The working principles of Laser Demarcation Instruments (LDIs) and their standard errors were analyzed. A new set of Multi-dimensional Calibration System (MDCS) for the LDIs was developed to solve these shortcomings of traditional calibration systems on larger floor spaces, poor accuracy and high influential artificial factors. A solution based on collimator angle measurement and machine vision measurement was designed to calibrate the LDI by measuring the straightness and verticality of laser lines. The data capturing platform of the system consisted of 8 collimators was used to achieve the multi-dimensional calibration. The software was designed by mixed programming of VC++ and Matlab, and they were connected by calling a Dynamic Lined Library(DLL). Experiments show that the new system is easy to operate, and is more efficient and more credible. The space that the whole system takes up is within 8 m3, which saves the area of work place. Moreover, the accuracy of horizontal and vertical lines can reach ±0.2 mm/5 m, and the orthogonal line is ±23″, which meets the requirement of calibration accuracy.