Capillary transit time and its heterogeneity have a marked impact on oxygen extraction in different tissues. Animal studies have shown that exercise shortens capillary transit time but the effects on capillary transit time heterogeneity have been controversial. We investigated whether exercise changes muscle blood transit time heterogeneity in humans in vivo and whether this heterogeneity correlates to muscle oxygen extraction. Muscle blood flow, blood volume, and oxygen uptake were measured during rest and low-intensity exercise in 12 healthy men using positron emission tomography (PET). Blood transit time was calculated from parametric PET images voxel by voxel by dividing blood volume with blood flow. Oxygen extraction was calculated by nonlinear fitting from dynamic 15O-O2 data. Relative dispersion (=SD/mean) was calculated as an index of heterogeneity of blood volume and blood transit time. As expected, exercise significantly shortened blood transit time and increased oxygen extraction. Furthermore, exercise decreased transit time heterogeneity (from 47 +/- 9% to 39 +/- 10%, P=0.07). Transit time heterogeneity correlated inversely to oxygen extraction in the exercising (r=-0.76, P=0.004) but not in the resting muscle (r=0.04, P=0.89). These results show that even low-intensity exercise shortens blood transit time markedly and decreases its heterogeneity in human skeletal muscle in vivo. Findings in correlation analyses suggest that less heterogeneous blood transit time associates to better muscle oxygen extraction during exercise. This may have effects on muscle oxygenation during exercise.