[Histonet] RE: Stupid Rabbit primaries!
Sarah, is it a whole serum antibody? Jackson Laboratories has Ig fragments that can be used to block endogenous Ig sites in the tissue and largely eliminate non-specific staining. A well-purified antibody prep should not give much background, and if it is affinity purified it should be almost as good as a monoclonal. Tim Morken From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of sgoe...@mirnarx.com [sgoe...@mirnarx.com] Sent: Tuesday, March 22, 2011 2:04 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Stupid Rabbit primaries! So I haven't had to deal with rabbit polyclonal primaries in a long time because I remember how much the background sucks with them. Unfortunately the only available antibody is a rabbit polyclonal. Does anyone have any suggestions for how to eliminate the background? I have diluted almost to the point of the antigens not showing! Thanks guys and gals!! Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] B5
Hey all, I was just curious how many of you out there still use B5 as a fixative for bone marrows. Thank Jaclyn Pitts, HT(ASCP) Histotechnician Department of Laboratory Medicine and Pathology Mayo Clinic Rochester, MN E-mail: pitts.jac...@mayo.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] please remove me from list. Thank you.
Please remove me from list. Thank you. Dannie Blake ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Verhoeff/Masson's Stain
See there are always differing experiences (see my earlier post) How are you Bryan, well I hope? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 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 Bryan Llewellyn Sent: Wednesday, 23 March 2011 5:48 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Verhoeff/Masson's Stain Verhoeff's staining solution uses an iron mordasnt, which is an oxidising agent, and iodine, another oxidising agent. It is very easy to overoxidise the hematoxylin, particularly if a stock alcoholic solution is used. I always used to preweigh 1 gram hematoxylin and keep in test tubes, then add the required amount of ethanol at the time I made the working solution. Problems I had had with poor staining then stopped. Have you considered switching to an iron resorcin dye type method? They are progressive, much easier to control and work very consistently. Their drawback is that making the solution is a nuisance and staining often needs overnight. I liked Humberstone's variant (greeny blue black elastic) as the solution is stable for several years and improves over that time with shorter staining times. Others like Miller's (black elatic). There are others as well. Bryan Llewellyn John Shelley wrote: > Hello Histonetters, > > I have been asked to work on a project that will require me to do a > Verhoeff/Masson's stain. I have run some samples through the stain from a > protocol that I found on the archives and it is not working to my > satisfaction. I am not getting the nice elastin fibers to show black like > they should. I have looked at the slides just before going into 2% ferric > chloride and it appears to be dull or limited staining. I am using a kit from > PolyScientific and not sure if that might be my problem or not. My question > is, are there another people who are running this special stain combo and > would you be willing to share the procedure and also who you are buying your > reagents from to accomplish the staining. I appreciate any help with this > request. Thanks!!! > > Kind Regards! > > John J Shelley > > > ___ > 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 * 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] RE: Verhoeff/Masson's Stain
Hi all, My experience with Verhoeff stain indicates that poor, muddy staining of elastic fibres results from using too fresh a solution of alcoholic haematoxylin. The usual methods require a 10% alcoholic Hx that is mixed with the other ingredients. I always let the stock 10% alcoholic Hx stand for at least a week before using. Too fresh Hx is also difficult to differentiate (sometimes less than 1 dip is required in 2% ferric chloride). As for controls, a section of skin is best - the odd artery will stain, but more importantly, the fine elastic fibres in the dermis should be easily seen. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 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 John Shelley Sent: Wednesday, 23 March 2011 5:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Verhoeff/Masson's Stain Hi Shirley, I do make it fresh each time and the reagents are brand new from the company. I am using an appendix as my control and trying to get my hands on some aorta but I really think I should see some elastin fiber staining in the appendix. Kind Regards! John J Shelley -Original Message- From: Shirley A. Powell [mailto:powell...@mercer.edu] Sent: Tuesday, March 22, 2011 2:04 PM To: John Shelley Subject: RE: Verhoeff/Masson's Stain Your problem may be that the elastic stain should be made fresh every time, or you are differentiating in ferric chloride too long. Does the elastic stain get mixed right before use? Also check the expiration date of the kit. Shirley -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of John Shelley Sent: Tuesday, March 22, 2011 1:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Verhoeff/Masson's Stain Hello Histonetters, I have been asked to work on a project that will require me to do a Verhoeff/Masson's stain. I have run some samples through the stain from a protocol that I found on the archives and it is not working to my satisfaction. I am not getting the nice elastin fibers to show black like they should. I have looked at the slides just before going into 2% ferric chloride and it appears to be dull or limited staining. I am using a kit from PolyScientific and not sure if that might be my problem or not. My question is, are there another people who are running this special stain combo and would you be willing to share the procedure and also who you are buying your reagents from to accomplish the staining. I appreciate any help with this request. Thanks!!! Kind Regards! John J Shelley ___ 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 * 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] Stupid Rabbit primaries!
