[Histonet] Rabbit anti-Galanin (human)
Hi, I wonder if any one out there working with Peninsula Rabbit anti-Galanin (human)?, I am planing to use it on the rat nervous system tissue which is new work for my lab. I will be appreciate if you can share your great protocol with me :-) Cheers, Leila ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: IHC antibody optimizing validating
I have not read the entire document yet. What do they say about using tissues that have been fixed and processed elsewhere? Richard Richard W. Cartun, MS, PhD Director, Histology Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 972-1596 Office (860) 545-2204 Fax richard.car...@hhchealth.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Terri Braud Sent: Tuesday, March 25, 2014 3:35 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: IHC antibody optimizing validating CAP is very clear that in order to validate a new antibody, that: once the stain has been optimized, that for a well characterized antibody with a limited spectrum of antigenic targets, like Chromogranin or PSA, the validation can be limited. A panel of 10 positive and 10 negative neoplasms would be sufficient in this setting. For an antibody that is not well characterized and/or has a wide range of reported reactivity, a more extensive validation is necessary. The number of tissues tested should in the circumstance be large enough to determine whether the staining profile matches that previously described. An exception to the above requirements is that studies nay not be feasible for antigens such as ALK that are only seen in rare tumors. Thus sayeth CAP. And if you're like me, I am not digging through all my cases to try to come up with 30-40 neoplasms for each antibody, so I just order Tissue Micro Arrays with the neoplasms I need. 20 positive and 20 negative neoplastic tissues on one slide for easy staining and validation. The money you save on reagents to stain one little slide, more than makes up for the cost of the slide. I hope this helps. Terri L. Braud, HT(ASCP) Anatomic Pathology Supervisor Holy Redeemer Hospital Laboratory 1648 Huntingdon Pike Meadowbrook, PA 19046 Ph: 215-938-3676 Fax: 215-938-3874 Today's Topics: 8. IHC antibody optimizing validating (Davis, Cassie) Message: 8 Date: Tue, 25 Mar 2014 09:51:07 -0400 From: Davis, Cassie cda...@che-east.org Subject: [Histonet] IHC antibody optimizing validating Will you help me? I understand we are to use the known positives controls that the manufactures' recommends in the package insert when optimizing the stains, but I need to know what is your general procedure for optimizing (how many different staining protocols do you test) and validating a new antibody (how many different or known positive and negative tissues do you test [predictive markers I understand are 20])? Cassandra Davis cda...@che-east.org 302-575-8095 * - CONFIDENTIALITY NOTICE: This E-Mail is intended only for the use of the individual or entity to which it was sent. It may contain information that is privileged and/or confidential, and the use or disclosure of such information may also be restricted under applicable federal and state law. If you received this communication in error, please do not distribute any part of it or retain any copies, and delete the original E-Mail. Please notify the sender of any error by E-Mail. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, or an employee or agent responsible for delivering the message to the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message, including any attachments. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: IHC antibody optimizing validating
Fortunately, they say nothing at all because if that were the case, they would no longer be able to peddle their Proficiency Programs for IHC, since those too, are fixed and processed elsewhere. Terri L. Braud, HT(ASCP) Anatomic Pathology Supervisor Holy Redeemer Hospital Laboratory 1648 Huntingdon Pike Meadowbrook, PA 19046 Ph: 215-938-3676 Fax: 215-938-3874 -Original Message- From: Cartun, Richard [mailto:richard.car...@hhchealth.org] Sent: Wednesday, March 26, 2014 11:47 AM To: Terri Braud; histonet@lists.utsouthwestern.edu Subject: RE: IHC antibody optimizing validating I have not read the entire document yet. What do they say about using tissues that have been fixed and processed elsewhere? Richard Richard W. Cartun, MS, PhD Director, Histology Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 972-1596 Office (860) 545-2204 Fax richard.car...@hhchealth.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Terri Braud Sent: Tuesday, March 25, 2014 3:35 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: IHC antibody optimizing validating CAP is very clear that in order to validate a new antibody, that: once the stain has been optimized, that for a well characterized antibody with a limited spectrum of antigenic targets, like Chromogranin or PSA, the validation can be limited. A panel of 10 positive and 10 negative neoplasms would be sufficient in this setting. For an antibody that is not well characterized and/or has a wide range of reported reactivity, a more extensive validation is necessary. The number of tissues tested should in the circumstance be large enough to determine whether the staining profile matches that previously described. An exception to the above requirements is that studies nay not be feasible for antigens such as ALK that are only seen in rare tumors. Thus sayeth CAP. And if you're like me, I am not digging through all my cases to try to come up with 30-40 neoplasms for each antibody, so I just order Tissue Micro Arrays with the neoplasms I need. 20 positive and 20 negative neoplastic tissues on one slide for easy staining and validation. The money you save on reagents to stain one little slide, more than makes up for the cost of the slide. I hope this helps. Terri L. Braud, HT(ASCP) - CONFIDENTIALITY NOTICE: This E-Mail is intended only for the use of the individual or entity to which it was sent. It may contain information that is privileged and/or confidential, and the use or disclosure of such information may also be restricted under applicable federal and state law. If you received this communication in error, please do not distribute any part of it or retain any copies, and delete the original E-Mail. Please notify the sender of any error by E-Mail. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: IHC antibody optimizing validating
