This study examined the effect of pedal cadence on the heterogeneity of muscle deoxygenation during exercise of moderate intensity. Twelve healthy subjects performed 6 min of cycling at 40 and 100 r · min(-1) at 80% of the workload corresponding to the gas exchange threshold. Gas exchanges were measured breath by breath during each exercise. Muscle deoxygenation (HHb, i.e., O2 extraction) was monitored continuously by near-infrared spectroscopy at eight sites on the vastus lateralis. The heterogeneity of HHb was assessed using the relative dispersion of the signal measured at the eight sites (i.e., 100 × standard deviation/mean). HHb was not altered by the pedal cadence, whereas pulmonary V̇O2 was higher at 100 r · min(-1) than at 40 r · min(-1) (p < 0.001). The relative dispersion of HHb was significantly higher at 100 r · min(-1) than at 40 r · min(-1) (p < 0.001). These results indicate that pedal cadence has no effect on O2 extraction but that an elevated cadence would increase muscle V̇O2, suggesting an increase in muscle blood flow. Elevated cadence also induced greater heterogeneity of the muscle's V̇O2/Q̇O2 delivery ratio, suggesting a change in the adequacy between O2 demand and O2 delivery in some regions of active muscle.