Fluorescence nanoscopy by ground-state depletion and single-molecule return

By J. Fölling, Mariano L. Bossi, H. Bock, R. Medda, C. Wurm, B. Hein, S. Jakobs, Christian Eggeling, Stefan W. Hell. Nature Methods, 2008

759 citations recorded by the source corpus.

We introduce far-field fluorescence nanoscopy with ordinary fluorophores based on switching the majority of them to a metastable dark state, such as the triplet, and calculating the position of those left or those that spontaneously returned to the ground state. Continuous widefield illumination by a single laser and a continuously operating camera yielded dual-color images of rhodamine- and fluorescent protein–labeled (living) samples, proving a simple yet powerful super-resolution approach.

Subfields: Biomedical Engineering, Biophysics, Structural Biology

View Paper: https://www.semanticscholar.org/paper/10451477729b499350142ba2fc94c5668ed1d021

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Citations in the corpus

Image Resolution in Optical Nanoscopy (2013, background): “These clusters may be due to, for example, anomalous fluorescent (5) molecules that are active during a large part of the total measurement time.”
Empirical optimal transport on countable metric spaces: Distributional limits and statistical applications (2017, background): “Single Marker Switching (SMS) Microscopy (Betzig et al., 2006; Rust et al., 2006; Egner et al., 2007; Heilemann et al., 2008; Fölling et al., 2008) is a living cell fluorescence microscopy technique in which fluorescent markers which are tagged to a protein structure in the probe are stochastically”
Direct stochastic optical reconstruction microscopy with standard fluorescent probes (2011, methodology): “, ground-state depletion microscopy followed by individual molecule retur”
Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging (2011, positive): “Like Alexa 647, a variety of commonly used organic dyes spanning a broad spectrum of colors can transition between fluorescent and dark states and have been used for super-resolution imaging 8,10,11,14–16 .”
Frontiers in structured illumination microscopy (2016, background): “Notable examples include photoactivated localization microscopy (PALM) [7], fluorescence PALM [8], stochastic optical reconstruction microscopy (STORM) [9], and direct STORM [10], with the latter also known as ground state depletion microscopy followed by individual molecule return [11].”