Three-dimensional (3D) H-scan ultrasound (US) is a new high-resolution imaging technology for voxel-level tissue classification. For the purpose of validation, a simulated H-scan US imaging system was developed to comprehensively study the sensitivity to scatterer size in volume space. A programmable research US system (Vantage 256, Verasonics Inc, Kirkland, WA) equipped with a custom volumetric imaging transducer (4DL7, Vermon, Tours, France) was used for US data acquisition and comparison to simulated findings. Preliminary studies were conducted using homogeneous phantoms embedded with acoustic scatterers of varying sizes (15, 30, 40 or 250 μm). Both simulation and experimental results indicate that the H -scan US imaging method is more sensitive than B-mode US in differentiating US scatterers of varying size. Overall, this study proved useful for evaluating H -scan US imaging of tissue scatterer patterns and will inform future technology research and development.