Visible light and near infrared light interact with biological tissue by absorption and scattering. Diffuse optical imaging and spectroscopy reconstructs tissue physiologic parameters based on noninvasive measurement of tissue optical properties. This technology can be used to differentiate physiologic and molecular signatures of both malignant and benign tissues, as they relate to the area of cancer research. Major advantages are the use of non-ionizing radiation, real-time continuous data acquisition, low cost, and portability. Limitations include low spatial resolution and limited reproducibility. This paper reviews the currently available state-of-the-art technologies for diffuse optical imaging and spectroscopy and their applications in cancer research.