RE: [Histonet] TMA Arrayers
Hello, We have the Pathology Devices Semi automated units. We are happy with them. Our decision to keep with the less automated units are largely due to our work flow. We make TMA's for customers. But the customer has to do all of the work in the design and data entry into out TMAJ Database. I think all of the automated units have data input features that are not compatible with this work flow. Good luck in choosing. Helen L. Fedor Oncology Tissue Services, Manager Johns Hopkins University 411 N. Caroline St Room 310 Basement| Bond St Annex Building Baltimore, MD | 21231 410-614-1660 http://tmalab.jhmi.edu/ -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sally Ann Drew Sent: Thursday, April 30, 2015 1:36 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] TMA Arrayers I was wondering how many people have semi-automated or automated tissue microarray equipment? I am especially interested in those who graduated from manual methods to the automated and how they reviewed and ranked the limited options out there. Thank you! _Sally Sally Ann Drew, MT(ASCP) UWSMPH-Dept. of Pathology TRIP Lab Manager Translational Science BioCore CSC, L5/181 608.265.1093 ___ 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] TMA Arrayers
I was wondering how many people have semi-automated or automated tissue microarray equipment? I am especially interested in those who graduated from manual methods to the automated and how they reviewed and ranked the limited options out there. Thank you! _Sally Sally Ann Drew, MT(ASCP) UWSMPH-Dept. of Pathology TRIP Lab Manager Translational Science BioCore CSC, L5/181 608.265.1093 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Disopsal of containers?
This may sound like a really sophomoric question but I'll ask anyway. How do you dispose of specimen containers from your histo tissues once they are emptied and rinsed? This question came up at work (veterinary diagnostic lab) and I am not aware of a standard or a reference to go to for an answer. Thanks in advance. Carole Johnson Carole Johnson, HT(ASCP)cm New Mexico Department of Agriculture Veterinary Diagnostic Services 505.383.9299 To understand is to stand under, which is to look up, which is a good way to understand Confidentiality Notice: New Mexico has a very broad public records law. Most written communications to or from state employees are public records. Your e-mail communications may therefore be subject to public disclosure. This e-mail, including all attachments is for the sole use of the intended recipients. Any unauthorized review, use, disclosure or distribution is prohibited unless specifically provided under the New Mexico Inspection of Public Records Act. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Question
All, I have a procedure here that call for and I quote 1.25 ml NaOH 10N in 1L of water. I know how to get 1 N, but how do I get 10. Having rarely hd the opportunity to make many Normal solutions ,my brain is not computing. Is it an error? Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-freder...@northwestern.edumailto:b-freder...@northwestern.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Question
Bernice, You have to buy the 10N solution. You can only dilute a given normal solution, you cannot concentrate them. Tim Morken Pathology Site Manager, Parnassus Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bernice Frederick Sent: Thursday, April 30, 2015 12:35 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Question All, I have a procedure here that call for and I quote 1.25 ml NaOH 10N in 1L of water. I know how to get 1 N, but how do I get 10. Having rarely hd the opportunity to make many Normal solutions ,my brain is not computing. Is it an error? Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-freder...@northwestern.edumailto:b-freder...@northwestern.edu ___ 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] ANP.23410
Usually I have done weekly , but in rather high usage places. With more strict downtimes and decontamination for TB or other suspected infectious cases. Joelle Weaver MAOM, HTL (ASCP) QIHC From: dknut...@primecare.org To: histonet@lists.utsouthwestern.edu Date: Thu, 30 Apr 2015 14:20:54 -0500 Subject: [Histonet] ANP.23410 Hi fellow Histonetters - I was wondering if I could get some feedback from my peers on how you are dealing with the CAP standard ANP.23410 on Cryostat Decontamination. It states that this is done at defined intervals appropriate for the institution. The place I just inspected was doing a weekly decontaminating, and a more thorough one quarterly. I would like to know how often are other sites taking apart the entire cryostat chamber for decontamination. Thank you so much for sharing your process. Deanne Knutson Supervisor Anatomic Pathology This email may include confidential and privileged information. If this is not intended for your use, please destroy immediately and contact the sender of the message. This email and attachments contain information that may be confidential or privileged. If you are not the intended recipient, notify the sender at once and delete this message completely from your information system. Further use, disclosure, or copying of information contained in this email is not authorized, and any such action should not be construed as a waiver of privilege or other confidentiality protections. ___ 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] ANP.23410
