Nanocellulose films, made from biodegradable nanoscale cellulose fibers, exhibit excellent mechanical strength, low thermal expansion, high transparency, and outstanding environmental attributes. Microfluidic biosensors, known for their high integration, automation, sensitivity, and rapid response times, are widely used in environmental monitoring, clinical diagnostics, and food safety testing. Nanocellulose films are particularly suitable for fabricating microfluidic biosensors due to their superior barrier properties, liquid-carrying capacity, and plasticity, significantly enhancing the repeatability and selectivity of biological detection while reducing waste. In this study, we developed a silver electrode electrochemical sensor with high stability and repeatability using inkjet printing technology for the first time, successfully applied in glucose testing. The fabrication time is less than 15 min. Notably, we explored the impact of particle size and distribution uniformity of silver electrodes on their conductivity and stability. More importantly, based on the redox reaction of silver nitrate and sodium borohydride, we established a production process with a high yield of electrodes. We hope this research advances the application of nanocellulose film-based electrochemical sensors in biological detection.