3rd Improving Immunohistochemistry Discussion Forum-17th Oct 2012

3rd Improving Immunohistochemistry Discussion Forum

The Stevenage Bioscience Catalyst , Gunnels Wood Road, Stevenage, UK: Wednesday, 17 October 2012

This discussion forum focuses on the technique of immunohistochemistry, from DAB to double staining. The invited experts will give short overview presentations on their sub-topics, with the majority of the day dedicated to informal question and answer troubleshooting sessions.

Meeting Chair: Dr Will Howat, Head of Histopathology/ISH at Cambridge Research Institute, Cancer Research UK

This event has CPD accreditation

On registration please submit your questions to the panel that will be asked by the chair on the day of the event

9:00 – 9:30 Registration

9:30 – 9:35 Introduction by Meeting Coordinator: Dr Astrid Englezou , Euroscicon, London, UK

9:35 – 9:40 Introduction by the Chair: Dr Will Howat, Head of Histopathology/ISH at Cambridge Research Institute, Cancer Research UK

9:40– 10:00 Title to be confirmed
Dr Will Howat, Head of Histopathology/ISH at Cambridge Research Institute, Cancer Research UK

10:00 – 10:20 Immunohistochemistry for Glycol Methacrylate embedded samples
Dr Susan J Wilson PhD, Head of the Histochemistry Research Unit, Southampton General Hospital
Glycol methacrylate (GMA) was introduced in the 1960s and is a hydrophilic acrylic resin. By using a modified processing and embedding procedure it can be used for immunohistochemical studies. This offers several advantages over traditional techniques including good antigen preservation, excellent morphology and antigen localisation, no need for antigen retrieval, cutting of numerous serial sections is possible and co-localisation studies. We use an avidin-biotin peroxidase detection system and have applied this methodology to a wide range of samples including lung, gut, kidney, skin, conjunctiva, tissue culture monolayers and dispersed cells. Details of the technique including fixation, processing, sectioning and the immunohistochemical protocol will be discussed.

10:20 – 10:40 Talk to be confirmed
Arthur Lewis, MedImmune, UK

10:40 – 11:00 Immunohistochemistry in The Human Protein Atlas project
Urban Ryberg, The Human Protein Atlas project, Uppsala, Sweden

In a large scale high throughput project, aiming to produce antibodies towards all non-redundant human proteins, it is necessary to use standardised protocols. The large numbers of antibodies to be tested makes it impossible to optimise protocols for each antibody and therefore we have tried to find an immunohistochemistry protocol that works for most of our antibodies and that makes it possible to reproduce results.

11:00 – 11:20 Mid-morning Break

Please try to visit all the exhibition stands during your day at this event. Not only do our sponsors enable Euroscicon to keep the registration fees competitive, but they are also here specifically to talk to you

11:20 – 11:40 Making the most of tissue microarrays
Dr Anthony Warford, University of Westminster, London, UK
Tissue microarrays (TMAs) have revolutionised the way in which research is done in cellular pathology. In essence, they miniaturise the amount of tissue and time required to generate statistically meaningful results. To achieve this, TMAs must be prepared carefully. First and foremost, tissue cores from each donor sample must be representative of the histology or pathology. To achieve this guided tissue selection must be used and consideration given to the diameter and number of cores removed from each donor sample. Led by the research question to be answered the number of donor samples in each TMA should also be carefully considered. Properly made hundreds of sections can be cut from a TMA block and these should be protected from antigen degradation before staining. Lastly, once staining has been completed, analysis should be undertaken in a consistent manner to maximise the return on the investment in TMA preparation.

11:40 – 12:00 Quantitation of protein biomarker concentrations with sub-cellular resolution from routine clinical FFPE tissues

Dr Julian Beesley, HistoRx Europe, USA

HistoRx, www.historx.com is developing unique diagnostic solutions to unlock new approaches to individualised patient care.

HistoRx’s proprietary AQUA® technology uses quantitative IHC to deliver objective and reproducible multiparametric analysis of proteins in routine clinical specimens, providing new insights into cancer treatment. The Company is applying this expertise to build a proprietary pipeline of diagnostic candidates and to help biopharmaceutical customers advance their drug development programmes.

