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Related Experiment Videos

Expression profiling by high-throughput immunohistochemistry.

Anthony Warford1, William Howat, John McCafferty

  • 1Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridgeshire CB10 1SA, UK.

Journal of Immunological Methods
|July 21, 2004
PubMed
Summary

High-throughput immunohistochemistry (IHC) is becoming feasible due to advances in tissue microarrays, automation, and image analysis. Successful application requires careful consideration of antibody quality and validation for reliable protein expression profiling.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Pathology

Background:

  • Immunohistochemistry (IHC) is a crucial technique for visualizing protein expression in tissues at cellular and subcellular levels.
  • Recent technological advancements are enabling IHC to be utilized as a high-throughput expression profiling system.

Purpose of the Study:

  • To review recent developments in immunohistochemistry (IHC) that support its use as a high-throughput expression profiling system.
  • To consider the key technologies and antibody-related issues impacting the success of high-throughput IHC.

Main Methods:

  • Review of advancements in tissue microarray (TMA) creation and utilization.
  • Evaluation of automated IHC systems and their role in high-throughput applications.
  • Analysis of progress in image capture and analysis techniques for IHC data.

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  • Consideration of antibody generation, screening, specificity, sensitivity, and fixation choices.

    • Main Results:

    • Tissue microarrays, automated IHC, and advanced image analysis are key enabling technologies for high-throughput IHC.
    • The success of high-throughput IHC is significantly influenced by the quality and validation of antibodies used.

    Conclusions:

    • High-throughput IHC is a promising approach for large-scale protein expression profiling.
    • Addressing technological and antibody-related challenges is essential for the widespread adoption and reliability of high-throughput IHC.