Increasing speed and brightness in super-resolution microscopy with renewable fluorophores

Marina S. Dietz; Soohyen Jang; Johanna Rahm; Laurell Kessler; Ashwin Balakrishnan; Nina S. Deußner-Helfmann; Yunqing Li; Kaarjel Narayanasamy; Hans-Dieter Barth; Mike Heilemann

doi:10.1117/12.3009172

12 March 2024 Increasing speed and brightness in super-resolution microscopy with renewable fluorophores

Marina S. Dietz, Soohyen Jang, Johanna Rahm, Laurell Kessler, Ashwin Balakrishnan, Nina S. Deußner-Helfmann, Yunqing Li, Kaarjel Narayanasamy, Hans-Dieter Barth, Mike Heilemann

Author Affiliations +

Proceedings Volume 12849, Single Molecule Spectroscopy and Superresolution Imaging XVII; 1284903 (2024) https://doi.org/10.1117/12.3009172
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Fluorophore labels that transiently and repetitively bind to a target (“exchangeable” or “renewable” labels) lead to a continuous renewal of the fluorescence signal. This dynamic labeling approach minimizes photobleaching and was beneficially exploited in various super-resolution microscopy methods. Here, we report two new developments using exchangeable fluorophores: first, we report fast and long-time live-cell super-resolution microscopy using a weak-affinity protein label and a neural network. Second, we report a novel design for exchangeable DNA labels that show higher brightness and lower background. Together, these two developments further increase the application range for exchangeable fluorophore labels in super-resolution fluorescence microscopy.

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Marina S. Dietz, Soohyen Jang, Johanna Rahm, Laurell Kessler, Ashwin Balakrishnan, Nina S. Deußner-Helfmann, Yunqing Li, Kaarjel Narayanasamy, Hans-Dieter Barth, and Mike Heilemann "Increasing speed and brightness in super-resolution microscopy with renewable fluorophores", Proc. SPIE 12849, Single Molecule Spectroscopy and Superresolution Imaging XVII, 1284903 (12 March 2024); https://doi.org/10.1117/12.3009172

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