FIB Specimen Preparation for Liquid-TEM: A Case Study on Interfacial Mineralization

Abstract Innovations in the liquid-transmission electron microscopy (TEM) field are leading to exciting new possibilities to explore nanoscale dynamic events. Within biological fields, researchers studying biomineralization have leveraged liquid-TEM to probe biomimetic reactions unfolding in real time and to study hydrated samples. However, limited research has been conducted studying interfacial mineralization using this technique. Interfacial mineralization is critical for understanding inorganic-organic interactions, relevant to many biomineralization events, and for studying biomaterials and their interaction with hard tissues. A detailed method for exploring biomaterial interfacial mineralization using nanofabrication with a focused ion beam for in situ liquid-TEM preparation is shared herein. Calcium phosphate (CaP) and titanium (Ti) interfacial mineralization events were observed, where the nucleation and evolution of different CaP particles were noted to form assemblies onto and adjacent to Ti lamellae in situ. With this complementary new method to explore interfacial mineralization mechanisms, this work trailblazes new avenues to study biomineralization.