An optical sensing module is configured to detect a characteristic of a sample. The optical sensing module includes a light source, a light guide plate, a first cladding layer, a light converging layer, a filter layer, and a plurality of sensors. The light source is configured to provide an exciting beam. Positions of the sensors correspond to positions of the holes. After the exciting beam enters the light guide plate, at least one portion of the exciting beam is transmitted to the sample through a portion of the surface of the light guide plate exposed by the holes, the sample is excited by the exciting beam to emit a signal beam, and the signal beam passes through the light converging layer and the filter layer in an order and travels to the sensors. Another optical sensing module is also provided.