A 2-frame PTV (particle tracking velocimetry) system using the concept of match probability between two consequent image frames has been developed to obtain instantaneous velocity fields. The overall 2-frame PTV system including image pre-processing, tracking algorithm and post-processing routine was implemented to apply to real flows. The developed 2-frame PTV system has several advantages such as high recovery ratio of velocity vectors, low error ratio and small computational time compared with the conventional 4-frame PTV and the FFT-based cross-correlation PIV technique. The 2-frame PTV system was applied to a turbulent channel flow over a rectangular block to check its reliability and usefulness. Total 96 sequential image frames have been captured and processed to get both mean and fluctuating velocity vector fields over the recirculating region. The mean velocity and turbulent intensity profiles were well agreed with hte LDV measurements in the separated region behind the block. Time-averaged reattachment length is about 6.3 times of the block height.