I blocked for all of those... Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 -Original Message- From: Sebree Linda A [mailto:lseb...@uwhealth.org] Sent: Tuesday, March 22, 2011 4:13 PM To: Sarah Goebel; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Stupid Rabbit primaries! Do you know for sure that its the Ab and not endogenous biotin, peroxidase or Phosphatase? You can block for all those. Otherwise there are a number of commercial blocking agents on the market...try Biocare for a start (800)799-9499 or better yet, the company that makes your antibody. Good luck. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of sgoe...@mirnarx.com Sent: Tuesday, March 22, 2011 4:04 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Stupid Rabbit primaries! So I haven't had to deal with rabbit polyclonal primaries in a long time because I remember how much the background sucks with them. Unfortunately the only available antibody is a rabbit polyclonal. Does anyone have any suggestions for how to eliminate the background? I have diluted almost to the point of the antigens not showing! Thanks guys and gals!! Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Stupid Rabbit primaries!
Do you know for sure that its the Ab and not endogenous biotin, peroxidase or Phosphatase? You can block for all those. Otherwise there are a number of commercial blocking agents on the market...try Biocare for a start (800)799-9499 or better yet, the company that makes your antibody. Good luck. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of sgoe...@mirnarx.com Sent: Tuesday, March 22, 2011 4:04 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Stupid Rabbit primaries! So I haven't had to deal with rabbit polyclonal primaries in a long time because I remember how much the background sucks with them. Unfortunately the only available antibody is a rabbit polyclonal. Does anyone have any suggestions for how to eliminate the background? I have diluted almost to the point of the antigens not showing! Thanks guys and gals!! Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Stupid Rabbit primaries!
So I haven't had to deal with rabbit polyclonal primaries in a long time because I remember how much the background sucks with them. Unfortunately the only available antibody is a rabbit polyclonal. Does anyone have any suggestions for how to eliminate the background? I have diluted almost to the point of the antigens not showing! Thanks guys and gals!! Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] HercepTest
We use the PT link, which according to dako is considered a waterbath. 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 Cindy Bulmer [cjbul...@sbcglobal.net] Sent: Tuesday, March 22, 2011 1:12 PM To: Histonet Subject: [Histonet] HercepTest I understand the HercepTest calls for a waterbath for the epitope retrieval process... but, I'm wanting to know what type of equipment, ie: waterbath make and model or if people are using the Dako PT link module. Cindy Cynthia Bulmer HT(ASCP)QIHC IHC Supervisor, CTPL Waco, TX ___ 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
FW: [Histonet] Ventilated Specimen Storage
-Original Message- From: Laurie Colbert Sent: Tuesday, March 22, 2011 12:27 PM To: 'Amy Self' Subject: RE: [Histonet] Ventilated Specimen Storage We have a ventilated closet for storing our old specimens. Our lab is two years old, and this was something that I asked for specifically when they were designing the lab. Laurie Colbert -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amy Self Sent: Tuesday, March 22, 2011 8:35 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Ventilated Specimen Storage Hello Histonetters, How many of you have ventilated storage cabinets for storage of specimens? Thanks in advance for all your help, Amy GHS NOTE: The information contained in this message may be privileged, 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, 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 notify us immediately by replying to this message and deleting it from your computer. Thank you. ___ 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] Re: Ventilated Specimen Storage
Amy Self asks: >>How many of you have ventilated storage cabinets for storage of specimens?<< I'm supposing that by "specimens" you mean surgical specimens stored in formalin in closed containers. I've worked in a lot of pathology services in my life, and I think I've never seen such a thing. Most services keep wet tissue for from two to four weeks. Is some regulatory agency going to require it? Bob Richmond Samurai Pathologist Knoxville TN ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Verhoeff/Masson's Stain