12:00 – 12:40 Working Lunch

Please collect your lunch and take it to your discussion table (Session 1)

This is also a good time to fill out your feedback forms

12:40 – 14:10 Discussion Group Sessions 1 - 3

• Round table discussion groups (20 minutes each) will be held throughout the afternoon
• Delegates will rotate so that they may participate in all the discussion tables
• All delegates will also be allocated a session for visiting the exhibition stands
• Where appropriate delegates will be able to bring their samples to the discussions
• See end of agenda for description of discussion tables

14:10 – 14:30 Tissue Cross-reactivity Studies: current considerations
Joanne Mitchell, Huntingdon Life Sciences, Huntingdon, UK

An overview of tissue cross-reactivity studies will be presented, this year focusing on current methodologies and workflows, latest changes to guidelines and a review of published survey results on the use of the tissue cross-reactivity assay in regulatory studies.

14:30 – 15:00 Talk to be confirmed

Advanced Cell Diagnostics, Inc, USA

15:00 – 16:20 Discussion Group Sessions 4 - 6

16:20 – 17:00 Question and Answer Session

This session will include summing up of the discussion tables and questions submitted both prior to the meeting and throughout the day

17:00 Chairman’s Summing Up and Feedback Prize Draw

Discussion tables

Table A: Will Howat

Table B: Susan J Wilson

Table C: Urban Ryberg

Table D: Anthony Warford,

Table E: Julian Beesley

Table F: Arthur Lewis

Keywords: Tissue Microarrays, immunohistochemistry, fixation, imaging, quantitation,glycol methacrylate resin, immunohistochemistry, fixation, Immunohistochemistry, tissue micro array, formalin fixed paraffin embedded material, high through put, quantifying biomarkers, clinical tissues, automated-analysis

About the Chair

Will Howat graduated with a BSC (Hons) in Immunology & Pharmacology from the University of Strathclyde, before gaining a PhD in Pathology from the University of Southampton. After two post-doctoral positions in Southampton, he moved to the Wellcome Trust Sanger Institute in Cambridge as the leader of Research & Development for the Immunohistochemistry group of the Atlas of Protein Expression project. He is now with Cancer Research UK as the head the Histopathology/ISH facility at the Cambridge Research Institute.

About the Speakers
Anthony (Tony) Warford expertise is in molecular histopathology. He has set up and managed laboratories in the UK health service, academic institutions, biotechnology and Pharmaceutical companies. Technology developments he has spearheaded include the introduction of diagnostic immunohistochemical methods, validation of antibodies for use as biomarkers, production of probes and methods for in situ hybridisation and supervision and interpretation of GLP tissue based safety studies of potential therapeutic antibodies. Concurrently he has championed quality assurance programmes in histopathology and automation of immunohistochemistry coupled with image capture and analysis. He has also run laboratory safety and human bio-banking programmes. He has published in these fields and shared experience with fellow scientists by organising wet workshops, chairing symposia and lecturing in many countries.

Susan Wilson initially trained as a NHS Biomedical Scientist in histopathology. After gaining her Fellowship of the Institute of Biomedical Sciences in Cellular Pathology, she took a position at the University of Southampton. Susan was a founding member of Professor Holgate’s asthma bronchoscopy research programme, providing histopathology support which furthered the understanding of the pathobiology of asthma and during this time she studied for a PhD in Mucosal Inflammation. Susan has over 25 years experience in histopathology and now heads the Histochemistry Research Unit (HRU) within the Faculty of Medicine. The HRU is a core facility offering a full histopathology support service. The Unit is internationally recognised for its histopathology expertise, being particularly known for the use of glycol methacrylate (GMA) resin embedding for immunohistochemistry procedures.

Joanne Mitchell holds a B.Sc. Hons degree in Applied Biology with European Studies and a Masters degree in Biomedical Science. She has over 13 years experience in immunohistochemistry (IHC) and histopathology gained from five years working at Pharmagene in ‘Molecular Imaging’ pre-clinical research projects, three years managing operations in ‘The Atlas Project’ IHC laboratory at The Sanger Institute and two years setting-up and running a Histopathology/IHC Core Facility for Cancer Research UK. Jo joined HLS in 2008 as Head of Special Pathology Services. She is responsibile for IHC, tissue cross-reactivity, electron microscopy and micro-autoradiography studies, working to GLP.

Julian Beesley has over 22 years’ experience as a research scientist and senior manager in discovery research within the pharmaceutical sector and over 12 years’ involvement in developing biotech businesses, operating widely throughout Europe, the USA, Asia and Japan.