ObjectiveThe present study evaluated the in vivo musculoaponeurotic architecture of the superficial head (SH) of the masseter muscle (MM) of asymptomatic participants in excursive mandibular movements compared to the relaxed state as examined with ultrasound imaging. It was hypothesized that the mean fiber bundle length (FBL) and mean height of the aponeurosis (HA) of the laminae of the SH would differ significantly between the relaxed state and protrusion, ipsilateral excursion, and contralateral excursion. Study DesignThe MM was studied volumetrically in 12 female and 12 male asymptomatic participants bilaterally by using ultrasound imaging. Mean FBL and HA in protrusion and ipsilateral and contralateral excursion were compared to these values in the relaxed state using paired t-tests (P < .05). The intraclass correlation coefficient was used to assess intraexaminer reliability. ResultsThe SH exhibited multiple laminae. Fiber bundles were found to attach to bone and the superior and inferior aponeuroses. Mean FBL was significantly shorter and mean HA significantly longer in protrusion and the excursions than in the relaxed state although the pattern of altered laminae and aponeuroses differed among the mandibular movements. Intraexaminer reliability was excellent. ConclusionSpecific changes in mean FBL and mean HA suggest differential contraction of the SH of the MM based on laminar morphology. These findings provide a baseline to investigate musculoaponeurotic changes in patients with myogenic masseter muscle pain.