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Quantitative super-resolution imaging of molecular tension

Lookup NU author(s): Dr Kevin WhitleyORCiD

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

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.

Publication metadata

Author(s): Kim SH, Yasunaga AB, Zhang H, Whitley KD, Li ITS

Publication type: Article

Publication status: Published

Journal: Advanced Science

Year: 2025

Volume: 12

Issue: 28

Print publication date: 24/07/2025

Online publication date: 17/04/2025

Acceptance date: 07/03/2025

Date deposited: 16/05/2025

ISSN (electronic): 2198-3844

Publisher: Wiley-VCH Verlag GmbH & Co. KGaA

URL: https://doi.org/10.1002/advs.202408280

DOI: 10.1002/advs.202408280

Data Access Statement: The data that support the ?ndings of this study are available from the corresponding author upon reasonable request.

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