Verhoeff's staining solution uses an iron mordasnt, which is an oxidising agent, and iodine, another oxidising agent. It is very easy to overoxidise the hematoxylin, particularly if a stock alcoholic solution is used. I always used to preweigh 1 gram hematoxylin and keep in test tubes, then add the required amount of ethanol at the time I made the working solution. Problems I had had with poor staining then stopped. Have you considered switching to an iron resorcin dye type method? They are progressive, much easier to control and work very consistently. Their drawback is that making the solution is a nuisance and staining often needs overnight. I liked Humberstone's variant (greeny blue black elastic) as the solution is stable for several years and improves over that time with shorter staining times. Others like Miller's (black elatic). There are others as well. Bryan Llewellyn John Shelley wrote: Hello Histonetters, I have been asked to work on a project that will require me to do a Verhoeff/Masson's stain. I have run some samples through the stain from a protocol that I found on the archives and it is not working to my satisfaction. I am not getting the nice elastin fibers to show black like they should. I have looked at the slides just before going into 2% ferric chloride and it appears to be dull or limited staining. I am using a kit from PolyScientific and not sure if that might be my problem or not. My question is, are there another people who are running this special stain combo and would you be willing to share the procedure and also who you are buying your reagents from to accomplish the staining. I appreciate any help with this request. Thanks!!! Kind Regards! John J Shelley ___ 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] Mycoplasma stain
Hi Histonet- Does anyone know of a good stain for mycoplasma? Our pathologist wanted to try an overnight Giemsa stain, but I wanted to check and see if anyone knew of another stain that might work well. Thanks! Erin Sarricks, HT (ASCP) Histology Laboratory Technician USAMRICD Comparative Pathology Branch Office: Bldg E-3081 Room 178 E-mail: erin.p.sarri...@us.army.mil Phone: 410-436-1967 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] about decalcification
Hello Histonet, Does anyone know procedure of formic acid with sodium citrate decalcification method. ? i need to all of steps of procudure; fixation, decalcification and tissue processing. I will decalcificate rabbit extremites. Thanks in advance. -- Mehmet Fatih BOZKURT, DVM, PhD Afyon Kocatepe University Faculty of Veterinary Medicine Department of Pathology 03030, ANS Campus Afyonkarahisar-TURKEY Tel: +902722281312-109 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] chemicals that are made in the lab
Hi everyone, just was wondering how you determine the expiration dates for the graded alcohols in the lab that you make. Do you take the expiration date off of the reagent alcohol bottles? Also, I use a recycler and recycle Xylene and Alcohol, what expiration dates do you all choose for these? Thanks in advance, Michele Carr ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Verhoeff/Masson's Stain
Hi Shirley, I do make it fresh each time and the reagents are brand new from the company. I am using an appendix as my control and trying to get my hands on some aorta but I really think I should see some elastin fiber staining in the appendix. Kind Regards! John J Shelley -Original Message- From: Shirley A. Powell [mailto:powell...@mercer.edu] Sent: Tuesday, March 22, 2011 2:04 PM To: John Shelley Subject: RE: Verhoeff/Masson's Stain Your problem may be that the elastic stain should be made fresh every time, or you are differentiating in ferric chloride too long. Does the elastic stain get mixed right before use? Also check the expiration date of the kit. Shirley -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of John Shelley Sent: Tuesday, March 22, 2011 1:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Verhoeff/Masson's Stain Hello Histonetters, I have been asked to work on a project that will require me to do a Verhoeff/Masson's stain. I have run some samples through the stain from a protocol that I found on the archives and it is not working to my satisfaction. I am not getting the nice elastin fibers to show black like they should. I have looked at the slides just before going into 2% ferric chloride and it appears to be dull or limited staining. I am using a kit from PolyScientific and not sure if that might be my problem or not. My question is, are there another people who are running this special stain combo and would you be willing to share the procedure and also who you are buying your reagents from to accomplish the staining. I appreciate any help with this request. Thanks!!! Kind Regards! John J Shelley ___ 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] Verhoeff/Masson's Stain
John We do this stain all of the time, but we make up our reagents from scratch we don't use a kit. My suggestion is to make up your elastic stain fresh. We make up our elastic stain fresh and use it only once (or for that day) and then discard. I have attached our procedure in a different e-mail. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of John Shelley Sent: Tuesday, March 22, 2011 11:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Verhoeff/Masson's Stain Hello Histonetters, I have been asked to work on a project that will require me to do a Verhoeff/Masson's stain. I have run some samples through the stain from a protocol that I found on the archives and it is not working to my satisfaction. I am not getting the nice elastin fibers to show black like they should. I have looked at the slides just before going into 2% ferric chloride and it appears to be dull or limited staining. I am using a kit from PolyScientific and not sure if that might be my problem or not. My question is, are there another people who are running this special stain combo and would you be willing to share the procedure and also who you are buying your reagents from to accomplish the staining. I appreciate any help with this request. Thanks!!! Kind Regards! John J Shelley ___ 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] Ventilated Specimen Storage
