FRET-LPTEM for In-situ Imaging of Chemical Systems

doi:10.1007/s40242-025-4241-7

Chemical Research in Chinese Universities ›› 2025, Vol. 41 ›› Issue (2): 319-325.doi: 10.1007/s40242-025-4241-7

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FRET-LPTEM for In-situ Imaging of Chemical Systems

XU Zhun, ZHANG Deyi, XIONG Tianyu, WANG Huan

Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Key Laboratory of Polymer Chemistry and Physics, National Biomedical Imaging Center, Peking University, Beijing 100871, P. R. China

Received:2024-12-14 Accepted:2025-01-25 Online:2025-04-01 Published:2025-03-31
Contact: WANG Huan,wanghuan_ccme@pku.edu.cn E-mail:wanghuan_ccme@pku.edu.cn
Supported by:
This work was supported by the Natural Science Foundation of Beijing Municipality, China (No. Z240010) and the National Natural Science Foundation of China (Nos. 22174006, T2495223). We thank Prof. SHAO Yuanhua for supporting the CO2-laser-based puller (P-2000, Sutter Instrument Co.) in making nanopipette liquid cells.

Abstract

Abstract: An in-situ double-tilt holder has been made to integrate laser illumination and fluorescence-based spectroscopic analysis for conducting liquid-phase electron microscopy (LP-TEM) experiments using ordinary TEM. The setup differs from the existing geometry. Laser illumination and the collection of fluorescence signals were achieved using a single optical fiber, with efficiency optimized by adjusting the fiber position and grid tilt angle. Fluorescence emission of common organic dyes, propidium iodide (PI) and cyanine dyes, and Förster resonance energy transfer (FRET) signals of a FRET pair, Cy3/Cy5, were obtained from three types of liquid cells, including carbon film, graphene, and nanopipette liquid cells. The successful application of FRET-LPTEM enables LP-TEM experiments to be equipped with controlled light-triggering capability, detection of fluorogenic small molecules during chemical reactions, and the standard FRET experiments for macromolecules being conducted with LP-TEM. FRET-LPTEM presents opportunities for unraveling pathways underpinning the synthesis and assembly of optically active organic and biological materials.

Key words: In-situ double-tilt holder, Optical fiber, Light-coupling experiment, Förster resonance energy transfer (FRET), Liquidphase electron microscopy (LP-TEM)

XU Zhun, ZHANG Deyi, XIONG Tianyu, WANG Huan. FRET-LPTEM for In-situ Imaging of Chemical Systems[J]. Chemical Research in Chinese Universities, 2025, 41(2): 319-325.

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FRET-LPTEM for In-situ Imaging of Chemical Systems

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