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Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes

Mark Bates, Bo Huang, Graham T. Dempsey and Xiaowei Zhuang
Science
New Series, Vol. 317, No. 5845 (Sep. 21, 2007), pp. 1749-1753
Stable URL: http://www.jstor.org/stable/20048431
Page Count: 5
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Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes
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Abstract

Recent advances in far-field optical nanoscopy have enabled fluorescence imaging with a spatial resolution of 20 to 50 nanometers. Multicolor super-resolution imaging, however, remains a challenging task. Here, we introduce a family of photo-switchable fluorescent probes and demostrate multicolor stochastic optical reconstruction microscopy (STORM). Each probe consists of a photo-switchable "reporter" fluorophore that can be cycled between fluorescent and dark states, and an "activator" that facilitates photo-activation of the reporter. Combinatorial pairing of reporters and activators allows the creation of probes with many distinct colors. Iterative, color-specific activation of sparse subsets of these probes allows their localization with nanometer accuracy, enabling the construction of a super-resolution STORM image. Using this approach, we demonstrate multicolor imaging of DNA model samples and mammalian cells with 20- to 30-nanometer resolution. This technique will facilitate direct visualization of molecular interactions at the nanometer scale.

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