We do. One in Histology and one in the Morgue. Debbie M. Boyd, HT(ASCP) l Chief Histologist l Southside Regional Medical Center I 200 Medical Park Boulevard l Petersburg, Va. 23805 l T: 804-765-5050 l F: 804-765-5582 l dkb...@chs.net Amy Self Sent by: histonet-boun...@lists.utsouthwestern.edu 03/22/2011 11:37 AM To "'histonet@lists.utsouthwestern.edu'" cc Subject [Histonet] Ventilated Specimen Storage Hello Histonetters, How many of you have ventilated storage cabinets for storage of specimens? Thanks in advance for all your help, Amy GHS NOTE: The information contained in this message may be privileged, 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, 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 notify us immediately by replying to this message and deleting it from your computer. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Disclaimer: This electronic message may contain information that is Proprietary, Confidential, or legally privileged or protected. It is intended only for the use of the individual(s) and entity named in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from your computer. Do not deliver, distribute or copy this message and do not disclose its contents or take any action in reliance on the information it contains. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Verhoeff/Masson's Stain
Hello Histonetters, I have been asked to work on a project that will require me to do a Verhoeff/Masson's stain. I have run some samples through the stain from a protocol that I found on the archives and it is not working to my satisfaction. I am not getting the nice elastin fibers to show black like they should. I have looked at the slides just before going into 2% ferric chloride and it appears to be dull or limited staining. I am using a kit from PolyScientific and not sure if that might be my problem or not. My question is, are there another people who are running this special stain combo and would you be willing to share the procedure and also who you are buying your reagents from to accomplish the staining. I appreciate any help with this request. Thanks!!! Kind Regards! John J Shelley ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HercepTest
Hi Cindy, The protocol is very specific, requiring you use a water bath @ 95 deg for 40 minutes (I think) in the epitope retrieval solution provided. Ashley Troutman BS, HT(ASCP) QIHC Immunohistochemistry Supervisor Vanderbilt University Histopathology 1301 Medical Center Drive TVC 4531 Nashville, TN 37232 (Office) 615-875-3311 (Lab) 615-343-9134 Message: 9 Date: Tue, 22 Mar 2011 09:51:33 -0700 (PDT) From: Cindy Bulmer Subject: [Histonet] HercepTest To: Histonet Message-ID: <614879.51084...@web82307.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello Histoland, ? Anyone using the HercepTest for the Dako autostainer, Code # K5207? If so, what equipment are you using for the Epitope retrieval process? ? Cindy Cynthia Bulmer HT(ASCP)QIHC IHC Supervisor, CTPL Waco, TX ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] vytec formalin neutralizer
Hi Everyone, Is there any one out there who is neutralizing the 10% formalin with Vytec neutralizer? How are you disposing the formalin after neutralization? Need some information regarding this. Thanks in advance Nirmala Holy Name Medical Center is the recipient of: Magnet Recognition for Excellence in Patient Care, American Nurses Credentialing Center 100 Best Places to Work in Healthcare, Ranked Fourth Nationally by Modern Healthcare Best Places to Work in New Jersey, NJBIZ Awards for Emergency, Outpatient and Inpatient Service Excellence, J.D. Power Distinguished Hospital Awards for Clinical Excellence, HealthGrades Excellence Awards for Stroke, Gastrointestinal and Pulmonary Care, HealthGrades Best in Value Award, Data Advantage, LLC Chest Pain Center Accreditation, Society of Chest Pain Centers Primary Stroke Center Designation, The Joint Commission and NJ Department of Health and Human Services Warning: The information contained in this message is privileged and CONFIDENTIAL and is intended only for the use of the addressee above. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or taking of any action in reliance on the content of this message is strictly prohibited. If you have received this communication in error, please notify the sender by replying to this message, and then delete it from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] HercepTest
Cindy I use a Fisher Isotemp 205 waterbath. The end result is beautiful. Cindy Cindy Pyse, CLT, HT (ASCP) Laboratory/Histology Supervisor X-Cell Laboratories 716-250-9235 e-mail cp...@x-celllab.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cindy Bulmer Sent: Tuesday, March 22, 2011 1:12 PM To: Histonet Subject: [Histonet] HercepTest I understand the HercepTest calls for a waterbath for the epitope retrieval process... but, I'm wanting to know what type of equipment, ie: waterbath make and model or if people are using the Dako PT link module. Cindy Cynthia Bulmer HT(ASCP)QIHC IHC Supervisor, CTPL Waco, TX ___ 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] HercepTest