Hi fellow Histonetters - I was wondering if I could get some feedback from my peers on how you are dealing with the CAP standard ANP.23410 on Cryostat Decontamination. It states that this is done at defined intervals appropriate for the institution. The place I just inspected was doing a weekly decontaminating, and a more thorough one quarterly. I would like to know how often are other sites taking apart the entire cryostat chamber for decontamination. Thank you so much for sharing your process. Deanne Knutson Supervisor Anatomic Pathology This email may include confidential and privileged information. If this is not intended for your use, please destroy immediately and contact the sender of the message. This email and attachments contain information that may be confidential or privileged. If you are not the intended recipient, notify the sender at once and delete this message completely from your information system. Further use, disclosure, or copying of information contained in this email is not authorized, and any such action should not be construed as a waiver of privilege or other confidentiality protections. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
FW: [Histonet] IHC and oven temperature
Yes, I read the Dako IPX educational guides (5th ed) and on page 32: No processes should raise tissue temperature to higher than 60oC as this will cause severe loss of antigenicity that may not be recoverable Unfortunately there is no evidence given or cited that validates this statement. Even though this could be right (and there are several papers that have looked at this), this statement is scientifically weak and we should not cite this as truth. Now I do recommend the Dako reference series to my students, and I have contributed to one of these texts myself (Microscopic control of routine HE - know your histology) but I request my students to continue to question what they read and confirm the scientific validity of the information. Regards, Tony From: Joelle Weaver [joellewea...@hotmail.com] Sent: Saturday, 25 April 2015 5:51 AM To: Tony Henwood (SCHN); WILLIAM DESALVO; Preiszner, Johanna Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] IHC and oven temperature I remember reading that the preffered temperature was about 60 degrees Celsius. I think that this was in the Dako education guides if I'm not mistaken. If that is the case, the citation for the source is probably in that resource available as pdf from their website. Joelle Weaver MAOM, HTL (ASCP) QIHC From: tony.henw...@health.nsw.gov.au To: wdesalvo@outlook.com; preis...@mail.etsu.edu Date: Fri, 24 Apr 2015 09:43:59 + Subject: RE: [Histonet] IHC and oven temperature CC: histonet@lists.utsouthwestern.edu Hi temp drying shown to be a bad idea: Henwood, A., (2005) “Effect of Slide Drying at 80°C on Immunohistochemistry” J Histotechnol 28(1):45-46. Abstract Prolonged high temperature dry heating has been found to be deleterious to the immunohistochemical demonstration of several antigens in formalin-fixed, paraffin- embedded sections. Paraffin sections were dried at 80°C for 7 h and their immunoreactivity was compared with mirror sections dried for 1 h at 60°C. NCL-5D3, CMV, S100, HMB45, and CEA were quite labile to dry overheating whereas AElAE3, HBsAg, HBcAg, HSVII, EMA, chromogranin, and NSE were found to be quite resistant. It is recommended that coated slides (poly-L-lysine or aminopropyltriethoxysilane) and low-temperature drying (60°C) be routinely used for irnmunohistochemistry. From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] on behalf of WILLIAM DESALVO [wdesalvo@outlook.com] Sent: Tuesday, 21 April 2015 1:56 AM To: Preiszner, Johanna Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IHC and oven temperature Dry heat compared to wet heat. Do not dry your slides at high heat. You are removing water trapped between slide and paraffin section. Antigen retrieval is an entirely different process. So not try to combine the two processes Sent from my iPhone On Apr 20, 2015, at 8:48 AM, Preiszner, Johanna preis...@mail.etsu.edu wrote: Hi Netters, is there something wrong with this logic: If the tissue needs 95C for HIER, it's ok to dry the slides in an 82C oven. Of course I'll test it before I try it on real specimens, but maybe someone else already knows the answer... Thanks! Hanna Preiszner ETSU/QCOM ___ 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 Sydney Children's Hospitals Network. This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Sydney Childrens Hospital's Network 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 * 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
[Histonet] RE: IHC billing question
Ouch! -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cartun, Richard Sent: Thursday, April 30, 2015 4:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC billing question Effective January 1, 2015, our LIS team removed all of the CPT 88342 codes for IHC from our CoPath stain dictionary since you couldn't tell whether a Cytokeratin-7 was being performed as an 88341 or as an 88342. Now, as you might have expected, none of the inpatient IHC testing has been accounted for (the outpatient IHC has been billed manually from the pathology report), and they want someone to go back and enter all the CPT codes into the system (hopefully, not me!). Has anyone else encountered this problem? Thanks (I think). 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 (860) 545-2204 Fax 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC billing question
