Tibiae from 11-day-old chick embryos, injected with 3H-tetracycline, were autoradiographically analyzed at different stages to localize and study the dynamics of the initial phases of bone mineralization, 3H-tetracycline was localized within newly formed trabeculae, but only at the surfaces of older trabeculae, indicating that the incorporation of tetracycline into bone occurs at active sites of calcification. It takes between 24 and 36 h for injected tetracycline to become stabilized and incorporated into the mineralized matrix. Absence of 3H-tetracycline grains over the osteoblasts suggests a paracellular pathway for incorporation of tetracycline into the mineralizing bone matrix.