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Automated Analysis of Tissue Microarrays

  • Marisa Dolled-FilhartEmail author
  • Mark Gustavson
  • Robert L. Camp
  • David L. Rimm
  • John L. Tonkinson
  • Jason Christiansen
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 664)

Abstract

The analysis of protein expression in tissue by immunohistochemistry (IHC) presents three significant challenges. They are (1) the time-consuming nature of pathologist-based scoring of slides; (2) the need for objective quantification and localization of protein expression; and (3) the need for a highly reproducible measurement to limit intra- and inter-observer variability. While there are a variety of commercially available platforms for automated chromagen-based and fluorescence-based image acquisition of tissue microarrays, this chapter is focused on the analysis of fluorescent images by AQUA® analysis (Automated QUantitative Analysis) and the solutions offered by such a method for research and diagnostics. AQUA analysis is a method for molecularly defining regions of interest or “compartments” within a tissue section. The methodology can be utilized with tissue microarrays to provide rapid, quantitative, localized, and reproducible protein expression data that can then be used to identify statistically relevant correlations in populations. Ultimately this allows for a multiplexed, objective and standardized quantitative approach for biomarker research and diagnostic assay development for protein expression in tissue.

Key words

Immunohistochemistry Automated analysis AQUA Tissue microarrays Quantitative analysis Biomarkers Immunofluorescence 

Notes

Acknowledgments

RLC is supported by grants from the NIH/NCI including R21 CA 125277 and R21 CA 116265. DLR is supported by grants from the NIH including RO-1 CA 114277, R33 CA 106709 and R33 CA 110511.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Marisa Dolled-Filhart
    • 1
    Email author
  • Mark Gustavson
    • 1
  • Robert L. Camp
    • 2
  • David L. Rimm
    • 2
  • John L. Tonkinson
    • 1
  • Jason Christiansen
    • 1
  1. 1.HistoRx, IncNew HavenUSA
  2. 2.Department of PathologyYale UniversityNew HavenUSA

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