Observation of Liquid-Liquid-Phase Separation and Vesicle Spreading during Supported Bilayer Formation via Liquid-Phase Transmission Electron Microscopy.

Liquid-phase transmission electron microscopy (LP-TEM) enables the real-time visualization of nanoscale dynamics in solution. This technique has been used to study the formation and transformation mechanisms of organic and inorganic nanomaterials. Here, we study the formation of block-copolymer-supported bilayers using LP-TEM. We observe two formation pathways that involve either liquid droplets or vesicles as intermediates toward supported bilayers. Quantitative image analysis methods are used to characterize vesicle spread rates and show the origin of defect formation in supported bilayers. Our results suggest that bilayer assembly methods that proceed via liquid droplet intermediates should be beneficial for forming pristine supported bilayers. Furthermore, supported bilayers inside the liquid cells may be used to image membrane interactions with proteins and nanoparticles in the future.