The purpose of the present study is to determine whether muscle functional MRI (mfMRI) can be used to obtain three-dimensional (3-D) images useful for evaluating muscle activity, and if so, to measure the distribution of muscle activity within a medial gastrocnemius (MG) muscle. Seven men performed 5 sets of 10 repetitions of a calf-raise exercise with additional 15% of body-weight load. Magnetic resonance images were obtained before and immediately after the exercise. To threshold images, only those pixels showing transverse relaxation time (T2) greater than the mean+1 S.D. of the entire regions of interest (ROIs) in the preexercise image and T2 lower than the mean+1 S.D. of the entire ROIs in the postexercise image were identified. The survived pixels showing T2 are defined as active muscle. Those thresholded images were 3-D reconstructed, and this was used to determine area of active muscle along transverse, longitudinal and vertical axes. At the exercise level used in the present study, the percentage volume of activated muscle in the MG was 62.8+/-4.5%. There was a significant correlation between percentage volume of activated muscle and integrated electromyography (r=.78, P<.05). Percentage areas of activated muscle were significantly larger in the medial than in the lateral region, in the anterior than in the posterior region and in the distal than in the proximal region (P<.05). These results suggest that mfMRI can be used to evaluate the muscle activity and to determine intramuscular variations of activity within skeletal muscle.