We do all of our IHC billing manually. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of WILLIAM DESALVO Sent: Thursday, April 30, 2015 4:43 PM To: Cartun, Richard Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IHC billing question We have to manually review the IHC billing also and continue to audit. It took billing and IT three months to create the logic to automate billing for a specimen and account for combination of there being the possibility of 88341, 88342 88344 and the proper combinations. Sent from my iPhone On Apr 30, 2015, at 2:34 PM, Cartun, Richard richard.car...@hhchealth.org wrote: Effective January 1, 2015, our LIS team removed all of the CPT 88342 codes for IHC from our CoPath stain dictionary since you couldn't tell whether a Cytokeratin-7 was being performed as an 88341 or as an 88342. Now, as you might have expected, none of the inpatient IHC testing has been accounted for (the outpatient IHC has been billed manually from the pathology report), and they want someone to go back and enter all the CPT codes into the system (hopefully, not me!). Has anyone else encountered this problem? Thanks (I think). 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 (860) 545-2204 Fax 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 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... Frozen section fixation problems
I have been following this with interest both now and in the past. A word of caution about the acetone/ethanol fixation. I did NOT use the acetone/alcohol fixative cold, but at RT (as it was taught to me by an IHC expert). That is a bonus since you don't have to maintain A/A fixative in a refrigerator. It could be that in Brett's hands, A/A at -20C works well so I can't argue with a successful variation for this fixative.A major caveat: A/A is used for rodent CD markers and cannot be used for human CD4 or CD8 as reported by the late Dr. Chris van der Loos. He and I collaborated about frozen section fixatives many times along with trying each other's method. He always had success with 4C acetone in very humid The Netherlands but was careful to air dry the sections overnight in front of a fan.These two human CD markers do not tolerate ethanol consequently, I wouldn't use A/A for any human CD marker work.We have used it exclusively for murine and rat CD markers and Q-fever organisms. A good rule it to have a panel of fixation methods in order to optimize fixation for any given antigen. I do not understand why Patrick has such problems using cold acetone fixation which leads to poor sections.We air dried frozen sections for a minimum of 30 min before A/A fixation. Most of the time, frozen sections were cut and immediately dried at RT for up to 4 hours, then stored in a box containing only one day's worth of staining. The unfixed sections are stored at -80C with a bag of silica gel in the box (25 slide capacity). The slide box can be taken out the night before staining, or even the day of staining with lid on to NOT GET WATER CONDENSATION ON THE SECTION. Water condensation can damage morphology and antigens. I would NEVER use an acetone gradient for fixation since the increase in water could be a cause of the damage. Water is not going to maintain isotonic conditions and prevent damage. If you want to blow away a frozen section after acetone fixation, just rinse with watera sure way to damage the morphology. After acetone fixation only (10 min at 4C), air dry section for 15 min, then go into PBS or TBS. Before fixation, use barrier pen i.e. ImmEdge (vortexed to mix components before drawing around section) from Vector around section, then fix in A/A 10 min @RT and then go immediately from A/A into pure PBS for 3 changes. The 4th change is PBS/0.2% Tween 20 to equilibrate the section for IHC buffer conditions. What I suspect, after so many continued problems, is the snap freezing of the tissue may be done improperly and the damage could be excessive freezing artifact. Something is amiss and it may be BEFORE FIXATION with the acetone. In general, I have found methanol to be a poor fixative for IHC, and should be totally avoided for any CD marker work since it causes protein hydrolysis of the epitope causing weak, poor staining. 