Improving Immunohistochemistry

Friday, 20 April 2007

Improving Immunohistochemistry
Friday, 20 April 2007 09:00 - 17:00

Imperial College London
Drewe Lecture Theatre
Reynolds Building
St. Dunstan Road
W6 8RP
United Kingdom

Map and Directions

Improving immunohistochemistry brings together clinical, academic and commercial practitioners for an annual update of latest developments in the practice of this and allied technologies. The emphasis of this year’s meeting will be on getting the best from the technology.

Meeting's Chair:  Dr Tony Warford - Head of Immunohistochemistry, AstraZeneca Global R&D, Shanghai, China

9:15 - 9:45 Registration

9:45 ­- 10:00 Introduction by the Chair:
Dr Tony Warford, Wellcome Trust Sanger Institute, Cambridge, UK

10:00 ­- 10:30 The use of glycol methracylate embedding for immunohistochemistry
Dr Susan Wilson, Head of the Histochemistry Research Unit, Infection, Inflammation &
Repair, Southampton General Hospital, UK
Embedding of tissue samples into glycol methacrylate (GMA) resin for immunohistochemical studies (IHC) offers numerous advantages over frozen and paraffin preparations. The use of a non-cross linking fixative (acetone), together with processing at low temperature, ensures good antigen preservation; the tissue morphology is excellent; thin sections (1-2mm) can be cut allowing for several sections to be made through one cell enabling co-localisation studies and a large number of sections to be cut from one sample and embedded tissue can be stored relatively long-term. However, this is a specialised technique and is not applicable to all tissue types or laboratories.

10:30 ­- 11:00 Development of Standardised Control Materials for Immunocytochemistry
Dr. Anthony Rhodes, Senior Lecturer in Cellular Pathology, Centre for Research in
Biomedicine, UWE Bristol, UK
Therapies targeted to over-expressed tumour biomarkers require sensitive and reproducible laboratory assays in order to identify those patients most likely to benefit from this type of treatment. However, sophisticated biological reference materials are also required to closely monitor immunocytochemical assay sensitivity and prevent false negative and false positive results. Cell lines with a graded series of biomarker expression, grown under stringently quality assured conditions to guard against batch-to-batch variation in protein expression, represent the most sustainable and appropriate source of reference material. The talk will emphasise the current and future potential of cell lines for this application.

11:0011:30 Mid-morning break

11.30 ­- 11.45 Affinity is not the only factor
Dr Mark Rees, Vision BioSystems Ltd, UK
Recently there has been a broad interest into the effect of antibody affinity on the quality of immunohistochemistry (IHC). Consequently we conducted a study utilising Biacore® analysis to investigate this relationship. Biacore® uses surface plasmon resonance to measure changes in the quantity of associated antibody-antigen to produce a real time kinetic measurement. We compared the affinity and IHC of several monoclonal antibodies including ER, PGR, CD3 and CD8 at equal concentrations. Here we present our data revealing the relationship between antibody affinity and IHC performance and review the role of other factors critical to antibody-antigen associations in FFPE tissue.

11.45 -­ 12.15 A high-throughput strategy for validation of antibodies towards unknown proteins
Dr Kenneth Wester Depts. of Surgical Sciences and Genetics & Pathology, Uppsala
University, Sweden
A high-throughput strategy for validation of antibodies towards unknown proteins.Mono-specific polyclonal antibodies are generated in a high through-put setting, using recombinant proteins for immunization of rabbits. Protein Epitope Signature Tags (PrESTs) are selected using bioinformatic data, available from the sequencing of the human genome. The antibodies are used for antibody-based tissue profiling with the aim to create an atlas of protein expression patterns in normal human tissues and the most common tumours. The talk is mainly focused on the strategy of validation of these antibodies, some of which are designed to recognize poorly characterized or unknown poteins.  Two antibodies, recognizing different epitopes, are produced for each gene/protein. The technical validation of antibodies is done by immunostaining of PrEST-arrays, representing 384 antigens per tested antibody and western blot on tissue/cell extracts from 5 different tissues. Antibodies approved in these tests are further evaluated by immunohistochemistry on tissue micro arrays (TMAs) representing 17 normal tissue types, 11 tumour types and 2 cell lines, in duplicate or triplicate, in all 70 spots. The staining patterns are compared to existing literature and bioinformatic data. Also, staining patterns from antibodies against the same protein are compared and scored regarding similarity. For each antibody, the final validation is based on a comprehensive evaluation of all obtained data.

