Connection: United Kingdom National External Quality Assessment Service (UK NEQAS) helps to ensure that clinical laboratory test results are accurate, reliable and comparable wherever they are produced. Similarly, Nordic Immunohistochemical Quality Control (NordiQC) - an independent, scientific, non-profit organization is dedicated in the quality assessment of IHC in Northern Europe. What is the equivalent of UK NEQAS and NordiQC in Canada? In the US, the closest thing I can relate to UK NEQAS and NordiQC would be the National Society For Histotechnology-endorsed College of American Pathologists (CAP) HistoQIP http://www.nsh.org/organizations.php3?action=printContentItem&orgid=111&typeID=1206& itemID=18021. This educational program is designed to evaluate histologic technique, tissue fixation and processing, embedding, microtomy, staining and coverslipping of slides. As part of this program, immunohistochemistry slides are similarly evaluated. Connection: Internal quality procedures are essential for the reproducibility of the IHC performance in the individual laboratory. In contrast, external quality assessment (EQA) system retrospectively and objectively compares staining results from many laboratories by means of an external agency. EQA system allows the identification of insufficient stains and inappropriate protocols, as well as identification of possible interpretation problems. What is the name of your EQA system? As stated previously, MGH Surgical Pathology uses HistoQIP from CAP. Each bi-annual test cohort requires the submission of one immunohistochemistry slide for evaluation. Connection: What are the main functions of the HistoQIP program? The main function of the HistoQIP program is to evaluate the entirety of the produced slide. Slides are requested from various techniques and are evaluated by several experts. Their evaluation encompasses section preparation, presentation, and staining integrity. The immunohistochemistry slides are evaluated for the same criteria and additionally for chromogenic intensity, fixation and sectioning artifacts. Information submitted with slides includes the instruments used during processing, including the tissue processor, microtome and blade type used for cutting the sections, type and duration of fixation, slide thickness, and antibody used. Connection: How would you like to standardize IHC in the United States? I would like to see standardization of immunohistochemistry include recommended formalin fixation times, standard thicknesses for slide preparation. IHC protocols should be optimized for formalin-fixed tissue to include the time the specimen is collected until the time the tissue is placed in ten percent neutral-buffered formalin (NBF). Standard dilutions and titrations of antibodies should likewise be adhered to whenever possible. Connection: How well is the prefixation process standardized? Is it possible to standardize this process? Currently, the prefixation processes are not standardized well. The duration of time from specimen collection until its placement in formalin is rarely monitored or documented. Occasionally, this data is collected on frozen section cases. Similarly, once the tissue is placed in formalin, the duration of time until the specimen is sectioned is often times noticeably absent. The age of a specimen that has previously been embedded in paraffin also has a great bearing on the antigenicity of that specimen. The longer the exposed tissue has been open to the air, the weaker the antigenicity. The only way standardization can take place is to monitor and report these times and optimize staining based on these factors. Connection: The Members of the Ad-Hoc Committee on Immunohistochemistry Standardization (Appl Immunohistochem Mol Morphol 2007; 15:124­133) strongly discourage non-formalin fixatives and encourage formaldehyde as the fixative of choice. What fixatives are used in your laboratory, and at what concentration? Do you recommend the use of any other fixatives and why? MGH Pathology uses the traditional ten percent neutral-buffered formalin (NBF) as the fixation of choice. However, various other fixatives are used in