Repository logo
 

resPAINT: Accelerating Volumetric Super-Resolution Localisation Microscopy by Active Control of Probe Emission.

Published version
Peer-reviewed

Change log

Authors

Carr, Alexander R 
Bruggeman, Ezra 
Körbel, Markus 
Benaissa, Sarah I 

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.

Description

Keywords

Biophysics, Localisation Microscopy, PAINT, Single-Molecule Imaging, Super-Resolution Microscopy

Journal Title

Angew Chem Weinheim Bergstr Ger

Conference Name

Journal ISSN

0044-8249
1521-3757

Volume Title

Publisher

Wiley
Sponsorship
Engineering and Physical Sciences Research Council (EP/M003663/1)
Wellcome Trust (206291/Z/17/Z)