Sanders, EW, Carr, AR, Bruggeman, E et al. (10 more authors) (2022) resPAINT: Accelerating volumetric super‐resolution localisation microscopy by active control of probe emission. Angewandte Chemie International Edition, 61 (42). e202206919. ISSN 1433-7851
Abstract
Points for accumulation in nanoscale topography (PAINT) allows practically unlimited measurements in localisation microscopy but is limited by background fluorescence at high probe concentrations, especially in volumetric imaging. We present reservoir-PAINT (resPAINT), which combines PAINT and active control of probe photophysics. In resPAINT, an activatable probe ‘reservoir’ accumulates on target, enabling a 50-fold increase in localisation rate versus conventional PAINT, without compromising contrast. By combining resPAINT with large depth-of-field microscopy, we demonstrate super-resolution imaging of entire cell surfaces. We generalise the approach by implementing various switching strategies and 3D imaging techniques. Finally, we use resPAINT with a Fab to image membrane proteins, extending the operating regime of PAINT to include a wider range of biological interactions.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
Keywords: | PAINT; super-resolution microscopy; Localisation microscopy; Biophysics; Single-molecule imaging |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Physics and Astronomy (Leeds) > Molecular & Nanoscale Physics |
Funding Information: | Funder Grant number Academy of Medical Sciences SBF006\1138 |
Depositing User: | Symplectic Publications |
Date Deposited: | 11 Aug 2022 15:43 |
Last Modified: | 29 Nov 2022 02:21 |
Status: | Published |
Publisher: | Wiley |
Identification Number: | 10.1002/anie.202206919 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:189860 |