Instrumented treadmills are increasingly used in gait research, although the imposed walking speed is suggested to affect gait performance. A feedback-controlled treadmill that allows subjects to walk at their preferred speed, i.e. functioning in a self-paced (SP) mode, might be an attractive alternative, but could disturb gait through accelerations of the belt. We compared SP with fixed speed (FS) treadmill walking, and also considered various feedback modes. Nineteen healthy subjects walked on a dual-belt instrumented treadmill. Spatio-temporal, kinematic and kinetic gait parameters were derived from both the average stride patterns and stride-to-stride variability. For 15 out of 70 parameters significant differences were found between SP and FS. These differences were smaller than 1. cm, 1°, 0.2. N. m and 0.2. W/kg for respectively stride length and width, joint kinematics, moments and powers. Since this is well within the normal stride variability, these differences were not considered to be clinically relevant, indicating that SP walking is not notably affected by belt accelerations. The long-term components of walking speed variability increased during SP walking (43%, p< 0.01), suggesting that SP allows for more natural stride variability. Differences between SP feedback modes were predominantly found in the timescales of walking speed variability, while the gait pattern was similar between modes. Overall, the lack of clinically significant differences in gait pattern suggests that SP walking is a suitable alternative to fixed speed treadmill walking in gait analysis. © 2013 Elsevier B.V.