certain subspecialties, including Pen-Fix and B-Plus for hematopathology and lymphoma-leukemia studies. Connection: Is it true that a particular histology feature may be better demonstrated by other fixatives, such as Glyo-Fixx for nuclear features and lymphocyte appearance and Omnifix for cytoplasmic detail (Arch Pathol Lab Med. 2005;-129: 502-506)? Would you recommend these fixatives in the future? Are these fixatives used in the United States? As I understand it, while nuclear features and lymphocyte appearance are excellent with Glyo-Fixx, there is also a loss of low molecular-weight antigens when using this fixative. When processing tissue for microscopy, the objective (no pun intended) is to utilize reagents and protocols that will provide a reliable and reproducible fixation without interfering with the need to perform subsequent testing. Knowing that the use of a particular fixative will alter the antigenicity of a specimen and limit the ability to perform IHC at a later time, leads me to resist the temptation to gain immediate satisfaction (nicely stained nuclei) at the expense of potentially diagnostic methodologies in the future. Connection 2008 | 28 April 2008, VOL 11 Connection IHC STANDARDIZATION AROUND THE WORLD In this issue Q&A with Patho Contents APRIL 2008, VOL 11 2 3 4 Editorial George L. Kumar, PhD Featured Laboratory The Osamura Editorial Immunohistochemistry (IHC) Standardization Around the World George L. Kumar, PhD Managing Featured Laboratory The Osamura Laboratory Laboratory headed by Prof. Robert Yoshiyuki Osamura, MD, Q&A Ask the Experts: On Immunohistochemistry (IHC) Standardization Professor Leong is Medical Di time interval between removal of tissue and immersion in fixative, the temperature of tissue storage Connection: How would you like to standardize IHC in Australia? As discussed above, standardization Connection: Antigen retrieval is essential in immunohistochemistry (IHC) in order to restore epitope Connection: Is a universal image analysis system feasible? No, not until we standardize the all impo Q&A Fernando Soares, MD, PhD University of São Paulo, São Paulo, Brazil Dr. Fernando Soares is F Connection: In your opinion, what is the biggest hurdle for standardization? Probably education in l There are no standards in Latin America. We always follow the manufacturers protocol during our firs Q&A Bryan R. Hewlett, ART, MLT Consultant Technologist to the Anatomic Pathology EQA program of There is no standardization of AR steps circumstances, it would be impossible toand, undera the pre Connection: How would you rate European (UK NEQAS, NordiQC) and US IHC standards to Canadian IHC sta Q&A Prof. Chen Jie Prof. Cui Quancai Peking Union Mediacl College Hospital, Beijing, China Prof. Connection: According to Goldstein et al. Appl Immunohistochem Mol Morphol 2007;15:124­133 Immunohis Connection: Is it true that a particular histology feature may be better demonstrated by other fixat Connection: What constitutes standardization of image analysis as applied to immunohistochemistry (I Q&A Dr. Tanuja Manjanath Shet Dr. Vani Parmar Tata Memorial Hospital, Mumbai, India Tanuja Manj ... the biggest hurdle in India is suboptimal fixation and processing of tissues. Though I agree th Connection: Is it true that a particular histology feature may be better demonstrated by other fixat Connection: Can you comment on the internal and external quality control (EQC) procedures followed i Q&A Prof. Robert Yoshiyuki Osamura Department of Pathology, Tokai University School of Medicine Connection: In your opinion, what is the biggest hurdle for standardization? The biggest hurdle for the standardization of image ... appropriate for pre-screeninganalysis is of the staining quality. Connection: Why is standardization of image analysis in diagnostic pathology important? Because the Q&A James F. Happel, DLM (ASCP), HTL Technical Director of Surgical Pathology, Research and Deve Connection: United Kingdom National External Quality Assessment Service (UK NEQAS) helps to ensure t Connection: The American Society of Clinical Oncology (ASCO) and the College of American Pathologist would recommend that