Mollusk biomineralization is an elaborate process in which cells, organic macromolecules, and calcium carbonate crystals are actively involved.Macromolecules (mainly are proteins and polysaccharide) act as a key role in regulating and limiting the size, orientation, polymorph and texture of inorganic phase. In this work, we focused on the inter-prismatic matrix of mollusk shell combining scanning electron microscopy (SEM), X-ray diffraction (XRD) and transmission electron microscopy (TEM) analytical techniques with CaCO 3 recrystallization experiment to characterize its structure and effects on crystal formation. Our results show that the inter-prismatic matrix is not a sort of pure polymer, calcite nano-crystals are also located inside the inter-prismatic matrix. Interestingly, it seems that these nanocrystals have a preferred orientation, which means the inter-prismatic matrix do impose effect on the crystal formation. In vitro re-crystallization experiment using partially dissolved prismatic fragment as template indicates that the (104) faces of CaCO 3 micro-crystals are closely associated with the walls of inter-prismatic matrix. Furthermore, a possible growth mechanism of mollusk shell prismatic layer was proposed.Key Words: Biomineralization, Mollusk shell, Inter-prismatic matrix, Calcite, Dissolution Citation: D. Yang, et al. Inter-prismatic matrix structure characterization of mollusk shell and its effect on crystal formation. Nano Biomed. Eng.