Continuous measurement of dissolved oxygen (DO) is essential for water quality monitoring and biomedical applications. Here, a phosphorescence quenching-based intelligent dissolved oxygen sensor on an optofluidic platform for continuous measurement of dissolved oxygen is presented. A high sensitivity dissolved oxygen-sensing membrane was prepared by coating the phosphorescence indicator of platinum(II) meso-tetrakis(pentafluorophenyl)porphyrin (PtTFPP) on the surface of the microfluidic channels composed of polydimethylsiloxane (PDMS) microstructure arrays. Then, oxygen could be determined by its quenching effect on the phosphorescence, according to Stern–Volmer model. The intelligent sensor abandons complicated optical or electrical design and uses a photomultiplier (PMT) counter in cooperation with a mobile phone application program to measure phosphorescence intensity, so as to realize continuous, intelligent and real-time dissolved oxygen analysis. Owing to the combination of the microfluidic-based highly sensitive oxygen sensing membrane with a reliable phosphorescent intensity detection module, the intelligent sensor achieves a low limit of detection (LOD) of 0.01 mg/L, a high sensitivity of 16.9 and a short response time (22 s). Different natural water samples were successfully analyzed using the intelligent sensor, and results demonstrated that the sensor features a high accuracy. The sensor combines the oxygen sensing mechanism with optofluidics and electronics, providing a miniaturized and intelligent detection platform for practical oxygen analysis in different application fields.