A system and method to measure blood oxygenation levels and total hemoglobin on individually selected blood vessels, to provide a representation of the subject condition and of tissue perfusion that may be used for diagnosing specific tissue conditions. Reflection spectra from individual blood vessels or a collection of vessels are measured by using wide-field imaging for selecting target vessels and a narrow-field confocal detection system to enable measuring local blood oxygenation and hemoglobin. Optical fibers may be used to illuminate the target vessel and to detect light diffusively reflected therefrom. The reflection spectra may be analyzed in a spectrometer to extract the ratio of the deoxy- to oxyhemoglobin and to determine their absolute concentration for computing total hemoglobin levels. An alternative implementation uses image processing on camera images of a blood vessel, generated at an isosbestic wavelength of the deoxy- and oxyhemoglobin, and optionally also at neighboring wavelengths.