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

Niels Radmacher; Oleksii Nevskyi; José Ignacio Gallea; Jan Christoph Thiele; Ingo Gregor; Jörg Enderlein

doi:10.1117/12.3004932

13 March 2024 Doubling the resolution of single-molecule localization microscopy with image scanning microscopy

Niels Radmacher, Oleksii Nevskyi, José Ignacio Gallea, Jan Christoph Thiele, Ingo Gregor, Jörg Enderlein

Proceedings Volume PC12849, Single Molecule Spectroscopy and Superresolution Imaging XVII; PC128490I (2024) https://doi.org/10.1117/12.3004932
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

In recent years, Image Scanning Microscopy (ISM) has emerged as a powerful technique for achieving super-resolution bio-imaging across various applications. Particularly noteworthy is the implementation of a single-photon detector array, enabling the utilization of Lifetime Image Scanning Microscopy, which has proven to be highly effective. In our study, we present a novel approach that combines ISM with direct Stochastic Optical Reconstruction Microscopy (dSTORM), resulting in a doubling of the localization precision in Single Molecule Localization Microscopy (SMLM). Additionally, we capitalize on the available lifetime information provided by ISM, allowing for multilabel fluorescence measurements without the detrimental effects of chromatic aberration, even at resolutions significantly surpassing the diffraction limit. Moreover, we introduce a freely available add-on to previously employed open-source tools for single particle tracking and localization, enhancing the accessibility and utility of our methodology. This add-on serves as a valuable resource for the research community, facilitating the adoption and further advancement of the combined ISM and dSTORM technique.

Conference Presentation

(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.

Citation Download Citation

Niels Radmacher, Oleksii Nevskyi, José Ignacio Gallea, Jan Christoph Thiele, Ingo Gregor, and Jörg Enderlein "Doubling the resolution of single-molecule localization microscopy with image scanning microscopy", Proc. SPIE PC12849, Single Molecule Spectroscopy and Superresolution Imaging XVII, PC128490I (13 March 2024); https://doi.org/10.1117/12.3004932

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KEYWORDS

Image resolution

Fluorescence

Microscopes

Confocal microscopy

Detector arrays

Multiplexing

Open source software

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