A wireless multi-layered sensor that allows electromyography (EMG), mechanomyography (MMG) and near-infrared spectroscopy (NIRS) measurements to be carried out simultaneously is presented. The multi-layered sensor comprises a thin silver electrode, transparent piezo-film and photosensor. EMG and MMG measurements are performed using the electrode and piezo-film, respectively. NIRS measurements are performed using the photosensor. Muscular activity is then analyzed in detail using the three types of data obtained. In experiments, the EMG, MMG and NIRS signals were measured for isometric ramp contraction at the forearm and cycling exercise of the lateral vastus muscle with stepped increments of the load using the layered sensor. The results showed that it was possible to perform simultaneous EMG, MMG and NIRS measurements at a local position using the proposed sensor. It is suggested that the proposed sensor has the potential to evaluate muscular activity during exercise, although the detection of the anaerobic threshold has not been clearly addressed.