In order to ensure the parallelism between the optical axis of dove prism and its rotation axis, a method based on the reversing image theory of Dove prism and the optical alignment machining was described. This method takes advantage of the optical property that the direction of the output beam changes with the rotation axis diversion of Dove prism to determine the optical axis of Dove prism, and ensures the parallelism between the rotation axis of cylinder with the optical axis of Dove prism by using metal-turning. Two methods which are used to determine the optical axis of Dove prism was proposed. These two methods can ensure the parallelism between the reflecting surface axis of dove prism and its rotating axis, and the accuracy of axis determining is better than 0.01 mm.