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Super-resolution Imaging of the Bacterial Division Machinery
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Quantitative Super-Resolution Imaging of Molecular Tension.

Seong Ho Kim1,2, Adam B Yasunaga1, Hongyuan Zhang1

  • 1Department of Chemistry, University of British Columbia, Kelowna, BC, V1V 1V7, Canada.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|April 17, 2025
PubMed
Summary

This study introduces quantitative tension points accumulation for imaging in nanoscale topography (qtPAINT), a new method to precisely measure molecular tension forces in live cells using DNA probes with super-resolution imaging.

Keywords:
DNA‐PAINTfunctional super‐resolution imagingmolecular beaconmolecular tension sensor

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Area of Science:

  • Biophysics
  • Molecular Biology
  • Nanotechnology

Background:

  • DNA-based molecular tension probes enable super-resolution localization of mechanical events in cells.
  • Quantifying the magnitude of these forces at super-resolution has remained a significant challenge.

Purpose of the Study:

  • To introduce a novel super-resolution imaging strategy, qtPAINT, for accurately measuring molecular tension magnitude.
  • To enable quantitative analysis of force magnitudes acting on individual tension probes within live cells.

Main Methods:

  • Developed qtPAINT by integrating a molecular-beacon PAINT imager with a hairpin molecular tension probe.
  • Leveraged force-dependent dissociation kinetics of DNA oligonucleotides to encode tension based on binding kinetics.
  • Enabled quantitative analysis of binding kinetics for force reconstruction.

Main Results:

  • Achieved super-resolution accuracy in imaging molecular tension magnitude.
  • Demonstrated a force quantification range of 9–30 pN.
  • Maintained spatial resolution of 30–120 nm and temporal resolution on the order of a minute.

Conclusions:

  • qtPAINT provides a powerful tool for super-resolution imaging of molecular tension magnitude.
  • The method enhances the study of dynamic cellular mechanical processes.
  • Offers a significant advancement in quantitative force microscopy.