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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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
Subjects:
Online Access:https://doi.org/10.1002/advs.202408280
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author Seong Ho Kim
Adam B. Yasunaga
Hongyuan Zhang
Kevin D. Whitley
Isaac T. S. Li
author_facet Seong Ho Kim
Adam B. Yasunaga
Hongyuan Zhang
Kevin D. Whitley
Isaac T. S. Li
author_sort Seong Ho Kim
collection DOAJ
description 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.
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spelling doaj-art-92092d2d53b44482bd2ba6c5ce8a50a72025-08-20T03:15:34ZengWileyAdvanced Science2198-38442025-07-011228n/an/a10.1002/advs.202408280Quantitative Super‐Resolution Imaging of Molecular TensionSeong Ho Kim0Adam B. Yasunaga1Hongyuan Zhang2Kevin D. Whitley3Isaac T. S. Li4Department of Chemistry University of British Columbia Kelowna BC V1V 1V7 CanadaDepartment of Chemistry University of British Columbia Kelowna BC V1V 1V7 CanadaDepartment of Chemistry University of British Columbia Kelowna BC V1V 1V7 CanadaCentre for Bacterial Cell Biology Biosciences Institute Newcastle University Newcastle upon Tyne NE1 7RU UKDepartment of Chemistry University of British Columbia Kelowna BC V1V 1V7 CanadaAbstract 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.https://doi.org/10.1002/advs.202408280DNA‐PAINTfunctional super‐resolution imagingmolecular beaconmolecular tension sensor
spellingShingle Seong Ho Kim
Adam B. Yasunaga
Hongyuan Zhang
Kevin D. Whitley
Isaac T. S. Li
Quantitative Super‐Resolution Imaging of Molecular Tension
Advanced Science
DNA‐PAINT
functional super‐resolution imaging
molecular beacon
molecular tension sensor
title Quantitative Super‐Resolution Imaging of Molecular Tension
title_full Quantitative Super‐Resolution Imaging of Molecular Tension
title_fullStr Quantitative Super‐Resolution Imaging of Molecular Tension
title_full_unstemmed Quantitative Super‐Resolution Imaging of Molecular Tension
title_short Quantitative Super‐Resolution Imaging of Molecular Tension
title_sort quantitative super resolution imaging of molecular tension
topic DNA‐PAINT
functional super‐resolution imaging
molecular beacon
molecular tension sensor
url https://doi.org/10.1002/advs.202408280
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AT kevindwhitley quantitativesuperresolutionimagingofmoleculartension
AT isaactsli quantitativesuperresolutionimagingofmoleculartension