Estrogen receptor (ER) expression is an important determinant of breast cancer behavior and is critical for response to endocrine therapies such as tamoxifen and aromatase inhibitors. In current practice, ER expression is determined by assay of biopsy material. In more advanced disease, tissue assay may present practical difficulties and be associated with significant sampling error. This and other considerations motivated the development of ER imaging agents for positron emission tomography (PET), of which the most successful has been (18)F-16alpha-17beta-fluoroestradiol (FES). In this review, we highlight aspects of ER biology and the importance of the ER in breast cancer therapy; review the structure and synthesis of FES; describe its kinetics and safety/dosimetry data; and highlight validation studies. Also discussed are early results in patients using FES-PET to localize ER-expressing tumors and associated data pointing toward its accuracy as a predictive assay for breast cancer endocrine therapy. Finally, early data for tumors and sites other than breast cancer are mentioned. Preliminary data strongly point toward potential clinical utility for FES-PET, motivating further validation and future clinical trials with prospective endpoints tested under appropriate regulatory oversight.