The aim of the study was to investigate amplitude and frequency content of single motor unit (MU) electromyographic (EMG) and mechanomyographic (MMG) responses. Multi-channel surface EMG and MMG signals were detected from the dominant biceps brachii muscle of 10 volunteers during isometric voluntary contractions at 20%, 50%, and 80% of the maximal voluntary contraction (MVC) force. Each contraction was performed three times in the experimental session which was repeated in three non-consecutive days. Single MU action potentials were identified from the surface EMG signals and their times of occurrence used to trigger the averaging of the MMG signal. At each contraction level, the MUs with action potentials of highest amplitude were identified. Single MU EMG and MMG amplitude and mean frequency were estimated with normalized standard error of the mean within subjects (due to repetition of the measure in different trials and experimental sessions) smaller than 15% and 7%, respectively, in all conditions. The amplitude of the action potentials of the detected MUs increased with increasing force (mean +/- SD, 244 +/- 116 microV at 20% MVC, and 1426 +/- 638 microV at 80% MVC; P < 0.001) while MU MMG amplitude increased from 20% to 50% MVC (40.5 +/- 20.9 and 150 +/- 88.4 mm/s(2), respectively; P<0.001) and did not change significantly between 50% and 80% MVC (129 +/ -82.7 mm/s(2) at 80% MVC). MU EMG mean frequency decreased with contraction level (20% MVC: 97.2 +/- 13.9 Hz; 80% MVC: 86.2 +/- 11.4 Hz; P < 0.001) while MU MMG mean frequency increased (20% MVC: 33.2 +/- 6.8 Hz; 80% MVC: 40.1 +/- 6.1 Hz; P < 0.001). EMG peak-to-peak amplitude and mean frequency of individual MUs were not correlated with the corresponding variables of MMG at any contraction level.