Updated in 1/22/2009 3:31:46 PM      Viewed: 552 times      (Journal Article)
Science (New York, N.Y.) 317 (5845): 1749-53 (2007)

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

Mark Bates , Bo Huang , Graham T Dempsey , Xiaowei Zhuang
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.
DOI: 10.1126/science.1146598      ISSN: 0036-8075