We use the PT Link unit for Hercept Test Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cindy Bulmer Sent: Tuesday, March 22, 2011 11:12 AM To: Histonet Subject: [Histonet] HercepTest I understand the HercepTest calls for a waterbath for the epitope retrieval process... but, I'm wanting to know what type of equipment, ie: waterbath make and model or if people are using the Dako PT link module. Cindy Cynthia Bulmer HT(ASCP)QIHC IHC Supervisor, CTPL Waco, TX ___ 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] HercepTest
I understand the HercepTest calls for a waterbath for the epitope retrieval process... but, I'm wanting to know what type of equipment, ie: waterbath make and model or if people are using the Dako PT link module. Cindy Cynthia Bulmer HT(ASCP)QIHC IHC Supervisor, CTPL Waco, TX ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HercepTest
Hello Histoland, Anyone using the HercepTest for the Dako autostainer, Code # K5207? If so, what equipment are you using for the Epitope retrieval process? Cindy Cynthia Bulmer HT(ASCP)QIHC IHC Supervisor, CTPL Waco, TX ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Grossing Station Recommendation
I can't recommend any specific brand - it's rare to see a grossing station in the small pathology services I work in - but if I were buying one I'd want: 1. First and foremost, adequate ventilation - a louvered vent at hand level that pulls the formaldehyde fumes away from your hands as you work. No overhead kitchen hoods. 2. I think the words "ergonomic" and "pathologist' aren't supposed to be used in the same sentence, but it's awfully nice to have the table rack up and down for pathologists of varying heights. If that isn't possible, at least some sort of elevated table. 3. Most grossing stations are inadequately lighted, at least for my elderly eyes. 4. I don't like to gross with water running over the table and spraying potentially infectious aerosols up into my face. Bob Richmond Samurai Pathologist Knoxville TN ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Ventilated Specimen Storage
Some people just have no sense of humor!! Glad it didn't fall apart with the kick. Reminded me of when the safey office came around asking what "RACE" stood for. He didn't think it was funny at all when I said, "Run around carelessly everywhere" Happy Tuesday, ya'll! Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax Confidentiality Notice: This e-mail, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Ventilated Specimen Storage
It isn't Friday but I'm going on vacation at 2:50PM this afternoon so I'm taking the liberty. This reminds me of a funny CAP story...long time ago and in a galaxy far away (Iowa) we used to store or wet tissue after grossing in plastic garbage cans near a window. Over the years the cans looked a little worse for the wear and they were cracked and had some holes in them. When the CAP inspector asked my labmate is we stored the specimens in a ventilated containers she kicked a can and said, '"why of course we do". He was not amused. Andi On Mar 22, 2011, at 8:35 AM, Amy Self wrote: > Hello Histonetters, > > How many of you have ventilated storage cabinets for storage of specimens? > > Thanks in advance for all your help, > > Amy > GHS > NOTE: > The information contained in this message may be privileged, 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, 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 notify us immediately by > replying to this message and deleting it from your computer. > Thank you. > ___ > 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] RE: muscle bxs
Poly Scientific sells this Walter Benton HT(ASCP)QIHC Histology Supervisor Chesapeake Urology Associates 806 Landmark Drive, Suite 126 (All Deliveries to Suite 127) Glen Burnie, MD 21061 443-471-5850 (Direct) 410-768-5961 (Lab) 410-768-5965 (Fax) wben...@cua.md From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Margiotta, Michele [mmargio...@bmhmc.org] Sent: Tuesday, March 22, 2011 11:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] muscle bxs Hi All, Quick question for those of you that do muscle biopsies. Do you make up the 4% paraformaldehyde or buy it from a vendor. If you make it up, would you share the recipe for it, please? If you purchase it, which vendor do you use? Thanks, Michele BMHMC DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, 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 sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: The information contained in this electronic message is intended solely for the personal and confidential use of the designated recipient(s) named above and may contain information that is protected from disclosure under applicable law. If you are not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this transmission is strictly prohibited. If you have received this transmission in error, please notify the transmitting person/department immediately by email or telephone (410) 581-5881 and delete the message without making a copy. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Recycled Product
Hi. I am wondering if anyone is using recycled product w/ a Leica Bond immuno stainer. And if so, what kind of results are you getting? Any feedback would be appreciated! Thanks in advance Teresa Beal ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Ventilated Specimen Storage
