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Experimental Guidelines to Image Transient Single-Molecule Events Using Graphene Liquid Cell Electron Microscopy.

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In quest of the holy grail to "see" how individual molecules interact in liquid environments, single-molecule imaging methods now include liquid-phase electron microscopy, whose resolution can be nanometers in space… Click to show full abstract

In quest of the holy grail to "see" how individual molecules interact in liquid environments, single-molecule imaging methods now include liquid-phase electron microscopy, whose resolution can be nanometers in space and several frames per second in time using an ordinary electron microscope that is routinely available to many researchers. However, with the current state of the art, protocols that sound similar to those described in the literature lead to outcomes that can differ. The key challenge is to achieve sample contrast under a safe electron dose within a frame rate adequate to capture the molecular process. Here, we present such examples from different systems─synthetic polymer, lipid assembly, DNA-enzyme─in which we have done this using graphene liquid cells. We describe detailed experimental procedures and share empirical experience for conducting successful experiments, starting from fabrication of a graphene liquid cell, to identification of high-quality liquid pockets from desirable shapes and sizes, to effective searching for target sample pockets under electron microscopy, and to discrimination of sample molecules and molecular processes of interest. These experimental tips can assist others who wish to make use of this method.

Keywords: graphene liquid; electron microscopy; microscopy; electron

Journal Title: ACS nano
Year Published: 2022

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