Chicks were injected intra-cerebrally with [(3)H]leucine, and (3)H-labeled microtubules were assembled in vitro in the brain supernatants. Pieces of these (3)H-labeled microtubules were then incubated with unlabeled brain tubulin subunits under conditions where the subunits assembled onto the labeled pieces. Electron microscopic autoradiography of the negatively-stained microtubules showed all of the radioactivity at one end of the tubules as they increased in length. This clearly demonstrated that the microtubules of brain in this in vitro system were being assembled unidirectionally.