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Experimental Guidelines to Image Transient Single-Molecule Events Using Graphene Liquid Cell Electron Microscopy.
Huan Wang1,2, Zhun Xu1, Sheng Mao3
1College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, People's Republic of China.
ACS Nano
|October 18, 2022
Summary
Researchers can now visualize molecular interactions in liquids using liquid-phase electron microscopy. This study details methods for successful imaging of synthetic polymers, lipid assemblies, and DNA-enzyme complexes using graphene liquid cells.
Area of Science:
- Materials Science
- Biophysics
- Analytical Chemistry
Background:
- Observing molecular interactions in liquid environments is crucial for understanding biological and chemical processes.
- Current single-molecule imaging methods, including liquid-phase electron microscopy (LPEM), face challenges in achieving sufficient contrast and temporal resolution with safe electron doses.
- Variability in experimental protocols can lead to inconsistent results in LPEM.
Purpose of the Study:
- To present a robust protocol for high-resolution, real-time single-molecule imaging in liquid environments using graphene liquid cells.
- To demonstrate the applicability of this method across diverse molecular systems.
- To provide practical guidance and empirical experience for researchers new to LPEM.
Main Methods:
- Utilized graphene liquid cells for sample containment and electron microscopy.
- Developed detailed procedures for graphene liquid cell fabrication and sample loading.
- Optimized electron microscopy parameters for high contrast and adequate frame rates, focusing on safe electron doses.
- Employed techniques for identifying suitable liquid pockets and target molecules within the cells.
Main Results:
- Successfully imaged dynamic molecular processes in synthetic polymers, lipid assemblies, and DNA-enzyme complexes at nanometer resolution.
- Achieved frame rates sufficient to capture molecular dynamics.
- Demonstrated the reproducibility and effectiveness of the presented experimental tips.
- Provided clear examples of molecular discrimination and process observation.
Conclusions:
- Graphene liquid cells enable high-resolution, time-resolved single-molecule imaging in liquid environments using standard electron microscopes.
- The detailed experimental procedures and empirical experience shared are valuable for researchers aiming to implement LPEM.
- This method significantly advances the ability to study molecular interactions in their native liquid state.
