The normal function of the lateral pterygoid muscle is not well understood although this muscle is thought to play an important role in the control of jaw and jaw-joint function and is implicated in temporomandibular disorders (TMD). The lack of a validated method for standardization of jaw movement in studies of lateral pterygoid function has contributed to the lack of understanding of the normal function of this muscle. An improved understanding of normal function will allow valid comparisons to be made with TMD patients in order to identify whether purported differences in activity actually exist. This paper describes a methodology for standardizing command jaw movements in the horizontal plane, together with reliable recordings of single-motor-unit (SMU) activity. In six human participants, jaw movements were standardized by having participants track a linear bank of light-emitting diodes (LEDs) aligned on a monitor displaying the mid-incisor point (MIPT). In all participants, the MIPT target (i.e. an illuminated LED) could be tracked, according to a pre-determined criterion, during single- and multiple-step displacements at different rates (1.3–6.5 mm/s at MIPT) and magnitudes (0.65–12 mm) of movement. SMU activity from the superior (SHLP) or inferior (IHLP) head of the lateral pterygoid muscle could be reliably discriminated during repeated trials of these defined tasks. This methodology establishes a reliable technique for characterizing the firing properties of SMUs within the lateral pterygoid, and has implications for analogous studies in other jaw muscles.