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Continuous intra-arterial blood pH monitoring is highly desirable in clinical practice. However, devices with appreciable accuracy are still not commercially available to date. In this study, we present a fiber-optic fluorosensor that can be used to continuously and accurately measure blood pH changes. The pH sensor is developed based on a proton-sensitive fluorescence dye, N-allyl-4-(4'-methyl-piperazinyl)-1,8-naphthalimide, which is bonded covalently to an optical fiber through heat polymerization. Fluorescence intensity was recorded after the sensor was exposed to different pH buffer solutions or intra-arterial blood in rabbits. Fluorescence intensity with emission peak at 510 nm decreased immediately as the blood pH increased. Linear and reproducible responses were observed when pH ranges from 6.8 to 8.0 with res olution of 0.03 pH units. The correlation coefficient between the pH sensor and the conventional blood gas analyzer was 0.93 in vivo (n = 75, p <; 0.001) with a bias and precision of -0.02 ± 0.08 pH units. The pH sensor was stable during measurement for at least 72 h. The pH sensor is not sensitive to fluctuations of vari ous ions' concentrations and plasma osmosis at pathophysiological limits, suggesting that it is useful for the continuous measurement of blood pH at various clinical settings.