The lipid and protein components of pulmonary surfactant are synthesized by alveolar type II cells and stored in their secretory granules, the lamellar bodies. Tubular myelin is a highly ordered surfactant structure that is lung specific and produced in the alveolar airspace. Recent work identified Ca2+, surfactant apoproteins A and B, and saturated phospholipids as necessary for tubular myelin structure and function. We have used serial ultrathin sections from fetal rat lungs and reconstructed the three-dimensional structure of tubular myelin. The spheres of tubular myelin we viewed were 2-3 microns in diameter and were surrounded by up to 20 lamellar bodies, each apparently simultaneously contributing material to the tubules. We measured a long-range symmetry of the tubules, which were folded about a central axis and showed a repeated crossing of individual tubes from one side to the other of the large structure. The tubes appeared closed on their outer edges and there was a delay in the penetration of tubules by cationic ferritin added to tissue slices containing the tubular myelin, although within a few minutes tracer appeared in the lumina and on the walls of the tubules. The three-dimensional images were compatible with existing views of tubular myelin.