Three kinds of natural starch, corn starch, waxy corn starch and cassava starch, were used as raw materials to prepare starch-based microspheres by dropping method after high temperature gelatinization. The effects of starch type, starch concentration and needle aperture on microsphere formation were investigated. It was found that the particle size of the microspheres increased with the increase of starch concentration and needle aperture, and the corn starch-based microspheres showed the largest particle size at the same starch concentration and needle aperture. The starch-based microspheres have uniform morphology, and its yield can reach more than 90%. The particle size of the microspheres was 1.0~1.5 mm, and they had different degrees of the hollow structure. Compared with the natural starch, the gelatinization exothermic peak of starch-based microspheres basically disappeared, the crystal type changed and the crystallinity decreased significantly. The α-amylase enzymolysis rate of corn starch microspheres was higher than that of corn starch at 24 h, and the release rate of the loaded Methylene blue corn starch based microspheres could reach more than 59.28% after heating in aqueous solution at 30°C ~90°C for 1 h, indicating that they were beneficial to load various hydrophilic substances.