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In this paper, an arrayed glucose biosensor is integrated to the poly-dimethylsiloxane microchannel as the microfluidic device for measuring the characteristics of the arrayed glucose biosensor at dynamical conditions. The radio frequency sputtering system and screen printed technology are used to fabricate the arrayed glucose biosensor. Furthermore, the computer numerical control technique is used to fabricate the microchannel mold. The polypyrrole differential reference electrode has good stable characteristics and provides a stable potential for the arrayed pH sensor. At static conditions, the average sensitivity and linearity of the arrayed pH sensor are 57.85 mV/pH and 0.977, respectively. And at dynamical conditions with 20 mL/min flow rate, the average sensitivity and linearity of the arrayed pH sensor are 66.73 mV/pH and 0.993, respectively. The arrayed pH sensor is dropped glucose sensing membranes as an arrayed glucose biosensor. The arrayed glucose biosensor is measured in different glucose concentrations at static conditions, and the average sensitivity and linearity of the arrayed glucose biosensor are 11.84 mV(100 mg/dl)-1 and 0.995, respectively. At dynamical conditions with 25 mL/min flow rate, the average sensitivity and linearity of the arrayed glucose biosensor are 30.41 mV(100 mg/dl)-1 and 0.975, respectively.