Hello Histonetters, How many of you have ventilated storage cabinets for storage of specimens? Thanks in advance for all your help, Amy GHS NOTE: The information contained in this message may be privileged, 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, 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 notify us immediately by replying to this message and deleting it from your computer. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] muscle bxs
Hi All, Quick question for those of you that do muscle biopsies. Do you make up the 4% paraformaldehyde or buy it from a vendor. If you make it up, would you share the recipe for it, please? If you purchase it, which vendor do you use? Thanks, Michele BMHMC DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, 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 sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Frozen sectioning of pig skin
Saro, I'm certain there are differing thoughts on what to do about your tissue. Was the tissue fixed before it was frozen? How was it stored for the year it was in the freezer? Could be you have a piece of freeze dried pig skin that (if it cuts at all) will look awful. Here is what I would do and I'll probably take some heat (no pun intended) on this but this is what I would do: I'd drop the frozen chunk into fixative and let it thaw and then fix - time depends on the size of the chunk. After fixing I'd rinse the tissue, blot and drop into sucrose to cryoprotect. It is done when it sinks to the bottom of the sucrose. Blot. Then I would re-freeze in a frozen embedding medium like OCT or whatever you have - I like to put the tissue into some OCT and let it sit there for a while before freezing to absorb some of the OCT. I've had difficulty when there is sucrose solution on the outside of the tissue when it is frozen - makes it hard to get a decent section and there is much cursing going on in the lab. Andi Grantham On Mar 22, 2011, at 7:19 AM, Bascaramurty, Saro wrote: > No, I did not. We are using Fisher's Thermo Shandon brand Cryomatrix. We will > order this for next time and I will try and re-do the embedding after thawing > it. Should I lightly rinse the block and pat-dry before re-embedding? > > Thank you, > Saro > > -Original Message- > From: Joel Israel [mailto:jo...@mcclainlab.com] > Sent: March 22, 2011 9:22 AM > To: Bascaramurty, Saro > Subject: RE: [Histonet] Frozen sectioning of pig skin > > Did you thaw the tissue before you embedded in media? If not, try it. > I use NEG-50 from Richard-Allan Scientific to embed frozen tissue. I have > found it to be superior to others. > > -Original Message- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bascaramurty, > Saro > Sent: Tuesday, March 22, 2011 10:00 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Frozen sectioning of pig skin > > > I have sectioned some formalin fixed, paraffin embedded pig skin blocks in > the past, but never the fresh frozen pig skin blocks. The samples were frozen > by the researcher in liquid nitrogen, but not covered in cryomatrix and have > been stored in the deep freezer for over a year. I have now embedded those > blocks in cryomatrix and wanted to prepare sections at 30um and 6-8um > thickness. When I tried it, I had problems getting good quality sections in > spite of using brand new blades. I tried lowering the temperature from -18 > deg.C to -22 / -25 deg.C. I still couldn't get quality sections consistently. > If anyone from histoland has some good tips to improve the quality of these > sections, I would greatly appreciate it. > > > Thank you, > > Saro Bascaramurty > > Technical Officer > Institute for Biodiagnostics > National Research Council > 435 Ellice Avenue, > Winnipeg, Manitoba. R3B 1Y6 > > Tel: 204-984-7166 > Fax:204-984-6978 > email:saro.bascaramu...@nrc-cnrc.gc.ca > > > ___ > 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] how to bleach melanoma cases without bad effect on IHC
We have actually performed a bleach step before staining. The only affect I have seen ( and we have only done a few antibodies) is that the tissue gets pretty chewed up. Especially if there is a pretreatment step prior to the IHC staining. 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 Cynthia Pyse [cp...@x-celllab.com] Sent: Tuesday, March 22, 2011 10:01 AM To: 'Shahram Sabeti'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] how to bleach melanoma cases without bad effect on IHC Sabeti I use the AEC chromogen (Dako catalog #K346989) for my melanoma cases. I use a aqueous mounting media, placed on the tissue only, dry it completely on a hot plate. When dry I coverslip it with permanent mounting media(don't place it in xylene), dry coverslip. Works great, and the color doesn't fade. Cindy Cindy Pyse, CLT, HT (ASCP) Laboratory/Histology Supervisor X-Cell Laboratories 716-250-9235 e-mail cp...@x-celllab.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Shahram Sabeti Sent: Tuesday, March 22, 2011 4:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] how to bleach melanoma cases without bad effect on IHC hello dear fellows, i have an IHC project on paraffin embedded melanoma cases; some of them being heavily pigmented. i have seen many protocols based on H2O2 to bleach the pigmented ones, but i am not sure about any possible effect on antigen retrieval.please guide me . yours, sabeti ___ 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] comprehensive list?
