In order to calculate the mixed flow capacity of non-motorized lanes more reasonably based on the characteristics of staggered driving and serpentine movement of non-motorized vehicles, a method of calculating the mixed flow capacity of non-motorized lanes based on time-space consumption was proposed. The mixed traffic capacity of seven non-motorized lanes in Wuhan is calculated by using the observed traffic flow data during the saturation hours of these seven non-motorized lanes. The effects of non-motorized vehicle speed and the proportion of e-bikes on the average time-space consumption of a single non-motorized vehicle under different isolation method and widths were obtained using data fitting and variance analysis, and then the mixed traffic capacity characteristics of non-motorized lanes are analyzed. The results of the case study show that: the average time-space consumption of pure e-bikes is about 2.15 (m²·s)/veh lower than that of a single non-motorized vehicle with pure manual bikes, and the operating efficiency of e-bikes is higher than that of manual bikes, which has an enhancing effect on the capacity. The calculation and analysis of mixed non-motorized lane capacity based on time-space consumption can overcome the drawbacks of ignoring non-motorized vehicle driving characteristics and multi-factor simplification to reduce the accuracy of non-motorized lane mixed flow capacity calculations, and the calculation principle is clear and easy to carry out on-road tests.