Time-resolved spectroscopy and near infrared imaging enhanced by receptor-targeted contrast agents for prostate cancer detection will be presented. Two contrast agents, Cybesin and Cytate, were investigated using time-resolved spectroscopy in aqueous solution and cancerous and normal prostate tissues. The time evolution of the fluorescent dipole in solution was studied using a system of first-order linear differential equations containing two main parameters: the decay rate of emission and the rate of one orthogonal emission component transferring to another. An analytical polarization model was developed and used to extract rotational times and fluorescence anisotropies of the contrast agents in prostate tissues. The differences of rotational times and polarization anisotropies were observed for Cybesin (Cytate) in cancerous and normal prostate tissue, which reflect preferred bond of contrast agents and cancerous tissue cells. The conjugation of Cybesin (Cytate) to prostate cancerous cells offers high contrast between normal and cancerous tissues.