This paper presents a high resolution measurement set-up developed for calibrating precise line scales with measurement uncertainty of less than 0.1 µm over a total length of 500 mm. The system integrates a numerically controlled multi-axis stage, a laser interferometer, and a vision system for detecting line position. The measurement and the analysis processes are completely automated in order to minimize manual labour during the calibration process, but also increase the calibration accuracy. Increasing calibration accuracy leads up to better quality of industrial measurements which is required by modern precision industry. When designing this set-up, special attention was paid to the alignment of the measurement object in the measurement direction, considering the focus of the camera. The aim of this alignment was to reduce Abbe errors in 2 axes to negligible level. In addition, all uncertainty contributions have been determined and evaluated by performing extended experiments in specific measurement conditions. These contributions are presented in the uncertainty budget. The metrological capabilities of the presented measurement set-up were verified by some practical test measurements. Selected results of these measurements are presented in the article. This set-up will primarily improve a standard base for calibration of optical measuring devices. The use of the optical standards in the industry is constantly growing. Indirect users of the results of this research will be all manufacturers of precise products such as automotive and other industries.