A method of manufacturing a hollow micro-needle structure includes the steps of: disposing a first mask layer and a second mask layer respectively aside a first substrate and aside a rear surface of the first substrate, wherein the first substrate is transparent to predetermined light; forming a photoresist layer on the front surface of the first substrate and the first mask layer; providing the predetermined light to illuminate the first substrate in a direction from the rear surface to the front surface so as to expose the photoresist layer to form an exposed portion and an unexposed portion; and removing the unexposed portion to form the micro-needle structure, which is formed by the exposed portion. The micro-needle structure has an inclined sidewall and a through hole surrounded by the inclined sidewall.