The list is still available online from Novocastra. Here is the link:
http://www.ebiotrade.com/buyf/Novocastra/data/xreact/xreact.pdf
From: histonet-boun...@lists.utsouthwestern.edu
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg
[pru...@ihctech.net]
Sent: Monday, March 21, 2011 11:48 AM
To: 'Rene J Buesa'; 'Histonet'; 'TomTruscott'
Subject: RE: [Histonet] comprehensive list?
Novacastra had a cross reactivity list for their abs which I find very
helpful, I have the old list but can't seem to find anyone who knows about
it recently, I got it years ago when the instrument was still with the AU
Co. Vision BioSystems, I will ask Leica reps to see if they know about this
list.
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net
This email is confidential and intended solely for the use of the Person(s)
('the intended recipient') to whom it was addressed. Any views or opinions
presented are solely those of the author. It may contain information that is
privileged & confidential within the meaning of applicable law. Accordingly
any dissemination, distribution, copying, or other use of this message, or
any of its contents, by any person other than the intended recipient may
constitute a breach of civil or criminal law and is strictly prohibited. If
you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Friday, March 18, 2011 11:30 AM
To: Histonet; TomTruscott
Subject: Re: [Histonet] comprehensive list?
Try your DAKO representative. NOVOCASTRA also used to have such a list for
their Abs.
René J.
--- On Fri, 3/18/11, Truscott, Tom wrote:
From: Truscott, Tom
Subject: [Histonet] comprehensive list?
To: "Histonet"
Date: Friday, March 18, 2011, 12:02 PM
Hi All, Is there a comprehensive list in one location somewhere that gives
information on all or most antibodies on how they work with FFPE or FS or
other fixatives, so that we don't have to try to many approaches when we get
a new antibody to try. Thanks, Tom T
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RE: [Histonet] Frozen sectioning of pig skin
No, I did not. We are using Fisher's Thermo Shandon brand Cryomatrix. We will order this for next time and I will try and re-do the embedding after thawing it. Should I lightly rinse the block and pat-dry before re-embedding? Thank you, Saro -Original Message- From: Joel Israel [mailto:jo...@mcclainlab.com] Sent: March 22, 2011 9:22 AM To: Bascaramurty, Saro Subject: RE: [Histonet] Frozen sectioning of pig skin Did you thaw the tissue before you embedded in media? If not, try it. I use NEG-50 from Richard-Allan Scientific to embed frozen tissue. I have found it to be superior to others. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bascaramurty, Saro Sent: Tuesday, March 22, 2011 10:00 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Frozen sectioning of pig skin I have sectioned some formalin fixed, paraffin embedded pig skin blocks in the past, but never the fresh frozen pig skin blocks. The samples were frozen by the researcher in liquid nitrogen, but not covered in cryomatrix and have been stored in the deep freezer for over a year. I have now embedded those blocks in cryomatrix and wanted to prepare sections at 30um and 6-8um thickness. When I tried it, I had problems getting good quality sections in spite of using brand new blades. I tried lowering the temperature from -18 deg.C to -22 / -25 deg.C. I still couldn't get quality sections consistently. If anyone from histoland has some good tips to improve the quality of these sections, I would greatly appreciate it. Thank you, Saro Bascaramurty Technical Officer Institute for Biodiagnostics National Research Council 435 Ellice Avenue, Winnipeg, Manitoba. R3B 1Y6 Tel: 204-984-7166 Fax:204-984-6978 email:saro.bascaramu...@nrc-cnrc.gc.ca ___ 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] how to bleach melanoma cases without bad effect on IHC
Sabeti I use the AEC chromogen (Dako catalog #K346989) for my melanoma cases. I use a aqueous mounting media, placed on the tissue only, dry it completely on a hot plate. When dry I coverslip it with permanent mounting media(don't place it in xylene), dry coverslip. Works great, and the color doesn't fade. Cindy Cindy Pyse, CLT, HT (ASCP) Laboratory/Histology Supervisor X-Cell Laboratories 716-250-9235 e-mail cp...@x-celllab.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Shahram Sabeti Sent: Tuesday, March 22, 2011 4:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] how to bleach melanoma cases without bad effect on IHC hello dear fellows, i have an IHC project on paraffin embedded melanoma cases; some of them being heavily pigmented. i have seen many protocols based on H2O2 to bleach the pigmented ones, but i am not sure about any possible effect on antigen retrieval.please guide me . yours, sabeti ___ 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] Frozen sectioning of pig skin
