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Fluorescence microscopy

Strategic blinking

Nature Chemistry volume 6, pages 663–664 (2014)Cite this article

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For decades chemists have focused on increasing the brightness of fluorophores. In super-resolution microscopy, however, fluorophores that preferentially exist in a non-fluorescent state, but occasionally re-arrange into a fluorescent form, can give better results.

Super-resolution microscopy — the imaging of objects with a resolution below the diffraction barrier of light — has enabled numerous important discoveries in biology1. One approach is single-molecule localization microscopy (SLM), in which individual fluorophores are separated by stochastically switching them between fluorescent 'on' and non-fluorescent 'off' states. At any given time, only a sparse subset of fluorophores are in the on state, and therefore detected and localized. Repeating this process thousands of times enables reconstruction of an image with a resolution below the diffraction barrier1.

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Figure 1: Construction of super-resolution images using the hydroxymethyl silicon-rhodamine (HMSiR) dye.

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Authors and Affiliations

  1. Gražvydas Lukinavičius and Kai Johnsson are in the École Polytechnique Fédérale de Lausanne (EPFL), Institute of Chemical Sciences and Engineering, NCCR Chemical Biology, 1015 Lausanne, Switzerland,

    Gražvydas Lukinavičius & Kai Johnsson

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  1. Gražvydas Lukinavičius
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  2. Kai Johnsson
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Correspondence to Kai Johnsson.

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Lukinavičius, G., Johnsson, K. Strategic blinking. Nature Chem 6, 663–664 (2014). https://doi.org/10.1038/nchem.2020

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