This study aimed (i) to investigate the role of regional characteristics of the knee extensors muscles (vastus lateralis: VL, vastus intermedius: VI and rectus femoris: RF) in determining maximum-voluntary force (MVF); and (ii) to understand which regional parameter of muscle structure would best predict MVF. Muscle architecture (e.g., pennation angle and fascicle length), muscle volume (Vol), anatomical (ACSA) and physiological cross-sectional-area (PCSA) were measured in the proximal (0–33% of the muscle length), middle (33–66% of the muscle length) and distal (66–100% of the muscle length) portions of each muscle in fifteen healthy males using ultrasound and Magnetic Resonance Imaging (MRI). Knee extensors force was calculated in isometric condition at a single knee joint angle of 90 degrees. Regional ACSA, Vol and PCSA were correlated with MVF production. Regional muscle geometry showed no significant correlations with MVF. Among regions, the middle portion of each muscle was largely correlated with MVF compared to all the other regions (distal and proximal). To understand which regional structural parameter best predicted MVF, a stepwise multiple linear regression was performed. This model showed a significant explanatory power (P < 0.001, R2 = 0.76, adjusted R2 = 0.71), including muscle Vol collected in the mid portions of VL and RF. Even if no significant differences were reported between Vol, PCSA and ACSA in determining MVF, our results showed that the RF and VL volume collected in the middle portion of the muscle length are strong determinants of MVF produced by the knee extensors at 90 degrees joint angle.