Multicolor super-resolution imaging with photo-switchable fluorescent probes.
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.
Bates
Mark
M
School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA.
Huang
Bo
B
Dempsey
Graham T
GT
Zhuang
Xiaowei
X
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D022163:Clathrin-Coated Vesicles; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D005456:Fluorescent Dyes; D008856:Microscopy, Fluorescence / Q000379:methods; D008870:Microtubules; D036103:Nanotechnology
D015342:DNA Probes; D005456:Fluorescent Dyes; D004247:DNA
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