Purpose. The purpose of the study was to compare the spatio-temporal and joint kinematic gait parameters of stroke patients walking on a treadmill and overground, to examine the assumption that patients walking on a treadmill will approximate the requirements of walking overground. Methods. Ten independently ambulant chronic stroke patients were included in the study. Vicon was used to collect spatio-temporal and joint kinematic data during overground walking at comfortable speed and at matched speed on the treadmill. Results. Walking on the treadmill demonstrated statistically significantly lower cadence, and longer step times of the non-hemiplegic and hemiplegic limbs. Absolute stance times of both limbs, absolute double support time, relative stance time and relative double support time were significantly longer during treadmill walking. Compared to overground walking, the inter-limb symmetries of step time, stance time, and stance/swing time ratio were significantly greater on the treadmill. During treadmill walking, joint kinematic data showed statistically significant changes with greater flexion of the nonhemiplegic knee and hip at initial contact, and less hip extension of the hemiplegic limb. Maximal ankle plantarflexion and knee extension of the hemiplegic limb occurred later in the gait cycle on the treadmill. Conclusion. These differences suggest it may be useful to use treadmill in conjunction with overground walking to focus on improving specific walking deficits in patients with stroke.