I have sectioned some formalin fixed, paraffin embedded pig skin blocks in the past, but never the fresh frozen pig skin blocks. The samples were frozen by the researcher in liquid nitrogen, but not covered in cryomatrix and have been stored in the deep freezer for over a year. I have now embedded those blocks in cryomatrix and wanted to prepare sections at 30um and 6-8um thickness. When I tried it, I had problems getting good quality sections in spite of using brand new blades. I tried lowering the temperature from -18 deg.C to -22 / -25 deg.C. I still couldn't get quality sections consistently. If anyone from histoland has some good tips to improve the quality of these sections, I would greatly appreciate it. Thank you, Saro Bascaramurty Technical Officer Institute for Biodiagnostics National Research Council 435 Ellice Avenue, Winnipeg, Manitoba. R3B 1Y6 Tel: 204-984-7166 Fax:204-984-6978 email:saro.bascaramu...@nrc-cnrc.gc.ca ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Heck
Hi Helen, Can you check the blocks I sent you and see if you have 3 blocks marked EXP 0012. We have so many back up systems in place that it is almost as if I never received these blocks but I have to check anyway. Thanks Jo-Ann Bader Histology Co-Ordinator Goodman Cancer Centre McGill University 1160 Pine Ave. W - Rm 312 (3355) Montreal, QC, Canada H3G 1Y6 Tel: 514-398-8270 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Beckman Coulter HmX-Running the Analyzer without DMS
Our computer went down and we are waiting for service. Is there a way we can run the HmX? In the past before computers all analyzers were able to work just fine and print the results. Kaylee McCaffrey MedWrench Product Manager ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] how to bleach melanoma cases without bad effect on IHC
Sabeti, If you have the option of using a detection system with a chromogen other than DAB, i.e. Alkaline Phosphatase Red, that is by far the easiest way to get around the pigment issue. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Shahram Sabeti Sent: Tuesday, March 22, 2011 3:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] how to bleach melanoma cases without bad effect on IHC hello dear fellows, i have an IHC project on paraffin embedded melanoma cases; some of them being heavily pigmented. i have seen many protocols based on H2O2 to bleach the pigmented ones, but i am not sure about any possible effect on antigen retrieval.please guide me . yours, sabeti ___ 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] antibody instensity
Hi All, My management team just asked me how feasible a 24 hours IHC lab would be. Are any of you currently running 24 hours? What is the average turn around time for your IHC? We are currently using Ventana platforms and mainly Ventana and Cell Marque antibodies. I do have concern with keeping most of the antibodies at room temp all day, every day. Has anyone experienced a drift in intensity with any antibodies? Any input for or against would be appreciated. Thank you. :-) Melissa Kuhnla Lead Medical Technologist for IHC and FISH Catholic Health Services of Long Island Regional Laboratory Services The information in this e-mail, and any attachments therein, is confidential and for use by the intended addressee only. If this message is received by you in error please do not disseminate or read further. Please reply to the sender that you have received the message in error, then delete the message. Although Catholic Health Services of Long Island attempts to sweep e-mail and attachments for viruses, it does not guarantee that either are virus-free and accepts no liability for any damage sustained as a result of viruses. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Grossing Station Recommendation
Grosslab Senior here as well. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcne...@lmhealth.org www.LMHealth.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sean McBride Sent: Monday, March 21, 2011 2:24 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Grossing Station Recommendation Colleagues, We are looking to replace our current pathology grossing hood with a new station, so I am looking for recommendations. Thanks in advance for all of your wonderful advice. 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, including attachments, is intended for the sole use of the individual and/or entity to whom it is addressed, and contains information from Licking Memorial Health Systems which is confidential or privileged. If you are not the intended recipient, nor authorized to receive for the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this e-mail and attachments is prohibited. If you have received this in error, please advise the sender by reply e-mail and delete the message immediately. You may also contact the LMH Process Improvement Center at 740-348-4641. E-mail transmissions cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The sender therefore does not accept liability for any errors or omissions in the contents of this message, which arise as a result of e-mail transmission. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] how to bleach melanoma cases without bad effect on IHC
hello dear fellows, i have an IHC project on paraffin embedded melanoma cases; some of them being heavily pigmented. i have seen many protocols based on H2O2 to bleach the pigmented ones, but i am not sure about any possible effect on antigen retrieval.please guide me . yours, sabeti ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
