Ankle plantar flexor muscles support and propel the body in the stance phase of locomotion. Besides the triceps surae, flexor hallucis longus muscle (FHL) may also contribute to this role, but very few in vivo studies have examined FHL function during walking. Here, we investigated FHL fascicle behavior at different walking speeds. Ten healthy males walked overground at three different speeds while FHL fascicle length changes were recorded with ultrasound and muscle activity was recorded with surface electromyography (EMG). Fascicle length at heel strike at toe off and at peak EMG activity did not change with speed. Range of FHL fascicle length change (3.5-4.5 and 1.9-2.9 mm on average in stance and push-off phase, respectively), as well as minimum (53.5-54.9 and 53.8-55.7 mm) and maximum (58-58.4 and 56.8-57.7 mm) fascicle length did not change with speed in the stance or push-off phase. Mean fascicle velocity did not change in the stance phase, but increased significantly in the push-off phase between slow and fast walking speeds (P=.021). EMG activity increased significantly in both phases from slow to preferred and preferred to fast speed (P<.02 in all cases). FHL muscle fascicles worked near-isometrically during the whole stance phase (at least during slow walking) and operated at approximately the same length at different walking speeds. FHL and medial gastrocnemius (MG) have similar fiber length to muscle belly length ratios and, according to our results, also exhibit similar fascicle behavior at different walking speeds.