4% paraformaldehyde @ 4C without antigen retrieval can give weak staining and antigen retrieval with frozen sections has to be done carefully to maintain delicate sections on the slide.95%, even 100%, ethanol can also result in weak staining. You did not say what epitopes you are trying to preserve and stain for? I don't think the plus charge slides are the culprit since I had labs using acetone fixation of FS on plain glass slides before Plus charge was so popular. Are you sure your PBS or TBS is correctly made? Incorrectly made PBS caused morphology havoc to completely blow away my frozen sections. This led to purchasing Sigma Dulbecco PBS which never gave problems. Maybe you can describe more of what you are doing from the time you receive and snap freeze the tissues, species, etc., including manual or automated staining in order to have other help you chase away these annoying gremlins. Gayle M. Callis HTL/HT/MT(ASCP) Patrick, We do a lot of frozen section IHC work. Years ago Gayle Callis turned me on to fixing in cold acetone:ethanol (3:1) . We keep it at -20C and I fix for 10 min. on the bench then wash in PBS and proceed with the IHC. We do dry slides for at least 30 min before fixing. This has worked well in our hands for many different antibodies. Brett Brett M. Connolly, Ph.D. Principle Scientist, Imaging Dept. Merck Co., Inc. PO Box 4, WP-44K West Point, PA 19486 brett_connolly @t merck.com T- 215-652-2501 F- 215-993-6803 -Original Message- From: histonet-bounces @t lists.utsouthwestern.edu [mailto:histonet-bounces @t lists.utsouthwestern.edu] On Behalf Of Lewis, Patrick Sent: Tuesday, April 28, 2015 5:56 PM To: (Histonet @t lists.utsouthwestern.edu) Subject: [Histonet] Acetone fixation problems with OCT Tissues Hi Everyone, I am still having issues with my IHCs with Acetone fixation. If I fix in 100% Acetone, I get IHC staining, but my tissues are 50-90% destroyed. If I fix
Re: [Histonet] IHC billing question
We have to manually review the IHC billing also and continue to audit. It took billing and IT three months to create the logic to automate billing for a specimen and account for combination of there being the possibility of 88341, 88342 88344 and the proper combinations. Sent from my iPhone On Apr 30, 2015, at 2:34 PM, Cartun, Richard richard.car...@hhchealth.org wrote: Effective January 1, 2015, our LIS team removed all of the CPT 88342 codes for IHC from our CoPath stain dictionary since you couldn't tell whether a Cytokeratin-7 was being performed as an 88341 or as an 88342. Now, as you might have expected, none of the inpatient IHC testing has been accounted for (the outpatient IHC has been billed manually from the pathology report), and they want someone to go back and enter all the CPT codes into the system (hopefully, not me!). Has anyone else encountered this problem? Thanks (I think). 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 (860) 545-2204 Fax 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC billing question
Your Lis should not have done that. If you are using Copath/Cerner, I think they have automated it now according to their most recent newsletter. Currently, I have to manually change the 88342's to 1's It's somewhat of a pain. But, your Lis should have consulted someone before deleting all codes. Your pathologists should have been on top of it as well imho. Unfortunately, someone has to change. It may be too late to bill too for some. Garrey Sent from my iPhone On Apr 30, 2015, at 5:44 PM, Mike Pence mpe...@grhs.net wrote: We do all of our IHC billing manually. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of WILLIAM DESALVO Sent: Thursday, April 30, 2015 4:43 PM To: Cartun, Richard Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IHC billing question We have to manually review the IHC billing also and continue to audit. It took billing and IT three months to create the logic to automate billing for a specimen and account for combination of there being the possibility of 88341, 88342 88344 and the proper combinations. Sent from my iPhone On Apr 30, 2015, at 2:34 PM, Cartun, Richard richard.car...@hhchealth.org wrote: Effective January 1, 2015, our LIS team removed all of the CPT 88342 codes for IHC from our CoPath stain dictionary since you couldn't tell whether a Cytokeratin-7 was being performed as an 88341 or as an 88342. Now, as you might have expected, none of the inpatient IHC testing has been accounted for (the outpatient IHC has been billed manually from the pathology report), and they want someone to go back and enter all the CPT codes into the system (hopefully, not me!). Has anyone else encountered this problem? Thanks (I think). 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 (860) 545-2204 Fax 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 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: FW: [Histonet] IHC and oven temperature
The statement quoted by Tony from the Dako manual cannot be true because many antigens have to be exposed to water at 100C in order to be immunostained - antigen retrieval. Denaturation of a macromolecule by heat increases the number of exposed epitopes, which typically are short amino acid sequences that bind specifically to the Fab segments of antibody molecules. On the other hand, it is easy to believe that 60C would denature antibody molecules enough to damage their binding sites and impair or prevent immunostaining. According to AWP Vermeer and W Norde (2000), the Fab segments of IgG were denatured when the temperature of a solution slightly exceeded 60C. (The Thermal Stability of Immunoglobulin: Unfolding and Aggregation of a Multi-Domain Protein Biophysical Journal 78: 394–404.) They found that further heating denatured the Fc segment, but the changed molecules became entangled and aggregated before denaturation was complete. Microwave heating is sometimes used to accelerate immunostaining, but control of the temperature is critical. For example: ME Boon E Marani (1991) The major importance of temperature data in publications concerning microwave techniques European Journal of Morphology 29: 181–183. John Kiernan London, Canada = = = On 30/04/15, Tony Henwood (SCHN) tony.henw...