Connection: In your opinion, what is the biggest hurdle for standardization? Probably education in laboratory practices. Most of the pathologists are very well trained in diagnosis, but lack bench training and training in laboratory practices. Also, costs associated with training is an important issue in Latin America. Connection: United Kingdom National External Quality Assessment Service (NEQAS) helps to ensure that clinical laboratory test results are accurate, reliable and comparable wherever they are produced. Similarly, NordiQC - an independent, scientific, non-profit organization, is dedicated to the quality assessment of IHC in Europe. What is the equivalent of NEQAS and NordiQC in Latin America? I agree that we need a universal fixative. In our hospital/laboratory, we have used buffered formalin (formaldehyde: 37% diluted 1:10 with distilled water). Regarding the use of other fixatives, I do not recommend them. Most of our knowledge in biomarkers comes from papers using formalin, and I think that the use of other fixatives can introduce a bias in interpretation. Only in special situations, and not in routine material, I can admit the use of other fixatives. 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 Latin America? Although some fixatives give us better morphological details, it should be used in addition to formalin solution (a second fragment). For example, in Latin America there are some selected biopsies such as lymph nodes and renal specimens that can be treated in B5 fixative or Bouins solution for fixation, respectively. The choice of the second fragment rests with the pathologist. 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; (4): 149­153). What guidelines are followed in Latin America for HER2 testing? Most of the well-funded cancer centers in Latin America adopt the ASCO/CAP Guidelines. However, in smaller hospitals, there are no guidelines or any form of standardization associated because of the cost issues. There are also some difficulties in smaller hospitals to introduce HER2 tests - mainly because of the costs. 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 de-cloaking devices). However, the variability of results which are 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 standardisation of antigen , retrieval in Immunohistochemistry using commercially available equipment. Immunocytochemistry 2006; 13-17). What protocol do you follow for antigen retrieval? Do the protocols meet the standards required by UK NEQAS, NordiQC, United States College of American Pathologists (CAP)? Or do you have your own standards in Latin America? step is The firstcourses education - in the form of and workshops about standardization. There is nothing equivalent to NEQAS or NordiQC in Brazil. A few years ago, The Brazilian Society of Pathology had tried a small pilot program on quality control, but it never materialized. Connection: How would you like to standardize IHC in Latin America? The first step is education ­ in the form of courses and workshops about standardization. The second step is to make laboratory managers and hospital administrators aware of the importance of investing funds in IHC laboratories; especially when standardization can save thousands of dollars in diagnosis and antibody-based therapeutics for oncology indications. Connection: How well is the prefixation process standardized? Is it possible to standardize this process? I know of very few hospitals in Latin America that cares about the prefixation process. Most of them use acid formalin, and they are no good at indicating the time of surgery or the time of pathological process and/or other pre-analytical aspects. However, in our hospital we have standardized the prefixation/pre-analytical process since 1997 and, therefore, we are able to control this. 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? Connection 2008 | 10 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