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Science. 2007 Sep 21;317(5845):1749-53. Epub 2007 Aug 16.

Multicolor super-resolution imaging with photo-switchable fluorescent probes.

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  • 1School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA.

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 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.

PMID:
17702910
[PubMed - indexed for MEDLINE]
PMCID:
PMC2633025
Free PMC Article
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