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Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...

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Application of Monolayer Graphene to Cryo-Electron Microscopy Grids for High-resolution Structure Determination
07:57

Application of Monolayer Graphene to Cryo-Electron Microscopy Grids for High-resolution Structure Determination

Published on: November 10, 2023

Exploring nanographene for single molecule imaging at cryogenic temperatures.

Yutong Wang1, Qiqi Yang2, Xiaomin Liu2

  • 1Department of Imaging Physics, Delft University of Technology, Delft, The Netherlands.

The Journal of Chemical Physics
|June 24, 2026
PubMed
Summary
This summary is machine-generated.

Single nanographene fluorophores (DBOV-azide) exhibit temperature-dependent blinking and enhanced photon output at cryogenic temperatures, crucial for advanced microscopy techniques.

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Microscopic Visualization of Porous Nanographenes Synthesized through a Combination of Solution and On-Surface Chemistry

Published on: March 4, 2021

Area of Science:

  • Cryo-electron microscopy
  • Single-molecule imaging
  • Fluorescence microscopy

Background:

  • Cryogenic temperature imaging enhances photon budgets by reducing photobleaching and non-radiative decay.
  • Correlative cryo-fluorescence and electron microscopy requires fluorophores with stable low-temperature blinking properties.
  • Nanographenes offer self-blinking fluorescence but their low-temperature behavior is unquantified.

Purpose of the Study:

  • To characterize the low-temperature blinking intermittency and photon output of dibenzo[hi,st]ovalene (DBOV-azide) nanographene fluorophores.
  • To evaluate DBOV-azide as a potential fluorophore for cryogenic single-molecule imaging.
  • To understand the influence of temperature and excitation irradiance on nanographene photophysics.

Main Methods:

  • Single-molecule fluorescence spectroscopy at temperatures ranging from 91 K to 293 K.
  • Wide-field imaging with generalized likelihood ratio test for on/off state extraction.
  • Analysis of on-time, off-time distributions, and photon counts under varying excitation irradiances (1.2-5.9 kW cm-2).

Main Results:

  • DBOV-azide exhibits blinking across all tested temperatures, with longer on-times at 91 K compared to room temperature.
  • On- and off-time distributions follow power laws, independent of temperature and irradiance for short timescales (<1.5 s).
  • Photon output per on-event increases significantly at cryogenic temperatures, with a ~10x decrease in the on/off ratio from 91 K to 293 K.

Conclusions:

  • DBOV-azide demonstrates promising photophysical properties for cryogenic single-molecule imaging, despite temperature-dependent blinking characteristics.
  • The enhanced photon output at low temperatures makes nanographenes valuable for applications requiring high signal-to-noise ratios.
  • Further optimization of nanographene structures may improve duty cycles for even broader cryogenic microscopy applications.