The multifunction of molecule-based devices is always achieved by improving their charge transport characteristics. These characteristics depend strongly on the energy levels of molecular semiconductors, which fundamentally govern the working principle and device performance. Therefore, an accurate measurement of these energy levels is crucial for evaluating the availability of the prepared materials and thus optimizing the device performance. Here, an easy-to-operate three-terminal hot electron transistor has been developed, which comprises a molecular optoelectronic device that records the charge transport. It achieves exceptional properties including the lowest unoccupied molecular orbit level, highest occupied molecular orbit level, higher energy states, and higher electronic bandgap. When compared with existing techniques such as cyclic voltammetry, inverse photoemission spectroscopy, and ultraviolet photoemission spectroscopy, the hot electron transistor provides in-situ characterization and categorizes the measured energy information as intrinsic properties of the molecular semiconductor. Furthermore, we provide an in-depth understanding of the fundamental device-physics, which provides promising guidance for performance optimization.