In Situ Visualization of Ferritin Biomineralization via Graphene Liquid Cell-Transmission Electron Microscopy

Ferritin biomineralization is essential to regulate the toxic Fe²⁺ iron ions in the human body. Unravelling the mechanism of biomineralization in ferritin facilitates our understanding of the causes underlying many iron disorder-related diseases. Until now, no report of in situ visualization of ferritin biomineralization events at nanoscale exists due to the requirement for high-resolution imaging of nanometer-sized ferritin proteins in their hydrated states. Herein, for the first time, we show that the biomineralization processes within individual ferritin proteins can be visualized by means of graphene liquid cell-transmission electron microscopy (GLC-TEM). The increase in the ratio of Fe³⁺/Fe²⁺ ions over time monitored via electron energy loss spectroscopy (EELS) reveals the change in oxidation state of iron oxide phases with time. This study lays a foundation for future investigations on iron regulation mechanisms in healthy and dysfunctional ferritins.