Layer-by-layer (LbL) assembly is a most commonly used method to prepare various microcapsules based on the electrostatic interactions, hydrogen bonding, covalent bonding, and so on. Among these interactions, Schiff base bond formed in covalent assembly not only has an advantage in stability, but also enables the assembled microcapsules with autofluorescence and pH sensitivity. In this Article, we will mainly describe the construction of biomimetic microcapsules through Schiff base mediated LbL assembly. The structures and properties of the assembled microcapsules are introduced and their applications as drug carriers are highlighted.