Protein expression analysis using quantitative fluorescence image analysis on tissue microarray slides.
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.
Rao
JianYu
J
Department of Pathology and Laboratory Medicine, UCLA, Los Angeles, CA 90095, USA. jrao@mednet.ucla.edu
Seligson
David
D
Hemstreet
George P
GP
D019313:BRCA1 Protein / Q000032:analysis*; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000737:chemistry; D005260:Female; D005455:Fluorescent Antibody Technique / Q000379:methods*; D016591:Histocytological Preparation Techniques / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry; D010051:Ovarian Neoplasms / Q000737:chemistry; D014407:Tumor Cells, Cultured
D019313:BRCA1 Protein