A system for intraoperatively providing anatomical guidance in a diagnostic or therapeutic procedure, comprising: (a) a first light source configured to emit a beam of visible light to an area of interest of a living subject; (b) a second light source configured to emit a beam of near-infrared light to the area of interest; (c) a handheld probe optically coupled to the second light source but not optically coupled to the first light source, comprising an optical fiber configured to deliver the emitted beam of near-infrared light to illuminate the area of interest and configured to collect light that is scattered or emitted from a contrast agent (132a, 132b) introduced into target tissues in the area of interest, in response to illumination by the second light source; (d) a first imaging device optically coupled to the handheld probe and configured to detect the collected light (140a, 140b) and to generate a corresponding signal that comprises collected light data, and wherein the handheld probe is further configured to transmit the collected light (140a, 140b) to the first imaging device through the optical fiber; (e) a second imaging device configured to detect visible light that is emitted from the area of interest in response to illumination by the first light source and to generate a corresponding signal comprising visible light data; (f) a third imaging device configured to detect near-infrared light having a first predetermined wavelength that is emitted from the area of interest in response to illumination by the second light source and to generate a corresponding signal comprising a first set of near-infrared light data; (g) a fourth imaging device configured to detect near-infrared light having a second predetermined wavelength that is different from the first predetermined wavelength and that is emitted from the area of interest in response to illumination by the second light source, and to generate a corresponding signal comprising a second set of near-infrared light data; (h) a display for displaying at least one visual representation of data; and (i) a controller in communication with each of the first light source, second light source, first imaging device, second imaging device, third imaging device, fourth imaging device, and display, and programmed to generate at least one real-time integrated visual representation of the area of interest from each of the collected light data, visible light data, first set of near-infrared light data, and second set of near-infrared light data and to display the at least one realtime visual representation on the display, for guidance during the diagnostic or therapeutic procedure. (a) a first light source configured to emit a beam of visible light to an area of interest of a living subject; (b) a second light source configured to emit a beam of near-infrared light to the area of interest; (c) a handheld probe optically coupled to the second light source but not optically coupled to the first light source, comprising an optical fiber configured to deliver the emitted beam of near-infrared light to illuminate the area of interest and configured to collect light that is scattered or emitted from a contrast agent (132a, 132b) introduced into target tissues in the area of interest, in response to illumination by the second light source; (d) a first imaging device optically coupled to the handheld probe and configured to detect the collected light (140a, 140b) and to generate a corresponding signal that comprises collected light data, and wherein the handheld probe is further configured to transmit the collected light (140a, 140b) to the first imaging device through the optical fiber; (e) a second imaging device configured to detect visible light that is emitted from the area of interest in response to illumination by the first light source and to generate a corresponding signal comprising visible light data; (f) a third imaging device configured to detect near-infrared light having a first predetermined wavelength that is emitted from the area of interest in response to illumination by the second light source and to generate a corresponding signal comprising a first set of near-infrared light data; (g) a fourth imaging device configured to detect near-infrared light having a second predetermined wavelength that is different from the first predetermined wavelength and that is emitted from the area of interest in response to illumination by the second light source, and to generate a corresponding signal comprising a second set of near-infrared light data; (h) a display for displaying at least one visual representation of data; and (i) a controller in communication with each of the first light source, second light source, first imaging device, second imaging device, third imaging device, fourth imaging device, and display, and programmed to generate at least one real-time integrated visual representation of the area of interest from each of the collected light data, visible light data, first set of near-infrared light data, and second set of near-infrared light data and to display the at least one realtime visual representation on the display, for guidance during the diagnostic or therapeutic procedure.