only 1/2 way through reading the article, but Terri makes a good point about the proficiency testing. Joelle Weaver MAOM, HTL (ASCP) QIHC Date: Wed, 26 Mar 2014 12:48:46 -0400 From: tbr...@holyredeemer.com To: richard.car...@hhchealth.org; histonet@lists.utsouthwestern.edu CC: Subject: [Histonet] RE: IHC antibody optimizing validating Fortunately, they say nothing at all because if that were the case, they would no longer be able to peddle their Proficiency Programs for IHC, since those too, are fixed and processed elsewhere. Terri L. Braud, HT(ASCP) Anatomic Pathology Supervisor Holy Redeemer Hospital Laboratory 1648 Huntingdon Pike Meadowbrook, PA 19046 Ph: 215-938-3676 Fax: 215-938-3874 -Original Message- From: Cartun, Richard [mailto:richard.car...@hhchealth.org] Sent: Wednesday, March 26, 2014 11:47 AM To: Terri Braud; histonet@lists.utsouthwestern.edu Subject: RE: IHC antibody optimizing validating I have not read the entire document yet. What do they say about using tissues that have been fixed and processed elsewhere? Richard Richard W. Cartun, MS, PhD Director, Histology Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 972-1596 Office (860) 545-2204 Fax richard.car...@hhchealth.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Terri Braud Sent: Tuesday, March 25, 2014 3:35 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: IHC antibody optimizing validating CAP is very clear that in order to validate a new antibody, that: once the stain has been optimized, that for a well characterized antibody with a limited spectrum of antigenic targets, like Chromogranin or PSA, the validation can be limited. A panel of 10 positive and 10 negative neoplasms would be sufficient in this setting. For an antibody that is not well characterized and/or has a wide range of reported reactivity, a more extensive validation is necessary. The number of tissues tested should in the circumstance be large enough to determine whether the staining profile matches that previously described. An exception to the above requirements is that studies nay not be feasible for antigens such as ALK that are only seen in rare tumors. Thus sayeth CAP. And if you're like me, I am not digging through all my cases to try to come up with 30-40 neoplasms for each antibody, so I just order Tissue Micro Arrays with the neoplasms I need. 20 positive and 20 negative neoplastic tissues on one slide for easy staining and validation. The money you save on reagents to stain one little slide, more than makes up for the cost of the slide. I hope this helps. Terri L. Braud, HT(ASCP) - CONFIDENTIALITY NOTICE: This E-Mail is intended only for the use of the individual or entity to which it was sent. It may contain information that is privileged and/or confidential, and the use or disclosure of such information may also be restricted under applicable federal and state law. If you received this communication in error, please do not distribute any part of it or retain any copies, and delete the original E-Mail. Please notify the sender of any error by E-Mail. Thank you for your cooperation. ___ 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] CAP Question, New Reagent Lot Confirmation of Acceptability
The above checklist item, ANP.22760, states The performance of new lots of antibody and detection system reagents is compared with old lots before or concurrently with being placed into service. Neither the question, the note below it, nor the evidence of compliance states that a pathologist has to perform the comparison. What are other labs doing? Are the histotechs signing off on new lots? Laurie Colbert, HT (ASCP) Histology Supervisor PATH MD 8158 Beverly Blvd. Los Angeles, CA 90048 (323) 648-3214 direct (424) 245-7284 main lab The information contained in this transmission may contain privileged and confidential information, including patient information protected by federal and state privacy laws. It is intended only for the use of the person(s) named above. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution, or duplication of this communication is strictly prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC on animals
Hi: Can IHC validated for humans be performed on animal tissues without any protocol changes? Thanks, Lester J. Raff, MD MBA UroPartners Medical Director Of Laboratory 2225 Enterprise Dr. Suite 2511 Westchester, Il 60154 Tel: 708-486-0076 Fax: 708-492-0203 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Frozen sectioning whole rat eyes
Hello Histoland! A colleague of mine is trying to section unfixed whole rat eyes in OCT. She is getting a lot of tearing at the top of her section. Lens and optic nerve on the horizontal plane. Any suggestions, recommendations would be great! Jackie ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC on animals
