The objective of this study was to systematically characterize motor units (MUs) of the musculature of the lower face. MU endplate positions and principal muscle fiber orientations relative to facial landmarks were identified. This was done by the analysis of motor unit action potentials (MUAPs) in the surface electromyogram. Thirteen specially trained, healthy subjects performed selective contractions of the depressor anguli oris, depressor labii inferioris, mentalis, and orbicularis oris inferior muscles. Signals were recorded using recently developed, 0.3-mm thin and flexible high-density surface electromyography (sEMG) grids (120 channels). For each subject and each muscle and for different low contraction levels, representative MUAPs ("MU fingerprints") were extracted from the raw sEMG data according to their spatiotemporal amplitude characteristics. We then topographically characterized the lower facial MUs' endplate zones and main muscle fiber orientations on the individual faces of the subjects. These topographical MU parameters were spatially warped to correct for the different sizes and shapes of the faces of individual subjects. This electrophysiological study revealed a distribution of the lower facial MU endplates in more or less restricted, distinct clusters on the muscle often with eccentric locations. The results add substantially to the basic neurophysiologic and anatomical knowledge of the complex facial muscle system. They can also be used to establish objective guidelines for placement of conventional (surface or needle) EMG electrodes as well as for clinical investigations on neuromuscular diseases affecting the facial musculature. The localized endplate positions may also indicate optimal locations for botulinum toxin injection in the face.