Experimental Guidelines to Image Transient Single-Molecule Events Using Graphene Liquid Cell Electron Microscopy
Source abstract
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.
Source: openalex_inverted_index · Quality: baseline
Research context
OpenAlex topics: Advanced Electron Microscopy Techniques and Applications · Advanced Fluorescence Microscopy Techniques · Molecular Junctions and Nanostructures
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, People’s Republic of China
Beijing National Laboratory for Molecular Sciences, Center for Spectroscopy, Beijing Key Laboratory of Polymer Chemistry & Physics of Ministry of Education, Center for Soft Matter Science and Engineering, National Biomedical Imaging Center, Peking University, Beijing 100871, People’s Republic of China
Department of Mechanics and Engineering Sciences, College of Engineering, Peking University, Beijing 100871, People’s Republic of China
Center for Soft and Living Matter, Institute for Basic Science, Ulsan, Korea, 44919
Department of Chemistry and Physics, Ulsan National Institute of Science and Technology, Ulsan, Korea 44919
Funding
Institute for Basic Science · National Natural Science Foundation of China · Beijing National Laboratory for Molecular Sciences · Center for Spectroscopic Analysis of College of Chemistry and Molecular Engineering, Peking University
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