A method of laser cubic prism collimation and attitude measurement was proposed in this paper, and the accuracy was analyzed. A single cubic prism and double cubic prisms were collimated and measured respectively by using high precision electronic theodolites and laser tracker system. The angle of adjacent surface and coordinate transformation parameters were calculated, and the measuring accuracy of new method was compared. The measurement efficiency and environmental requirements was also compared. The experiments results show that the new method achieved an accuracy of 10″, which is significantly inproved compared to other traditional methods with an accuracy of 0.5′, indicating that this new method had the equivalent accuracy to other mainstream theodolite method. At the same time the efficiency of new method was improved by more than double, and the requirements for the measurement environment were also more relaxed. It could replace the method of theodolite in the actual production.