The study compared changes in intramuscular and surface recordings of EMG amplitude with ultrasound measures of muscle architecture of the elbow flexors during a submaximal isometric contraction. Ten subjects performed a fatiguing contraction to task failure at 20% of maximal voluntary contraction (MVC) force. EMG activity was recorded in biceps brachii, brachialis, and brachioradialis muscles using intramuscular and surface electrodes. The rates of increase in the amplitude of the surface EMG for the long and short heads of biceps brachii and brachioradialis were greater than those for the intramuscular recordings measured at different depths. The amplitude of the intramuscular recordings from three muscles increased at a similar rate (P = 0.13), as did the amplitude of the three surface recordings from two muscles (P = 0.83). The increases in brachialis thickness (27.7 ± 5.7 to 30.9 ±3.5 mm, P < 0.05) and pennation angle (10.9 ± 3.5 to 16.5± 4.8 degrees, P = 0.003) were not associated with the increase in intramuscular EMG amplitude (P > 0.58). The increase in brachioradialis thickness (22.8 ± 4.8 to 25.5 ± 3.4 mm, P = 0.0075) was associated with the increase in the amplitude for one of two intramuscular EMG signals (P = 0.007, r = 0.79). Time to failure was more strongly associated with the rate of increase in the amplitude of the surface EMG than that for the intramuscular EMG, which suggests that the surface measurement provides a more appropriate measure of the change in muscle activation during a fatiguing contraction.