Biomarkers based sensing in the realm of disease prevention, its diagnosis, and drug development has shown its unique importance in health care. In particular, the platelet level in blood is strongly linked with the development of several kind of severe diseases like coronavirus disease 2019, diabetes, and microbial infections as well as individuals undergoing cancer diagnosis. Currently, commercial methods employed for platelet level determination in whole-blood are based on Coulter principle, flow cytometry, and image analysis. However, these technologies are intricate, time consuming, expensive, and require a trained operator as well as lack of portability. Considering these issues, we have developed a self-powered portable device for real-time platelet level monitoring, aimed towards point-of-care applications. The fabricated self-powered-microfluidic triboelectric nanogenerator (SPM-TENG) utilized test sample flow-resistance for sensing of platelet level and triboelectric voltage output as sensing signal. For the fine tuning of fabricated SPM-TENG, we compared the triboelectric voltage output from gold electrode measurement setup and terminal electrode measurement setup. In view of further improvement in stability, voltage output and to minimize the noise in sensing signals, the geometrical optimization of microfluidic channel was performed. In conclusion we have optimized the device as a terminal electrode measurement setup with copper tubes on terminals of microfluidic channel and copper oxide nanowires grown on the interior of copper tubes. These copper oxide nanowires provide a high surface area for solid-liquid contact separation and is responsible for enhanced sensing signal. The fabricated SPM-TENG has shown its great potential in platelet level sensing with plasma-based samples as well as whole-blood samples. The proposed device demonstrated its practicality towards real-time rapid sensing of platelet level and portability.