Journal
SCIENCE
Volume 317, Issue 5845, Pages 1749-1753Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.1146598
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Funding
- NIGMS NIH HHS [R01 GM068518, R01 GM068518-05, GM 068518] Funding Source: Medline
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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.
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