A novel non-contact, five-axis measuring machine with high measurement accuracy of workpiece dimensions is introduced in this paper. The kinematic model, as well as the kinematic error model, is developed. A self-calibration method using a steel ball is proposed to improve the measurement accuracy in the workspace. The calibration process is low-budget and easy-to-operate due to the fact there is no need to rely on other instruments or devices except for the laser probe carried by the measuring machine itself. The objective function is defined in terms of center-to-center distance deviation, namely the theoretical ball center and tested ball center, and to improve the speed of the convergence rate and to increase the optimization accuracy of the genetic algorithm. The simulation and practical experiments both illustrate the feasibility and validity of the proposed kinematic model and self-calibration method. Finally, the actual measurement results of a φ 12 sphere illustrate that the measurement accuracy of this machine has improved greatly by using calibrated parameters against nominal parameters.