Calcein entrapped ethosomes, which are vesicular system composed mainly of phospholipid, ethanol, and water, were prepared with varying the composition of constituent components and preparation condition. Entrapment efficiency of the ethosome is decreased as the added amount of ethanol increased. The ethosome should be prepared above 60 ℃ for high entrapment efficiency. Incorporation of cholesterol into lecithin decreased the entrapment efficiency and increased the droplet size of the ethosome.