To address the problems inherent in manual methods, a novel, semi-automated tissue segmentation image analysis technique has been developed. The purpose of this study was to demonstrate the feasibility and describe preliminary observations of applying this technique to quantify and monitor longitudinal changes in abdominal adipose tissue and thigh muscle volume in obese adults during weight loss. Abdominal adipose tissue and thigh muscle volume decreased during weight loss. As a proportion of body weight, adipose tissue volumes decreased during weight loss. By comparison, as a proportion of body weight, thigh muscle volume increased.