12.15 -­ 12.45 The various faces of RNA in situ hybridisation
Dr Anja Kolb-Kokocinski, The Wellcome Trust Sanger Institute, Cambridge, UK
RNA in situ hybridisation is a popular tool to study gene expression. The visualisation of the location of a specific mRNA can help to shed light on the regulation and function of a gene. This approach might also help to identify disease genes.

Data from a broad range of different high-throughput projects are available for the scientific community, using a number of different protocols and technologies. I will explore and compare some of these alternatives using mainly studies on mouse development.

12.45 - ­ 13.00 Whole Slide Image Analysis in ImmunoHistochemistry
Mr Brian McClintock, Aperio Technologies, UK
With the ScanScope system, you can scan and analyse entire slides. Not only will you work more efficiently, but your final results will be better, because you are not limited to analysing only small representative portions of a specimen.

No more hunting around with a microscope, trying to find representative image fields to capture. This time consuming and labour intensive step is replaced with a fast, automated scan of the entire slide. You will have the added advantage of never having

to worry about whether you missed something. The entire digital slide is always available for immediate inspection, along with all analysis results.

Live demonstration of image analysis of Immunohistochemistry will be shown.

13.00 ­- 14.00 Lunch in the exhibition hall

14.00 -­ 14.15 Latest developments available from Launch Diagnostics from antigen retrieval to detection Brian Murphy, Launch Diagnostics, Kent, UK

BioGenex has achieved worldwide recognition in clinical diagnostics, life science and drug discovery as a result of 25 years of excellence in innovation covering reagents as well as highly advanced automated cell and tissue testing instruments.

BioGenex continues to be a premier customer service provider and is continually striving towards excellence and to provide a "Total Solution" for automated cell and tissue analysis. Automated systems from BioGenex streamline operations in cellular pathology and provide effective tools for the detection and diagnosis of cancer and infectious diseases. In recent years automated systems for drug discovery and development have been developed allowing high-throughput drug target characterization and histological profiling of genes and proteins. Furthermore, as inventors of antigen retrieval, a wide variety of antigen retrieval solutions for difficult to stain tissues is available.

Now in partnership with Launch Diagnostics for the UK and ROI a review of recently introduced BioGenex products and a look to the next stage of this developing partnership will highlight the latest developments available from Launch Diagnostics from antigen retrieval to detection.

14:15 - 14:45 The role of immunohistochemistry in drug discovery
Dr Chris Womack, AstraZeneca, Macclesfield, UK
Immunohistochemistry is one of a number of technologies used in drug discovery. Using cancer drug discovery as an example, the talk will illustrate how IHC fits into an overall strategy in the drug development trail. The relationship to other technologies and interdependencies in the “cancer toolkit” will be explored together with the challenges of introducing high throughput and automation in the process and the impact on decision-making.

14:45 ­- 15:15 Advanced Training in Diagnostic Immunohistochemistry
Mrs Barbara A Totty, Histopathology Manager, Addenbrooke's Hospital, Cambridge
Immunohistochemistry is essential to many diagnoses in histopathology directly affecting treatment and patient care. Highly trained biomedical scientists are needed to deliver this service and there is an opportunity for advancing the role of a biomedical scientist to become competent in interpreting these tests contributing to the histological report. The Institute of Biomedical Science (IBMS) and Royal College of Pathologists (RCPath) are working together to develop a two stage training and assessment programme for the Diploma of Expert Practice in Immunocytochemistry. The talk is illustrated with case studies involving a selection of prognostic and predictive markers showing the importance of good quality immunohistochemistry ensuring the best possible outcome from the patient’s perspective.

15:15 ­- 15:45 Afternoon break

15.45 -­ 16:15 Bright lights on dark background: developments in immunofluorescence
Dr Will Howat, Cancer Research UK, Cambridge Research Institute, UKThe technique of immunofluorescence has been a mainstay of the research community. In recent years, a number of developments have been made in fluorophore chemistry, such as Alexa dyes and Quantum Dots, that increase the flexibility and sensitivity of fluorescence. In addition, detection systems, such as automated fluorescence capture, have also opened up new possibilities for the ease of visualisation. These recent developments will be discussed.

16.15 -­ 16.30 Dako LINK TM: Let the Work Flow....
Mr Steve Jones, Dako UK Limited
Let us consider what the lab does at its core? It is gathering, processing and sending information. Usually that information is in the form of a glass slide, but it is still information. From our extensive experience of over 40 years in this industry, we fully understand how labs function and the demands placed on them to perform at a higher level, increasing productivity and controlling costs. We believe ALL of these requests can be addressed with our new product, Dako LINK™.

16:30 ­- 16.45 Chairman's summing up and close



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