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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
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.
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.
