009 6:57 AM To: histo...@pathology.swmed.edu Subject: [Histonet] Amyloid staining on frozen muscle bx's Hi, What stain works for amyloid on frozen muscle bx tissue? I did the a Congo Red stain without much success. Thanks Sharon sal...@hsc.mb.ca * 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 are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] anti-GFP antibody
Hi all, I am looking for a good anti-GFP antibody. I plan on using it for dual immunofluorescence on mouse bone in paraformaldehyde fixed, paraffin embedded sections, most likely with some antigen retrieval. Here's the main problem: I want to use it with a goat-anti-mouse primary and in a separate assay with a rabbit-anti-mouse antibody, both of which are staining mouse bone. In order to save money and optimization time, I would prefer if it were raised in a species other than goat and rabbit so I could use it for both assays. I also want to avoid mouse antibodies to avoid the mouse-on-mouse issue. Alternatively, I could buy two antibodies (rabbit and goat) and use them in the respective assays. Or is there another way that's non-obvious? Thanks, Adam ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
