The aim of the present study was to determine the validity of ultrasound (US) imaging versus magnetic resonance imaging (MRI) for measuring anterior thigh muscle, subcutaneous adipose tissue (SAT), and fascia thickness. Twenty healthy, moderately active participants (aged 49.1 ± 9.74 [36–64] years), underwent imaging of the anterior thigh, using ultrasound and MRI modalities on the same day. Images were analyzed offline to assess the level of agreement between US and MRI measurements. Pearson’s correlation coefficient showed an excellent relationship between US imaging and MRI for measuring muscle (r = 0.99, p < 0.01), SAT (r = 0.99, p < 0.01), and non-contractile tissue (SAT combined with perimuscular fascia) thickness (r = 0.99, p < 0.01). Perimuscular fascia thickness measurement showed a poor correlation between modalities (r = 0.39, p < 0.01). Intra-class correlation coefficients (ICC3,1) also showed excellent correlation of the measurements with ICC = 0.99 for muscle thickness, SAT, and non-contractile tissue, but not for perimuscular fascia, which showed poor agreement ICC = 0.36. Bland and Altman plots demonstrated excellent agreement between US imaging and MRI measurements. Criterion validity was demonstrated for US imaging against MRI, for measuring thickness of muscle and SAT, but not perimuscular fascia alone on the anterior thigh. The US imaging technique is therefore applicable for research and clinical purposes for muscle and SAT.