During running at a constant speed, the optimal stride frequency (SF) can be derived from the u-shaped relationship between SF and heart rate (HR). Changing SF towards the optimum of this relationship is beneficial for energy expenditure and may positively change biomechanics of running. In the current study, the effects of speed on the optimal SF and the nature of the u-shaped relation were empirically tested using Generalized Estimating Equations. To this end, HR was recorded from twelve healthy (4 males, 8 females) inexperienced runners, who completed runs at three speeds. The three speeds were 90%, 100% and 110% of self-selected speed. A self-selected SF (SFself) was determined for each of the speeds prior to the speed series. The speed series started with a free-chosen SF condition, followed by five imposed SF conditions (SFself, 70, 80, 90, 100 strides·min-1) assigned in random order. The conditions lasted 3 minutes with 2.5 minutes of walking in between. SFself increased significantly (p<0.05) with speed with averages of 77, 79, 80 strides·min-1 at 2.4, 2.6, 2.9 m·s-1, respectively). As expected, the relation between SF and HR could be described by a parabolic curve for all speeds. Speed did not significantly affect the curvature, nor did it affect optimal SF. We conclude that over the speed range tested, inexperienced runners may not need to adapt their SF to running speed. However, since SFself were lower than the SFopt of 83 strides·min-1, the runners could reduce HR by increasing their SFself.