This study presents a rapid, sensitive and selective sensor based on electrospun nanofibers on laser-patterned electrodes, in which the laser micromachining process can directly fabricate the graphene electrode device for the electronical detection of glucose molecule. The layer of graphene film was formed on the glass substrate by a screen printing technique, and then the graphene electrodes can be fabricated by the ultraviolet (UV) nanosecond laser process with the wavelength of 355 nm. Based on the controlled the laser fluence and pulses, the electrode gap of 60 μm within the device can be fabricated. The polyvinyl alcohol (PVA) and glucose oxidase (GOD) composite nanofiber with weight percentage of 8% can be used by the electrospun process with used Glutaraldehyde(GA) steam for cross linking process. After that, it is blended with the conductivity of poly(3,4-ethylenedioxythiophene): poly (styrene sulfonate) (PEDOT:PSS), and the foldable nanofiber structures was made by electrospun process on the graphene electrode device. Finally, the electrical detection of graphene electrode device with the nanofibers at different concentrations of glucose can be measured, resulting in the linear variation is detecting the glucose concentration ranging form 0.01 to 3.12 mM. This work indicated that the low concentration and highly sensitive can be used for electronic biosensors
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