The invention discloses a three-dimensional laser alignment positioner for a particle image velocimetry and belongs to the technical field of fluid dynamic experiments. The positioner comprises a three-dimensional coordinate adjusting frame and a laser sheet optical correction system. The three-dimensional coordinate adjusting frame comprises a Z-direction coordinate frame, a Y-direction coordinate frame, an X-direction coordinate frame, a front positioning cross ruler, a rear positioning cross ruler, a rotating substrate and a base; and the laser sheet optical correction system comprises two upper and lower laser sheet optical correction boards with the same structure, a focusing ruler, a photoelectric detection board and a photoelectric indicator. By using the three-dimensional coordinate adjusting frame, the positioner can accurately determine the position of a testing plane, realizes the overlap of three surfaces of a flow field testing plane, a laser sheet light surface and a camera shooting surface through the laser sheet optical correction system, automatically detects the overlap ratio through a photoelectric detection device, and avoids human errors brought by the conventional visual inspection and other experimental calibration methods. Therefore, the calibration process of the particle image velocimetry is simpler and more standard and accurate.