Protein expression analysis using quantitative fluorescence image analysis on tissue microarray slides

Biotechniques. 2002 Apr;32(4):924-6, 928-30, 932. doi: 10.2144/02324pt04.

Abstract

We have developed a tissue microarray (TMA)-based quantitative fluorescence image analysis (QFIA) method in which protein markers on TMA sections were labeled by immunofluorescence using tyramide signal amplification and a quantitative fluorescence detection system. Using this method, BRCA1 protein expression patterns were studied in the TMA sections of cell lines with known levels of BRCA1 expression and in a small group of human tissue samples obtained from sporadic epithelial ovarian cancers, their corresponding adjacent dysplastic fields, and distant non-tumor areas. We detected distinctive BRCA1 expression patterns in high-grade and low-grade sporadic epithelial ovarian cancers and their associated adjacent dysplastic fields. However, such patterns of expression could not be adequately detected by traditional immunohistochemical staining methods. TMA-QFIA provides a sensitive, automated, and quantitative measurement of protein expression on archived tissue and cell samples and will be a useful tool for protein-level molecular profiling analyses.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • BRCA1 Protein / analysis*
  • Blotting, Western
  • Breast Neoplasms / chemistry
  • Female
  • Fluorescent Antibody Technique / methods*
  • Histocytological Preparation Techniques / methods*
  • Humans
  • Immunohistochemistry
  • Ovarian Neoplasms / chemistry
  • Tumor Cells, Cultured

Substances

  • BRCA1 Protein