Particle Image Velocimetry (PIV) is an important technique in studying blood flow in heart valves. Previous PIV studies of flow around prosthetic heart valves had different research concentrations, and thus never provided the physical flow field pictures in a complete heart cycle, which compromised their pertinence for a better understanding of the valvular mechanism. In this study, a digital PIV (DPIV) investigation was carried out with improved accuracy, to analyse the pulsatile flow field around the bi-leaflet mechanical heart valve (MHV) in a complete heart cycle. For this purpose a pulsatile flow test rig was constructed to provide the necessary in vitro test environment, and the flow field around a St. Jude size 29 bi-leaflet MHV and a similar MHV model were studied under a simulated physiological pressure waveform with flow rate of 5.21/min and pulse rate at 72 beats/min. A phase-locking method was applied to gate the dynamic process of valve leaflet motions. A special image-processing program was applied to eliminate optical distortion caused by the difference in refractive indexes between the blood analogue fluid and the test section. Results clearly showed that, due to the presence of the two leaflets, the valvular flow conduit was partitioned into three flow channels. In the opening process, flow in the two side channels was first to develop under the presence of the forward pressure gradient. The flow in the central channel was developed much later at about the mid-stage of the opening process. Forward flows in all three channels were observed at the late stage of the opening process. At the early closing process, a backward flow developed first in the central channel. Under the influence of the reverse pressure gradient, the flow in the central channel first appeared to be disturbed, which was then transformed into backward flow. The backward flow in the central channel was found to be the main driving factor for the leaflet rotation in the valve closing process. After the valve was fully closed, local flow activities in the proximity of the valve region persisted for a certain time before slowly dying out. In both the valve opening and closing processes, maximum velocity always appeared near the leaflet trailing edges. The flow field features revealed in the present paper improved our understanding of valve motion mechanism under physiological conditions, and this knowledge is very helpful in designing the new generation of MHVs.