Dr. Raff, You will need to do trial runs to first determine if the antibody can detect the specific epitope in animal tissues (epitopes aren't always conserved between species), and then do your own optimization to determine the best dilution and tissue pretreatment for that particular species. They can vary by species, and don't always coincide with the recommendations on the antibody data sheet provided by the vendor. -- Jan Shivers Senior Scientist IHC/Histology Section Head Pathology Teaching Program Veterinary Diagnostic Laboratory University of Minnesota 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive...@umn.edu On Wed, Mar 26, 2014 at 3:03 PM, Lester Raff MD lr...@uropartners.comwrote: Hi: Can IHC validated for humans be performed on animal tissues without any protocol changes? Thanks, Lester J. Raff, MD MBA UroPartners Medical Director Of Laboratory 2225 Enterprise Dr. Suite 2511 Westchester, Il 60154 Tel: 708-486-0076 Fax: 708-492-0203 ___ 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: IHC on animals
Lester I would not, first of all not all antibodies that work in human will cross react with a particular species of animal. Another thing to consider is that most human based detection systems consist of dual link reagents, meaning that they work on both mouse and rabbit primaries these detection system are not ideal for working with animals and depending upon the species that you are working with may cause some problems in with background staining due to the secondary antibody or polymer binding to endogenous IgG in the samples. You can initially pilot your protocol on a particular species to see what happens but I would not take the chance and run a bunch of slides thinking that the protocol that you have in place will work in the particular species of animal you need to stain. Good Luck Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lester Raff MD Sent: Wednesday, March 26, 2014 2:03 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on animals Hi: Can IHC validated for humans be performed on animal tissues without any protocol changes? Thanks, Lester J. Raff, MD MBA UroPartners Medical Director Of Laboratory 2225 Enterprise Dr. Suite 2511 Westchester, Il 60154 Tel: 708-486-0076 Fax: 708-492-0203 ___ 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] Body Release and Cleanin in Morgue
Hello to all in histo-land. Who takes care of releasing the body to the funeral home and who does the cleaning of the seals and the molds on the drawers for the body storage? Is lab some how involved in this process? Allison Scott HT(ASCP) Supervisor, Histology Lab LBJ Hospital Harris Health System Office: 713-566-2148 Lab: 713-566-5287 CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 (HIPAA), PL 104-191; 45 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: IHC on animals
Lester, Probably not in most cases but it depends upon what the antibody is, whether as a an antihuman ab it also cross reacts to the species you are interested in and what detection reagents you are using. For instance if you have determined a rabbit anti human antibody also cross reacts in mouse tissue, you could use a detection such as for the Leica Bond by leaving out the link (they call it the post primary but it is rabbit anti mouse IgG to link mouse antibodies to the goat anti rabbit labeled polymer second step), since your ab is rabbit it would link directly to the anti rab labeled polymer and the anti mouse link which would bind non specifically to the mouse endogenous Ig can be avoided. You would have to do a lot of research to first determine if the anti human rabbit antibody you want to use (it should not be made in a mouse) cross reacts to mouse tissue. The species you are wanting to label is also very important, this system would only work for anti human rab abs that also react in mouse, rat or any other species except rabbit and goat because those are the species the detection is made in or against. We have research versions of the Leica Bond instruments which allows us to manipulate the instrument and detection for these purposes including replacing the rab anti ms link with another link, I am not sure if the pure clinical versions of the Bond allow this. Cheers, Patsy Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting 40864 E. Arkansas Ave Bennett, CO 80102 H 303-644-4538 C 720-281-5406 prueg...@hotmail.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Wednesday, March 26, 2014 2:20 PM To: Lester Raff MD; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: IHC on animals Lester I would not, first of all not all antibodies that work in human will cross react with a particular species of animal. Another thing to consider is that most human based detection systems consist of dual link reagents, meaning that they work on both mouse and rabbit primaries these detection system are not ideal for working with animals and depending upon the species that you are working with may cause some problems in with background staining due to the secondary antibody or polymer binding to endogenous IgG in the samples. You can initially pilot your protocol on a particular species to see what happens but I would not take the chance and run a bunch of slides thinking that the protocol that you have in place will work in the particular species of animal you need to stain. Good Luck Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lester Raff MD Sent: Wednesday, March 26, 2014 2:03 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on animals Hi: Can IHC validated for humans be performed on animal tissues without any protocol changes? Thanks, Lester J. Raff, MD MBA UroPartners Medical Director Of Laboratory 2225 Enterprise Dr. Suite 2511 Westchester, Il 60154 Tel: 708-486-0076 Fax: 708-492-0203 ___ 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
