This paper presents a novel biosensor for bitter substance detection on the basis of light addressable potentiometric sensor (LAPS). Taste receptor cells (TRCs) were used as sensitive elements, which can respond to different bitter stimuli with extreme high sensitivity and speci- ficity. TRCs were isolated from the taste buds of rats and cultured on the surface of LAPS chip. Due to the unique advantages such as single-cell recording, light addressable capability, and noninvasiveness, LAPS chip was used as secondary transducer to monitor the responses of TRCs by recording extracelluar potential changes. The results indicate LAPS chip can effectively record the responses of TRCs to different bitter substances used in this study in a real-time manner for a long-term. In addition, by performing principal component analysis on the LAPS recording data, different bitter substances tested can be successfully discriminated. It is suggested this TRCs-LAPS hybrid biosensor could be a valuable tool for bitter substance detection. With further improvement and novel design, it has great potentials to be applied in both basic research and practical applications related to bitter taste detection.