Optical monitoring of various tissue physiological and biochemical parameters in real-time represents a significant new approach and a tool for better clinical diagnosis. The Tissue Spectroscope (TiSpec), developed and applied in experimental and clinical situations, is the first medical device that enables the real-time monitoring of three parameters representing the vitality of the tissue. Tissue vitality, which is correlated to the oxygen balance in the tissue, is defined as the ratio between O2 supply and O2 demand. The TiSpec enables the monitoring of microcirculatory blood flow (O2 supply), mitochondrial NADH redox state (O2 balance), and tissue reflectance, which correlates to blood volume. We describe in detail the theoretical basis for the monitoring of the three parameters and the technological aspects of the TiSpec. The comparison between the TiSpec and the existing single parameter monitoring instruments shows a statistically significant correlation as evaluated in vitro as well as in various in vivo animal models. The results presented originated in a pilot study performed in vivo in experimental animals. Further research is needed to apply this technology clinically. The clinical applications of the TiSpec include two situations where the knowledge of tissue vitality can improve clinical practice. The major application is the monitoring of "nonvital" organs of the body [i.e., the skin, gastrointestinal (G-I) tract, urethra] in emergency situations, such as in the operating rooms and intensive care units. Also, the monitoring of specific (vital) organs, such as the brain or the heart, during surgical procedure is of practical importance.