Lipofuscin is one of the indicators of oxidative stress. To elucidate the role of oxidative stress in the development of wooden breast, this study investigates lipofuscin accumulation in various parts of wooden breast muscles. Sampling was performed using 46-day-old broiler chickens housed at a commercial Japanese poultry slaughterhouse. Fourteen wooden breast fillets and 13 normal breast fillets were collected in the deboning line. The samples used to measure shear force, 2-thiobarbituric acid reactive substance (TBARS), and for histological analysis were taken from the six portions of breast muscle fillets. In muscles affected by wooden breast, vacuolated muscle fibers were observed, and connective tissues appearing like perimysium were expanded with fibrosis. TBARS value and accumulation of lipofuscin were significantly higher in the wooden breast than in the normal breasts. A lot of lipofuscin granules were localized in the cytoplasm of collapsed muscle fibers of the wooden breast. The cranial portion of the wooden breast showed the highest shear force. The cranial position had a large amount of connective tissue and lipofuscin granules. The results of the present study strongly suggest that high oxidative stress, especially with a significant accumulation of lipofuscin, is associated with the development of wooden breasts.