standardization Ibegin with identifying a reliable and trustworthy source ... Interview Immunohistochemistry for Oestrogen and Progesterone Receptors Dr. Andrew Lee Consultant H Connection: What is the difference between the H score and the Allred score? Which is better? What d Connection: Can you comment on the burden in the laboratory, if one changes from a current ER/PR ass Opinion & Interview IHC Standardization: A Dako Perspective Dr. Ole Rasmussen R&D Director, Connection: Dako has developed Readyto-Use Antibodies for in vitro diagnostic applications. How is t Articles UK NEQAS-ICC & ISH: Historical perspective, current role, future directions Andy Dodso UK NEQAS-ICC in the 1990s In his first year as Scheme Organiser, Keith oversaw the transition to sub The application could be argued to represent a field change in terms of the rigour with which the an Assessment teams consist of four assessors, who view slides around a multi-headed microscope and sco The archive which UK NEQAS holds, both in terms of stained slides and methodological data, must sure For Immunocytochemistry and FISH RESULT: RUN 80L SLIDE: NEQAS Laboratory No: XXX Mr. A. Scientist De Figure 6. Feedback on results has always been given high priority, and for many years this has been a b c d Figure 7. The antigen chosen by Gerry Reynolds for his very first assessment run was kap Bibliography Selected UK NEQAS-ICC & ISH papers. Ibrahim M, Dodson AR, Barnett S, Fish D, Jasani Articles Nordic Immunohistochemical Quality Control (NordiQC) An Organization for External Quality A parameters (i.e. results interpretation and reporting) (4, 5). In an EQA setting, by circulating ser CD79a (Fig. 2) Among 112 laboratories submitting stains in the latest run, most used mAb clone JCB11 References 1. Rhodes A, Jasani B, Barnes DM, Bobrow LG, Miller KD. Reliability of immuno-histochemic Fig. 2. CD79a A. Optimal CD79a staining of the tonsil using the monoclonal antibody (mAb) clone JCB Standardization of Ki-67 Immunohistochemical Staining for Diagnosing Grade of Gastrointestinal Strom was conducted in 49 GIST cases. The concordance rate for the evaluation results at three laboratorie CB pH6 a b c d TE pH9 e f g h Autoclave 121° C/10 min Water bath 95° C/40 min Microwave 50 Table 1. Correlation between NCC and STD methods R2=0.9483 Categories of proportion NCC Method 3 Opinions Importance of Standardization for Predictive Prognosis David J. Dabbs, MD Chief of Pathol is documented and serves as a surrogate marker for the initial exposure to formalin. Since the first Opinions The New Era for ER and PRIts time to Standardize! Dr. Ian Ellis, B.Med.Sci. BM, MS, FRCpa et al 2001). The main reason for false-negative results is due to inefficient heat-induced epitope ( Standardization of HER2 TestingInconsistency Raises Questions Opinions Sunil S. Badve, MD, FRCPath rence seen in these trials is in the order of 50%. This is the major reason for all the excitement a which now requires expression of HER2 by at least 30% of tumor cells (instead of 10%). It has also r IHC CONSENSUS MEETING, JANUARY 27 2008, SANTA BARBARA, CA, USA , IHC CONSENSUS MEETING, JANUARY 27, Richard Cartun, PhD, Sunil Badve, MD Jon Askaa, PhD Søren Nielsen, HT, CT, Mogens Vyberg, MD Elizab Dako Abstracts Abstracts presented at the 30th Annual San Antonio Breast Cancer Symposium December Dako Abstracts Amplification of ESR1 may predict resistance to adjuvant tamoxifen in postmenopausal Dako Abstracts Abstracts presented at the United States and Canadian Academy of Pathology (USCAP) A Dako Abstracts Metastatic Pancreatic Endocrine Tumors in the Liver Express KOC Briones AJ, Bourne P Dako Abstracts Merkel Cell Carcinomas Express K Homology Domain Containing Protein Overexpressed in Dako Abstracts Immunohistochemical Analysis of KOC/IMP3 in Malignant Pleural Mesothelioma Xu H, Sim Dako Abstracts KOC, TTF-1 and CDX2 Discriminate Small-Cell Carcinoma from Carcinoid and Pancreatic Dako Publications Publications Co-authored by Dako: In Press Li L, Xu H, Spaulding B, Cheng L, Si Dako Meetings 2007 - National Society for Histotechnology Meeting. 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