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<p>An <i>in-situ</i> 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.</p>

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FRET-LPTEM for In-situ Imaging of Chemical Systems

  • 期刊论文

  • 发表日期：

    2025年3月11日

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  • Zhun Xu,
  • Deyi Zhang,
  • Tianyu Xiong,
  • Huan Wang

摘要

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.