The purpose of this study was to investigate if force enhancement (FE) in vivo is influenced by stretch-induced changes of muscle architecture. Therefore, 18 subjects performed maximum voluntary isometric (100° knee flexion angle) and isometric-eccentric-isometric stretch contractions (80°–100° ω = 60°s−1) whereby pennation angle and fascicle length of vastus lateralis was determined using ultrasonography. We found significant (2-way repeated ANOVA; α = 0.05) enhanced torque of 5–10% after stretch as well as significant passive FE but no significant differences in muscle architecture between isometric and stretch contractions at final knee angle. Furthermore, EMG recordings during a follow-up study (n = 10) did not show significant differences in activation and mean frequency of contraction conditions. These results indicate that FE in vivo is not influenced by muscle architectural changes due to stretch.