A method for validating the start-to-end accuracy of a 3-D ultrasound (US)-based patient positioning system for radiotherapy is described. A radiosensitive polymer gel is used to record the actual dose delivered to a rigid phantom after being positioned using 3-D US guidance. Comparison of the delivered dose with the treatment plan allows accuracy of the entire radiotherapy treatment process, from simulation to 3-D US guidance, and finally delivery of radiation, to be evaluated. The 3-D US patient positioning system has a number of features for achieving high accuracy and reducing operator dependence. These include using tracked 3-D US scans of the target anatomy acquired using a dedicated 3-D ultrasound probe during both the simulation and treatment sessions, automatic 3-D US-to-US registration and use of infrared LED (IRED) markers of the optical position-sensing system for registering simulation computed tomography to US data. The mean target localization accuracy of this system was 2.5 mm for four target locations inside the phantom, compared with 1.6 mm obtained using the conventional patient positioning method of laser alignment. Because the phantom is rigid, this represents the best possible set-up accuracy of the system. Thus, these results suggest that 3-D US-based target localization is practically feasible and potentially capable of increasing the accuracy of patient positioning for radiotherapy in sites where day-to-day organ shifts are greater than 1 mm in magnitude.