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This paper presents a transrectal dual-modality near-infrared (NIR) diffuse optical tomography technique coupled with ultrasonography that provides an integrated method for detecting prostate cancer (PCa). The study that provides an alternative transrectal prostate imaging system stems from the perceived inadequacy of conventional transrectal ultrasound (TRUS) in PCa imaging. The transrectally applied diffuse optical tomography aims to characterize the spatially resolved optical properties of the intact prostate that are known to have intraorgan and intersubject heterogeneities. A canine model of PCa using canine transmissible venereal tumor was used to demonstrate the utility of this technology in detecting PCa. Tumors in the pelvic canal, including tumors in the prostate, were found to be detectable at 2-week postinjection based upon the NIR absorption contrast, which was detected a few weeks earlier than using NIR-reduced scattering and effective attenuation contrasts, as well as TRUS. Transrectal optical tomography detection of cancerous tissues in vivo in intact canine prostate based upon NIR contrasts may prove useful for diagnostic imaging of PCa and potentially aid in pretreatment planning for phototherapy applications.