Estimation of force output generated by human muscle is an essential routine of clinical rehabilitation assessment, and could provide considerable insight into rehabilitation, motor control and robotics. Indirect methods for the estimation of force output could be helpful when a bulky and expensive dynamometer is not on hand. Electromyography has been used in previous studies to quantify it in the literature. However, the force output is a summation of the motor unit action potentials, and thus the contributions and performances of superficial and deep-lying muscles could hardly be separated from each other. In this preliminary study, we applied ultrasonography (US) to explore the feasibility of estimating triceps surae force output during isometric plantar flexion with spatial resolution from superficial to deeper muscles. The local deformations of US images are extracted to represent the morphological changes during force generation. It was found US could be utilized to decently (coefficient of determination at 0.875 ± 0.051 and normalized root mean square error 0.160 ± 0.035) estimate the force output and the measured force by a dynamometer.