AbstractThis study deals with regional differences in the architectural design of the human masseter muscle. For a number of defined muscle regions the three‐dimensional corrdinates of origin and insertion points, and the lengths of the muscle fibers and the sarcomeres were determined in the closed jaw position. Measurements were made from cadavers and the data were used as input for a model predicting sarcomere length at other mandibular positions. At a closed jaw average muscle fiber length of the muscle regions ranged between 19.0–30.3 mm. The fibers appeared to be considerably longer (35%) anteriorly than posteriorly in the muscle, and deeply situated fibers were on average 5% shorter than superficially situated ones. Average sarcomere length of the regions ranged between 2.27–2.55 μm, indicating that at a closed jaw position sarcomeres are at suboptimum length and have different positions on the length‐tension curve. In the deep layer of the muscle sarcomeres were significantly shorter (6%) than in the superficial layer. Within the superficial layer sarcomere lengths did not differ significantly, but in the deep layer sarcomeres were shorter (8%) posteriorly than anteriorly in the muscle. The model shows that jaw displacement had a different effect on sarcomere length in the muscle regions. When the jaw was rotated about a transverse axis (open/close rotation) sarcomere excursions were relatively small in the posterior muscle regions and large in the anterior regions. The reverse was true when the jaw was rotated contralaterally about a vertical axis. It is concluded that, due to heterogeneity in fiber and sarcomere lengths, the distribution of maximal isometric tension across the muscle at full effort is not