Improving Immunohistochemistry

London
Friday, 07 April 2006

Improving Immunohistochemistry
Friday, 07 April 2006 09:15 - 16:30

Birkbeck College
Room B04
Malet Street
London
WC1E 7HX
United Kingdom

Map and Directions

This years meeting will focus on gene expression.  Presentations will consider the fixation of nucleic acids, compare their demonstration with proteins and explore the role of fluorescence detection methods.  For the first time, presentations will be given on both immunohistochemistry based protein atlas projects. Dr Tony Warford, Wellcome Trust Sanger Institute, Cambridge (meetings chair)


 

09:15 – 09:45              Registration – Tea, coffee and biscuits

 

09:45 - 10:00               Introduction by the Chair: Dr Tony Warford - Wellcome Trust Sanger Institute

 

10:00 - 10:30              Demonstrating nucleic acids in paraffin wax tissue - Dr Tony Warford
The interaction of fixatives with nucleic acids has not been studied to the same extent as with proteins.  With the demonstration of nucleic acids either in situ or after extraction being increasingly used it is important to understand these.  In this presentation interactions will be reviewed for commonly used fixatives and fixative mixtures from chemical and practical viewpoints.  Particular emphasis will be placed on the effect of fixation on subsequent in situ hybridisation and PCR amplification analysis methods.


10:30 – 11:00              The correlation of mRNA and protein expression in the morphological and non - morphological analysis of human tissues -  Dr Keith Page, Pharmagene plc

The combined use of genomic and morphological (mRNA and protein visualisation and cellular phenotyping) techniques in human tissues, in the discovery and validation of therapeutic targets for cystic fibrosis.

 

11:00 – 11:30              Morning coffee/tea break in the exhibition hall

 

11:30 – 11:45               Isoform Specific Antibodies in Immunohistochemistry - Dr Ian Milton - Novocastra Laboratories

Immunohistochemistry has been a routine for over 20 years. Traditionally antibodies used in IHC have been selected on the basis of their visual performance. The sequencing of the human genome and associated research has revolutionised our understanding of different isoforms of genes and proteins. This information now allows us to devise a new generation of isoform specific antibodies which can be used to provide added clinical information.

11:45 – 12.15              Developing an Atlas of protein expression, using recombinant antibodies and tissue microarrays - Dr John  McCafferty - Wellcome Trust Sanger Institute
The "Atlas of Gene Expression" project is using IHC to describe protein expression in tissues, using recombinant antibodies as probes. Developing this into an efficient high throughput process has required optimisation and integration of a range of diverse activities including antigen production and antibody selection by phage display. Recent developments now make it possible to contemplate using IHC as a high throughput expression profiling system, (including the use of tissue microarrays, automated IHC and image capture/analysis). The presentation will describe our work in improving and incorporating all of these activities into a process for efficient generation of protein expression data.


12.15 - 12.30           The use of on-slide controls for the scoring of Her2/neu IHC slides, using the Ariol system - Mr Rob Sykes, Applied Imaging

Currenty IHC is becoming more and more standardised, with staining protocols and antigen retrieval being automated very successfully. With the advent of therapy linked IHC tests that require quantification of end points, the use of imaging systems may provide a more robust and sensitive way of assessing these. 


12.30 – 12.45              OmniMap – Ventana’s biotin free detection system - Dr Andre Carvalho, MDS Ventana Medical Systems
Biotin free detection system for IHC.  Brief comparison and advantages over Ventana's DABMap kit.

12:45 - 13:45               Lunch in the exhibition area

 

13.45 – 14.00              Tumor biology is King (Queen): Combining validated IHC and FISH assays to optimize and individualize breast cancer treatmentDr Christof Henne, DakoCytomation Ltd, Germany

Focus on ER, PR, HER2, and TOP2A (Topo IIalpha) used as predictive (and prognostic) tumor markers to select optimal treatment for breast cancer patients. The need for assay standardization and validation. How current molecular methodologies can be used to subdivide breast cancer in clinically meaningful subtypes with different treatment needs.


14.00 – 14.30        Improving Immunohistochemistry -  Prof I Ellis, - University of Nottingham

Identification of protein expression using immunocytochemistry has become standard technology in human tissue diagnosis and diagnostic and research evaluation of biomarkers.  Future challenges involve developing methodologies to ensure standardisation and reproducibility of staining methodology and interpretation.  Improvements in automation should lead to improved methodology standardisation and development of sophisticated image analysis techniques applicable to high resolution digitised images can provide solutions to reproducible and semi-automated interpretative analysis.  Improved methodologies to allow multiplexing through use of nano particle technology are likely to impact on research applications in immunocytochemistry.

  

14.30 – 15.00              Antibody-Based Proteomics For Human Tissue Profiling - Dr. Fredrik Ponten – Human Proteome Resource, Sweden

A multi-disciplinary research program to create a “Human Proteome Resource” was started in 2003. This program includes systematic exploration of the human proteome using antibody-based tissue proteomics, combining high-throughput generation of mono-specific antibodies (msAb) with protein profiling in human tissues and cells using tissue microarrays. Recombinant protein fragments selected from unique regions called Protein Epitope Signatures Tags (PrESTs) were used as imunogens to generate msAbs. Tissue microarrays including >700 spots of normal and cancer tissues was used for an automated array-based high throughput technique for expression profiling.  One important objective was to generate a protein atlas, as a knowledge-base with regard to the structural and temporal expression of proteins in various cells and tissues (www.proteinatlas.org).


15.00 – 15.15              Integrating Digital Pathology  - Mr Brian McClintock, Aperio Technologies
Now that Virtual Microscopy is a reality, there is a need to integrate the images with Gross images, Radiology and other patient data, to achieve an integrated electronic patient record.

15:15 – 15.45              Afternoon Tea/Coffee and cakes in the exhibition area

15:45 – 16:15            Quantum dot based in-situ gene expression profiling - Dr Richard Byers - Senior Lecturer in Pathology, University of Manchester and Consultant Pathologist, Manchester Royal Infirmary

Quantum dots have recently been used for bioimaging by immunofluorescence, and for molecule and cell labelling. They are fluorescent semiconductor nanocrystals possessing the unique properties of extremely high fluorescence efficiency, lack of photobleaching and long fluorescence lifetime, making them near-optimal for many fluorescent applications.  In addition, whilst their excitation wavelength is constant, their emission wavelength is sharp, symmetrical and tunable dependent upon diameter, with potential for multicolor staining. We have used QD-labeling of DNA oligonucleotide probes and spectral imaging to produce a novel technique for multiplex immunoflourescence and in-situ hybridisation.


16.15 - 16.30              GenePaintTM system for advanced in situ studies (ISH, FISH, IHC)

Dr Christoph Jung, Tecan Switzerland AG

One prerequisite for large scale projects such as atlas of gene expression is a standardized and automated workflow for in situ studies. The GenePaintTM system--invented by Gregor Eichele of the Max Planck Institute of Experimental Endocrinology, Hannover, Germany-- automates all of the steps required for performing ISH and IHC of tissue sections on microscope slide formats. It helps to deliver consistent and reproducible staining results of high quality and it saves on the amounts of reagents used.

 

 



16:30 - 16:45                Discussion and Close


 

Registraton fees

  • Standard fee - £440
  • Academic fee - £220
  • Student fee- £140
  • IBMS members fee - £199

 

 

Contact Details

Payment Instructions

    • Payment can be made by Visa or Mastercard online by secure server or by post
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    Sally Wheatland
    Euroscicon
    PO Box 49717
    London
    N20 8WH.

    Payment must be received prior to the meeting

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