Automated evaluation and normalization of immunohistochemistry on tissue microarrays with a DNA microarray scanner

Biotechniques. 2003 Jul;35(1):164-8. doi: 10.2144/03351md04.

Abstract

Hundreds of tissue samples may be assembled in a tissue microarray format for simultaneous immunostaining assessment of protein expression profiling. A DNA microarray two-color laser scanner was used for automated analysis of tissue microarray indirect immunofluorescence. On sections from both a human lung adenocarcinoma and a squamous cell carcinoma tissue microarray, fluorescence intensity for two epidermal growth factor receptors (EGFR and c-erbB2) correlates with diagnostic pathologic assessment, indicating that immunohistochemistry quantitation can be achieved. Importantly, double-label indirect immunofluorescence detection with the cDNA scanner demonstrates that one reference antigen can normalize tumor marker immunosignal for the cellular content of tissue microarray tissue cores. Therefore, DNA microarray scanners and associated image analysis software provide general and efficient analysis of tissue microarray immunostaining, including estimation of specific protein expression levels.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adenocarcinoma / genetics
  • Adenocarcinoma / metabolism*
  • Carcinoma, Squamous Cell / genetics
  • Carcinoma, Squamous Cell / metabolism*
  • ErbB Receptors / analysis
  • ErbB Receptors / genetics
  • ErbB Receptors / metabolism*
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation, Neoplastic / physiology
  • Humans
  • Immunohistochemistry / methods*
  • Lung Neoplasms / genetics
  • Lung Neoplasms / metabolism
  • Oligonucleotide Array Sequence Analysis / methods*
  • Quality Control
  • Receptor, ErbB-2 / analysis
  • Receptor, ErbB-2 / genetics
  • Receptor, ErbB-2 / metabolism*
  • Robotics / methods
  • Subtraction Technique
  • Tumor Cells, Cultured

Substances

  • ErbB Receptors
  • Receptor, ErbB-2