Exploring Biomineralization Processes Using In Situ Liquid Transmission Electron Microscopy: A Review

Liquid transmission electron microscopy (TEM) is a newly established technique broadly used to study reactions in situ. Since its emergence, complex and multifaceted biomineralization processes have been revealed with real-time resolution, where classical and non-classical mineralization pathways have been dynamically observed primarily for Ca and Fe-based mineral systems in situ. For years, classical crystallization pathways have dominated theories on biomineralization progression despite observations of non-traditional routes involving precursor phases using traditional- and cryo-TEM. The new dynamic lens provided by liquid TEM is a key correlate to techniques limited to time-stamped, static observations - helping shift paradigms in biomineralization toward non-classical theories with dynamic mechanistic visualization. Liquid TEM provides new insights into fundamental biomineralization processes and essential physiological and pathological processes for a wide range of organisms. This review critically reviews a summary of recent in situ liquid TEM research related to the biomineralization field. Key liquid TEM preparation and imaging parameters are provided as a foundation for researchers while technical challenges are discussed. In future, the expansion of liquid TEM research in the biomineralization field will lead to transformative discoveries, providing complementary dynamic insights into biological systems.