@health.nsw.gov.au wrote: Yes, I read the Dako IPX educational guides (5th ed) and on page 32: No processes should raise tissue temperature to higher than 60oC as this will cause severe loss of antigenicity that may not be recoverable Unfortunately there is no evidence given or cited that validates this statement. Even though this could be right (and there are several papers that have looked at this), this statement is scientifically weak and we should not cite this as truth. Now I do recommend the Dako reference series to my students, and I have contributed to one of these texts myself (Microscopic control of routine HE - know your histology) but I request my students to continue to question what they read and confirm the scientific validity of the information. Regards, Tony From: Joelle Weaver [joellewea...@hotmail.com] Sent: Saturday, 25 April 2015 5:51 AM To: Tony Henwood (SCHN); WILLIAM DESALVO; Preiszner, Johanna Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] IHC and oven temperature I remember reading that the preferred temperature was about 60 degrees Celsius. I think that this was in the Dako education guides if I'm not mistaken. If that is the case, the citation for the source is probably in that resource available as pdf from their website. Joelle Weaver MAOM, HTL (ASCP) QIHC From: tony.henw...@health.nsw.gov.au To: wdesalvo@outlook.com; preis...@mail.etsu.edu Date: Fri, 24 Apr 2015 09:43:59 + Subject: RE: [Histonet] IHC and oven temperature CC: histonet@lists.utsouthwestern.edu Hi temp drying shown to be a bad idea: Henwood, A., (2005) “Effect of Slide Drying at 80°C on Immunohistochemistry” J Histotechnol 28(1):45-46. Abstract Prolonged high temperature dry heating has been found to be deleterious to the immunohistochemical demonstration of several antigens in formalin-fixed, paraffin- embedded sections. Paraffin sections were dried at 80°C for 7 h and their immunoreactivity was compared with mirror sections dried for 1 h at 60°C. NCL-5D3, CMV, S100, HMB45, and CEA were quite labile to dry overheating whereas AElAE3, HBsAg, HBcAg, HSVII, EMA, chromogranin, and NSE were found to be quite resistant. It is recommended that coated slides (poly-L-lysine or aminopropyltriethoxysilane) and low-temperature drying (60°C) be routinely used for irnmunohistochemistry. From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] on behalf of WILLIAM DESALVO [wdesalvo@outlook.com] Sent: Tuesday, 21 April 2015 1:56 AM To: Preiszner, Johanna Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IHC and oven temperature Dry heat compared to wet heat. Do not dry your slides at high heat. You are removing water trapped between slide and paraffin section. Antigen retrieval is an entirely different process. So not try to combine the two processes Sent from my iPhone On Apr 20, 2015, at 8:48 AM, Preiszner, Johanna preis...@mail.etsu.edu wrote: Hi Netters, is there something wrong with this logic: If the tissue needs 95C for HIER, it's ok to dry the slides in an 82C oven. Of course I'll test it before I try it on real specimens, but maybe someone else already knows the answer... Thanks! Hanna Preiszner ETSU/QCOM ___ Histonet mailing list Histonet@lists.utsouthwestern.edu
[Histonet] (no subject)
Hi, I was hoping someone can help me with tissue falling off the slides. I have tried regular slides with and without adhesive in the water bath. Charged slides with and without adhesive in the water bath. I have not changed the type of slides I’m using. All the chemicals are fresh in the processor and the stain line, as well as the paraffin in the processor. It is the worst on needle bx ( prostate and breast ). I am SO frustrated, any help would be greatly appreciated! Carolyn Sent from Windows Mail___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] (no subject)
How long do you bake slides for before staining, at what temperature? Does your stainer use agitation? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Carolyn Nelson Sent: Thursday, April 30, 2015 11:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hi, I was hoping someone can help me with tissue falling off the slides. I have tried regular slides with and without adhesive in the water bath. Charged slides with and without adhesive in the water bath. I have not changed the type of slides I’m using. All the chemicals are fresh in the processor and the stain line, as well as the paraffin in the processor. It is the worst on needle bx ( prostate and breast ). I am SO frustrated, any help would be greatly appreciated! Carolyn Sent from Windows Mail -This e-mail and any attachments may contain CONFIDENTIAL information, including PROTECTED HEALTH INFORMATION. If you are not the intended recipient, any use or disclosure of this information is STRICTLY PROHIBITED; you are requested to delete this e-mail and any attachments, notify the sender immediately, and notify the LabCorp Privacy Officer at privacyoffi...@labcorp.com or call (877) 23-HIPAA / (877) 234-4722. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
