Exchangeable fluorophore labels in super-resolution fluorescence microscopy

Mike Heilemann

doi:10.1117/12.2612522

2 March 2022 Exchangeable fluorophore labels in super-resolution fluorescence microscopy

Mike Heilemann

Author Affiliations +

Proceedings Volume PC11967, Single Molecule Spectroscopy and Superresolution Imaging XV; PC119670C (2022) https://doi.org/10.1117/12.2612522
Event: SPIE BiOS, 2022, San Francisco, California, United States

Abstract

We present fluorophore labels that transiently and repetitively bind to their targets as probes for various types of super-resolution fluorescence microscopy. Such labels show a weak (~10 µM – 100 nM) affinity to a target and are kept in an imaging buffer that constitutes a reservoir with a high concentration of intact probes, enabling repetitive binding to the same target (we refer to these labels as “exchangeable labels”). This dynamic labeling approach minimizes photobleaching and yields a constant fluorescence signal over time, which has been beneficially exploited in SMLM [1-4], STED [5, 6], and super-resolution optical fluctuation imaging (SOFI) [7]. Multi-color, 3D, and live cell imaging, as well as imaging of large fields of view, is facilitated [4]. We further present the implementation of neural networks for multi-emitter localization to achieve multi-color SMLM with short acquisition times of one minute [8].

Conference Presentation

Citation Download Citation

Mike Heilemann "Exchangeable fluorophore labels in super-resolution fluorescence microscopy", Proc. SPIE PC11967, Single Molecule Spectroscopy and Superresolution Imaging XV, PC119670C (2 March 2022); https://doi.org/10.1117/12.2612522

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