Optical tomography with ultrasound (US) localization uses coregistered ultrasound images to guide optical imaging reconstruction. To simultaneously acquire US images and optical measurements, the authors used a hand-held probe consisting of a commercial US transducer and near-infrared optical imaging sensors of multiple wavelengths. A novel image scheme was used to map the ultrasound-visible lesions for optical imaging reconstruction. As a result, the problem of intense light scattering caused by breast tissue was overcome and reliable tumor hemoglobin concentration and blood oxygen saturation distributions from a group of patients were obtained. These functional parameters are valuable for aiding US diagnosis and for assessing chemotherapy response.