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Related Experiment Video

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Label-Retention Expansion Microscopy LR-ExM Enables Super-Resolution Imaging and High-Efficiency Labeling
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Prelabeling Expansion Single-Molecule Localization Microscopy with Minimal Linkage Error.

Minsu Kang1,2, Jooyong Lee3, Sangyoon Ko1,2

  • 1Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science (IBS), Seoul, 02841, Republic of Korea.

Chembiochem : a European Journal of Chemical Biology
|December 16, 2020
PubMed
Summary

This study introduces a new trifunctional streptavidin method to improve expansion microscopy combined with single-molecule localization microscopy (ExSMLM). This technique achieves high-resolution molecular imaging with minimal linkage error.

Keywords:
expansion microscopyfluorescencesingle-molecule localization microscopysuper-resolution microscopy

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Area of Science:

  • Biochemistry
  • Microscopy
  • Molecular Biology

Background:

  • Expansion microscopy combined with single-molecule localization microscopy (ExSMLM) offers potential for molecular resolution imaging.
  • Current ExSMLM methods face challenges including fluorophore/protein loss and increased linkage error.

Purpose of the Study:

  • To develop an improved ExSMLM technique addressing current limitations.
  • To enhance labeling efficiency and reduce linkage error in molecular resolution imaging.

Main Methods:

  • Introduction of a trifunctional streptavidin system.
  • Linking target molecules, fluorophores, and gel matrix via a biotinylizable peptide tag.
  • Application to imaging vimentin filaments.

Main Results:

  • Achieved high labeling efficiency in ExSMLM.
  • Demonstrated a minimal linkage error of approximately 5 nm.
  • Successfully imaged vimentin filaments with enhanced resolution.

Conclusions:

  • The novel trifunctional streptavidin approach significantly improves ExSMLM.
  • This method provides a practical and simple way to achieve molecular resolution in fluorescence imaging.