The main objective of this thesis is to evaluate accuracy and precision of the machine vision system used to calibrate a prepared environment foroptical navigation. Rotationally independent optimized colour reference labels (symbols) creates an environment. Any number of symbols can be used. A symbol carries 8–bit (0 to 255) information, which can be designed for different values by using Matlab algorithms. An optical navigation system enters into the environment and captures thesymbols. The symbols are then decoded to determine the geographical positions of the symbols from reference position of the system by using Matlab algorithms. Then, the system is moved to a known position and the same set of symbols are captured, decoded and located. The process is repeated for several positions of the system to find precision and accuracy. Finally, the results are analysed.