Volume 135, Issue 7 e202217028
Zuschrift

Super-Resolution Tension PAINT Imaging with a Molecular Beacon

Dr. Seong Ho Kim

Dr. Seong Ho Kim

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

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Prof. Dr. Isaac T. S. Li

Corresponding Author

Prof. Dr. Isaac T. S. Li

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

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First published: 19 December 2022

Abstract

DNA-PAINT enabled super-resolution imaging through the transient binding of fluorescently-labelled single-stranded DNA (ssDNA) imagers to target ssDNA. However, its performance is constrained by imager background fluorescence, resulting in relatively long image acquisition and potential artifacts. We designed a molecular beacon (MB) as the PAINT imager. Unbound MB in solution reduces the background fluorescence due to its natively quenched state. They are fluorogenic upon binding to target DNA to create individual fluorescence events. We demonstrate that MB-PAINT provides localization precision similar to traditional linear imager DNA-PAINT. We also show that MB-PAINT is ideally suited for fast super-resolution imaging of molecular tension probes in living cells, eliminating the potential of artifacts from free-diffusing imagers in traditional DNA-PAINT at the cell-substrate interface.

Data Availability Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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