OBJECTIVE Prostate movement is a major consideration in the formation of target volumes for conformal radiation therapy of prostate cancer. The goal of this study was to determine the technical feasibility of using implanted radiopaque markers and digital imaging to localize the prostate at the time of treatment, thus allowing for reduction of the margin required for uncertainty in target position. METHODS Radiopaque markers implanted around the prostate prior to treatment are visible on electronic radiographs generated with a portal imager or diagnostic imaging device. The locations of the images of these markers on the digital radiographs were automatically determined by a template-matching algorithm. The coordinates of the markers were found by projecting rays through the marker locations on orthogonal radiographs using a three-dimensional (3D) point-matching algorithm. Prostate and/or patient movement was inferred from the marker displacements. Images generated from known movements of a phantom with implanted markers were tested with this algorithm. Locations of markers from daily images of patients with implanted markers were determined by both manual and automatic techniques to determine the efficacy of automated localization on typical clinical images. RESULTS Prostate movements can be automatically detected in a phantom using low-energy photons within 30 s after image acquisition and with a precision of better than 1 mm in translation and 1 degree in rotation (indistinguishable from the uncertainty in measuring precision). CONCLUSIONS The studies show that on-line repositioning of the patient based on localization of the markers at the time of treatment is feasible, and may reduce the uncertainty in prostate location when combined with practical on-line repositioning techniques.