Localization of intracrystalline organic macromolecules in mollusk shells

As a crucial first step for understanding the organic–inorganic interaction in biomineralization of mollscan shells, localization of the intracrystalline organic macromolecules in biogenic calcium carbonate crystals of the nacreous, prismatic (Pinctada fucata) and foliated (Cellana toreuma) microstructures were investigated using Fresnel contrast analysis in a transmission electron microscope. Spherular Fresnel contrasts in the crystals correspond to organic substances, which was confirmed by the detection of 1s→π⁎ (CC) transition peak at 284 eV in electron energy loss spectroscopy. Nano-sized (5–10 nm) spherules in the aragonite tablets constituting the nacreous layer of P. fucata specifically concentrate in the vicinity of the interlamellar membrane between the aragonite tablets. The dominant sizes of the organic macromolecules extracted by dissolving the aragonite tablets in the nacreous layer of P. fucata were estimated using the gel-filtration analysis to be roughly 10 and 4 nm, which dimensionally corresponds to the sizes observed by Fresnel contrast imaging in the tablets. These results will serve for understanding the functions of intracrystalline organic macromolecules in mollusk shells.

► The intracrystalline organic macromolecules were visualized using Fresnel contrasts.
► EELS analysis showed Spherular Fresnel contrasts contained organic macromolecules.
► The size of these contrasts corresponded to that of extracted organic substances.
► Localization of the organic macromolecules suggests the hypothesis of crystal formation process.