The invention discloses an on-orbit self-calibration device of a satellite-borne laser communication machine and a calibration method of the on-orbit self-calibration device. Comprising a laser, an optical fiber isolator, an optical switch, a first optical fiber coupler, a communication receiving end, a receiving optical fiber, a receiving lens group, a beam splitter, a reflector, a tracking and pointing detector, a tracking and pointing lens group, a color separation film, a first fast reflecting mirror, a secondary mirror, a primary mirror, a two-dimensional oscillating mirror, a pyramid, a second fast reflecting mirror, a transmitting lens group, a transmitting optical fiber and a high-power amplifier. And a second optical fiber coupler and a communication transmitting terminal. According to the invention, the optical switches are sequentially turned on to the receiving optical path or the transmitting optical path, and then are reflected to the receiving optical path through the pyramid to detect the optical power, and the tracking and pointing optical path is used for detecting and compensating the mass center position of a light spot, so that the coaxial self-calibration of the receiving optical path, the transmitting optical path and the tracking and pointing optical path is realized. The tracking and pointing precision and the link transmission efficiency are improved, and the debugging difficulty and accuracy of the laser communication machine during satellite in-orbit operation are reduced.