The invention discloses a high-resolution biosensor. Specifically, a basic unit consisting of a first insulating layer, a nanometer functional layer and a second insulating layer is provided with a nano-pore in the center, thus forming a nanometer functional layer unit. A first electrophoresis electrode or micropump, a first store room, a second store room, a second electrophoresis electrode or micropump, and a micro-nanometer separation channel constitute a micro-nanometer fluid device unit. The nanometer functional layer unit, a source electrode, a drain electrode, a dielectric layer and a gate electrode compose a field effect transistor unit. When a biomolecule passes through the nano-pore in the micro-nanometer fluid device and interacts with the nanometer functional layer, the field effect transistor unit measures the field effect characteristic change resulted from the interaction, thus reaching the purpose of biomolecule detection. The invention provides a solution to the technical difficulty of integrating the nano-pore in the nanometer functional layer, and can control the form change when a biomolecule passes through the nano-pore, thus meeting the resolution requirements for detecting the biomolecule characteristic and structure. In addition, the manufacturing method of the sensor is simple.