Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes

  • Mark Bates
    School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA.
  • Bo Huang
    School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA.
  • Graham T. Dempsey
    School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA.
  • Xiaowei Zhuang
    School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA.

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説明

<jats:p>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 demonstrate 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.</jats:p>

収録刊行物

  • Science

    Science 317 (5845), 1749-1753, 2007-09-21

    American Association for the Advancement of Science (AAAS)

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