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 India? Though I agree that cytological features are well-outlined by above fixatives they are not economical and practical for general usage. I am not aware of any laboratory in India using these fixatives. Connection: The American Society of Clinical Oncology (ASCO) and the College of American Pathologists (CAP) convened an expert panel to develop a US guideline to improve the accuracy of HER2 testing in invasive breast cancer. The ASCO/CAP Guideline is also followed by Canadian pathologists with some modifications (Hanna W, et al. Curr Oncol 2007 August;14(4):149­153). Can you explain the guideline followed in India for HER2 testing? We follow the same ASCO GGuideline in India, but as the HerceptTestTM is expensive and all of our patients cannot afford it routinely, we use cerbB-2 Ab (Immunotech, Marseille, France) to screen for the score 2 or 3 cases. These results are validated by either HercepTestTM (DAKO) or FISH, if the patient is to be treated with imatinib. Connection: How about estrogen and progesterone receptor biomarker tests? We use DAKO Antibodies for ER 1d5 and PR and have submitted a paper for publication on our experiences in the standardization of these antibodies. As of now we report any degree of positivity for these markers and score them using a modified histochemical score. Connection: Antigen retrieval is essential in immunohistochemistry (IHC) in order to restore epitopes. There are a large number of devices, which can be used to achieve this aim (microwaves, rice steamers, pressure cookers, autoclaves and commercial decloaking devices). However, the variability of results achieved, not only from one device to another, but also between and within runs using the same device, can be vast (Gray DS, Selbie D, Cooper RF et , al. Simultaneous de-waxing and standardization of antigen retrieval in Immunohistochemistry using commercially available equipment. Immunocytochemistry 2006;4:13-17). What protocol do you follow for antigen retrieval? Do the protocols meet the standards required by UK NEQAS, NordiQC and United States College of American Pathologists (CAP)? Or do you have your own standards in India? Before an antibody is released for use it is standardized and whatever antigen retrieval works best as per international norms. We prefer the pressure-cooking method for most antibodies (especially ER/PR) as microwave does not always ensure uniform retrieval. We are also shifting to the polymer technology for better results. In many tissues, endogenous biotin gives a background staining that interferes with interpretation of the result. This non-biotin-based technology has greater sensitivity and eliminates the need for avidin/ biotin blocking while maintaining the specificity of the reaction (cf. Sabattini E, Bisgaard K, Ascani S, Poggi S, Piccioli M, Ceccarelli C, Pileri F, Fraternali-Orcionig, Pileri S A. The EnVisionTM+ system: A new immunohistochemical method for diagnostics and research. Critical comparison with the APAAP, ChemMateTM, CSA, LABC, and SABC Techniques. J Clin Pathol 1998;51:506­511). It is known that different permutation and combinations work in a given technical situation. E.g. it has been observed that a combination of 6F11/EnVisionTM did not work as good as EDTA for ER/PR. (cf. Vassallo, J Pinto, G A Alvarenga, M Zeferino, L.; Chagas, C A; Metze K Comparison of Immunoexpression of 2 Antibodies for Estrogen Receptors (1D5 and 6F11) in Breast Carcinomas Using Different Antigen Retrieval and Detection Methods. Applied Immunohistochemistry & Molecular Morphology 12(2):177-182, June 2004). the larger Iinwould say at par withlaboratories India are IHC standards in the world ... Connection: How would you rate European (UK NEQAS, NordiQC) and US IHC standards to Indian IHC standards? I would say the larger laboratorie in India are at par with IHC standards in the world, but we need to stamp an approval through international QA programs. Our institute has participated in several clinical trials based on hormone receptor/Her2neu expression in breast cancer, and the results were comparable. Connection: Are Indian IHC standards followed anywhere else in the world (e.g. Sri Lanka, Nepal)? I do not know. 21 | Connection 2008 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. Denver, CO NSH workshop attend New Premium Quality Concentrated Antibodies New Products Your current and future needs drive the c Targeting Colorectal Adenocarcinomas, Anti-TIMP-1 and Anti-Villin Monoclonal Mouse Anti-Human Tissue Dakos FLEX Ready-to-Use Concept Enhance performance of your laboratory by running FLEX Ready-to-Use 2008 Product Catalog Available Now! The catalog features more than 170 new products, including the A