Quantitative Super‐Resolution Imaging of Molecular Tension

Abstract DNA‐based molecular tension probes have revolutionized the localization of mechanical events in live cells with super‐resolution. However, imaging the magnitude of these forces at super‐resolution has been challenging. Here, qtPAINT (quantitative tension points accumulation for imaging in n...

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Bibliographic Details
Main Authors: Seong Ho Kim, Adam B. Yasunaga, Hongyuan Zhang, Kevin D. Whitley, Isaac T. S. Li
Format: Article
Language:English
Published: Wiley 2025-07-01
Series:Advanced Science
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Online Access:https://doi.org/10.1002/advs.202408280
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Summary:Abstract DNA‐based molecular tension probes have revolutionized the localization of mechanical events in live cells with super‐resolution. However, imaging the magnitude of these forces at super‐resolution has been challenging. Here, qtPAINT (quantitative tension points accumulation for imaging in nanoscale topography) is introduced as a strategy to image the magnitude of molecular tension with super‐resolution accuracy. By leveraging the force‐dependent dissociation kinetics of short DNA oligonucleotides on their complementary strands, tension is encoded on individual molecules through their binding kinetics. This method allowed for a quantitative analysis of these kinetics, providing a detailed reconstruction of the force magnitudes acting on each tension probe. The technique integrates a molecular‐beacon PAINT imager with a hairpin molecular tension probe, achieving a force quantification range of 9–30 pN and maintaining a spatial resolution of 30–120 nm in low and high‐density regions. Additionally, qtPAINT offers a temporal resolution on the order of a minute, enhancing its applicability for studying dynamic cellular processes.
ISSN:2198-3844