Doubling the resolution of fluorescence-lifetime single-molecule localization microscopy with image scanning microscopy

In this study, we integrate a single-photon detector array into a confocal laser scanning microscope, enabling the combination of fluorescence-lifetime single-molecule localization microscopy with image scanning microscopy. This unique combination delivers a twofold improvement in lateral localizati...

Full description

Saved in:
Bibliographic Details
Published in:Nature photonics 2024-10, Vol.18 (10), p.1059-1066
Main Authors: Radmacher, Niels, Nevskyi, Oleksii, Gallea, José Ignacio, Thiele, Jan Christoph, Gregor, Ingo, Rizzoli, Silvio O., Enderlein, Jörg
Format: Article
Language:English
Subjects:
639/624/1107/328/2238
639/925/930/328/2238
Accuracy
Applied and Technical Physics
Approximation
Biophysics
Efficiency
Fluorescence
Image enhancement
Image processing
Image reconstruction
Laser applications
Laser arrays
Localization
Microscopy
Multiplexing
Photonics
Photons
Physics
Physics and Astronomy
Quantum Physics
Scanning microscopy
Sensors
Stochasticity
Topography
Citations: Items that this one cites
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:In this study, we integrate a single-photon detector array into a confocal laser scanning microscope, enabling the combination of fluorescence-lifetime single-molecule localization microscopy with image scanning microscopy. This unique combination delivers a twofold improvement in lateral localization accuracy for single-molecule localization microscopy (SMLM) and maintains its simplicity. Moreover, the addition of lifetime information from our confocal laser scanning microscope eliminates chromatic aberration, particularly crucial for achieving few-nanometre resolution in SMLM. Our approach, named fluorescence-lifetime image scanning microscopy SMLM, is demonstrated through direct stochastic optical reconstruction microscopy and DNA point accumulation for imaging in nanoscale topography experiments on fluorescently labelled cells, showcasing both resolution enhancement and fluorescence-lifetime multiplexing capabilities. The integration of a single-photon detector array and imaging scanning microscopy in a confocal scanning microscope enables doubling the resolution of single-molecule localization microscopy.
ISSN:1749-4885
1749-4893
DOI:10.1038/s41566-024-01481-4