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A novel opto-chemical sensor instrumentation based on fiber optical micro-sensors for the simultaneous detection of pO2 and pCO2 in tissues is presented. The adopted sensing principle for both parameters is the measurement of luminescence lifetime via phase modulation fluorometry. Respect to currently used blood-gas analysers that require blood sampling and are associated with blood loss, this instrumentation allows on-line continuous measurements. Compared to electrochemical micro sensors, it is advantageous because of intrinsic higher sensitivity, no analyte consumption and less electromagnetic interferences. The results of the laboratory characterization and of tests in in vivo experiments of the proposed instrumentation are reported.