The precise measurement and analysis of human movements is an essential step in biomechanical research used in sports or medicine. Measurement systems used for motion tracking should be non-invasive, safe to use, widely customisable and cost-efficient. In this study, complete design, development and evaluation of a high-speed optical motion tracking and analysis system is described. The system aims to analyse movements for sports and medical applications. The novelty of the proposed system is its design, which is based on visible light light-emitting diode (LED) markers, rather than infrared markers that are commonly used, and a pair of high-speed digital cameras. Calibration procedures and a super-resolution marker model are introduced, ensuring sub-pixel marker centre detection which results in higher three-dimensional reconstruction accuracy. Evaluation of the system included an accuracy test of the proposed system on static and moving objects with known dimensions, followed by analysis of kinematic data obtained in dynamic conditions while measuring human gait. The evaluation results are presented, and conclusions about system performance with possible improvements are discussed.