DOI: 10.1126/science.1146598. Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes

Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes

10.1126/science.1146598

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

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.

Bibliography

Bates, M., Huang, B., Dempsey, G. T., & Zhuang, X. (2007). Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes. Science, 317(5845), 1749â1753.

Authors 4

Mark Bates (first)
Bo Huang (additional)
Graham T. Dempsey (additional)
Xiaowei Zhuang (additional)

References 33 Referenced 1,311

L. Novotny B. Hecht Principles of Nano-optics (Cambridge Univ. Press Cambridge 2006). (10.1017/CBO9780511813535)
10.1126/science.1137395
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10.1103/PhysRevLett.94.108101
10.1021/ja044686x
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10.1073/pnas.75.4.1820
10.1083/jcb.108.2.389
10.1016/j.copbio.2004.12.003
We thank M. Rust for initial discussions of this work W. Wang for help with analysis on dye-labeled antibodies and S. Liu for providing some DNA constructs. This work was supported in part by the National Institutes of Health (grant GM 068518) and a Packard Science and Engineering Fellowship (to X.Z.). X.Z. is a Howard Hughes Medical Institute Investigator.

Dates

Type	When
Created	18 years ago (Aug. 16, 2007, 8:54 p.m.)
Deposited	1 year, 7 months ago (Jan. 10, 2024, 4:31 a.m.)
Indexed	17 minutes ago (Aug. 29, 2025, 6:22 a.m.)
Issued	17 years, 11 months ago (Sept. 21, 2007)
Published	17 years, 11 months ago (Sept. 21, 2007)
Published Print	17 years, 11 months ago (Sept. 21, 2007)

Funders 0

None

BibTeX

@article{Bates_2007, title={Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes}, volume={317}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.1146598}, DOI={10.1126/science.1146598}, number={5845}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Bates, Mark and Huang, Bo and Dempsey, Graham T. and Zhuang, Xiaowei}, year={2007}, month=sep, pages={1749–1753} }

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