The microfluidic biofuel cells have the potential to be improved for officially using in biomedical devices as a power source. It is noted that the basic design criterion of the power generating device such as glucose biofuel cell is a challenging task, especially in the presence of body fluids. In this study, a new glucose/O2 microfluidic biofuel cell based on printed circuit board and double-sided pressure sensitive adhesive tape was demonstrated. The electrodes of microfluidic biofuel cell were reduced graphene oxide‑gold nanoparticles-poly neutral red composite linked to Aspergillus niger glucose oxidase as a bioanode and Mytheliophthora thermophile laccase for biocathode. The open circuit voltage (OCV) and maximum power density of the microfluidic biofuel cell operating in physiological serum (pH 5.5) have obtained 0.2 V and 3.6 μW cm−2 at a flow rate of 50 μL min−1, respectively. The features of this rapid prototyping fabrication method, such as need not access to expensive clean room facility and affordability make it highly suitable for utilizing in resource poor environment.