Ultrasonography is a convenient and widely used technique to look into the longitudinal muscle motion as it is radiation-free and real-time. The motion of localized parts of the muscle, disclosed by ultrasonography, spatially reflects contraction activities of the corresponding muscles. However, little attention was paid to the estimation of longitudinal muscle motion, especially towards estimation of dense deformation field at different depths under the skin. Yet fewer studies on the visualization of such muscle motion or further clinical applications were reported in the literature. A primal-dual algorithm was used to estimate the motion of gastrocnemius muscle (GM) in longitudinal direction in this study. To provide insights into the rules of longitudinal muscle motion, we proposed a novel framework including motion estimation, visualization and quantitative analysis to interpret synchronous activities of collaborating muscles with spatial details. The proposed methods were evaluated on ultrasound image sequences, captured at a rate of 25 frames per second from eight healthy subjects. In order to estimate and visualize the GM motion in longitudinal direction, each subject was asked to perform isometric plantar flexion twice. Preliminary results show that the proposed visualization methods provide both spatial and temporal details and they are helpful to study muscle contractions. One of the proposed quantitative measures was also tested on a patient with unilateral limb dysfunction caused by cerebral infarction. The measure revealed distinct patterns between the normal and the dysfunctional lower limb. The proposed framework and its associated quantitative measures could potentially be used to complement electromyography (EMG) and torque signals in functional assessment of skeletal muscles. ; Interdisciplinary Division of Biomedical Engineering