High-resolution facial surface electromyography (HR-sEMG) is suited to discriminate between different facial movements. Whether HR-sEMG also allows a discrimination among the six basic emotions of facial expression is unclear. 36 healthy participants (53% female, 18–67 years) were included for four sessions. Electromyograms were recorded from both sides of the face using a muscle-position oriented electrode application (Fridlund scheme) and by a landmark-oriented, muscle unrelated symmetrical electrode arrangement (Kuramoto scheme) simultaneously on the face. In each session, participants expressed the six basic emotions in response to standardized facial images expressing the corresponding emotions. This was repeated once on the same day. Both sessions were repeated two weeks later to assess repetition effects. HR-sEMG characteristics showed systematic regional distribution patterns of emotional muscle activation for both schemes with very low interindividual variability. Statistical discrimination between the different HR-sEMG patterns was good for both schemes for most but not all basic emotions (ranging from p > 0.05 to mostly p < 0.001) when using HR-sEMG of the entire face. When using information only from the lower face, the Kuramoto scheme allowed a more reliable discrimination of all six emotions (all p < 0.001). A landmark-oriented HR-sEMG recording allows specific discrimination of facial muscle activity patterns during basic emotional expressions.