Eccentric exercise affects muscles differentially according to intensity, duration, and previous exposure to the specific exercise activity. We used T2-weighted magnetic resonance imaging sequences to localize and quantify muscle damage following step exercise and to determine correlations between transverse relaxation time (T2) and other markers of muscle damage. Eight women performed two-step exercise bouts (30 min) separated by 8 weeks. Blood samples, MR scans, measurements of muscle strength, and muscle soreness were obtained immediately before, after, and up to 9 days after each bout. Resting muscle T2 (40.3+/-0.6 ms) increased exclusively in m. Adductor magnus (AM) in the thigh performing eccentric contractions and peaked 3 days after bout 1 (73.5+/-9.7 ms, P<0.05). Plasma creatine kinase (CK) activity peaked on day 3 after bout 1 and correlated with T2 in AM (r=0.96, P<0.001). After bout 2 CK and T2 were almost unaffected. This indicates that T2-weighted MRI can be applied to identify muscles from which enzymes are being released into the circulation.