Muscle morphological parameters such as fascicle length (FL), pennationangle (PA) and physiologic cross-sectional area (PCSA) can provide an insightinto the reasons of the deteriorated muscle functions caused by pathologies.This study investigates the 3D structure of the lower leg muscles using 3Dfreehand ultrasound (3DfUS). This imaging modality uses a motion capturesystem to track the position of the US probe during acquisition and thusreconstruct the structure of the tissues in 3D. In this study, two subjects werescanned on the medial gastrocnemius (MG) and tibialis anterior (TA) musclesin the lower leg using 3DfUS system. The FL and PA of the muscles werecalculated and compared with the values previously measured using diffusiontensor imaging (DTI). The results using 3DfUS were averagely 19.2% largerin FL and 2.9%larger in PA. In conclusion, 3DfUS can successfully determinemuscle morphological parameters within a physiologically acceptable range.But the differences in FL observed between the two imaging modalities werequite big, which probably was due to the differences in sample size and area.The values can also differ greatly within the 3DfUS measurements as a resultof different manipulations during data processing, and the 3DfUS protocolneeds to be further improved in future studies.