Here, we describe the development of chloroplasts and the buildup of the thylakoid membranes in growing Arabidopsis leaves. Organelles were analyzed from three distinct positions, namely, at the tip, the upper leaf margin, and the base from leaves 1, 3, 5, and 7 of 14-day-old plants. Clear developmental gradients are described within a given leaf and between leaves of different age. Chloroplasts at the tip of every leaf are always the most matured within a given leaf, while already at the upper leaf margin a differentiation gradient can be observed from the edge of the leaf toward the midrib. The data presented here can serve as a standard for a subcellular phenotypic analysis in chloroplast biogenesis mutants.