Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Originally published in Science Express on 16 August 2007
Science 21 September 2007:
Vol. 317. no. 5845, pp. 1749 - 1753
DOI: 10.1126/science.1146598
Prev | Table of Contents | Next

Reports

Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes

Mark Bates,1 Bo Huang,2,3 Graham T. Dempsey,4 Xiaowei Zhuang2,3,5*

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.

1 School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA.
2 Howard Hughes Medical Institute, Harvard University, Cambridge, MA 02138, USA.
3 Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.
4 Graduate Program in Biophysics, Harvard University, Cambridge, MA 02138, USA.
5 Department of Physics, Harvard University, Cambridge, MA 02138, USA.

* To whom correspondence should be addressed. E-mail: zhuang{at}chemistry.harvard.edu

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Electron microscopy of whole cells in liquid with nanometer resolution.
N. d. Jonge, D. B. Peckys, G. J. Kremers, and D. W. Piston (2009)
PNAS 106, 2159-2164
   Abstract »    Full Text »    PDF »
Optical lock-in detection imaging microscopy for contrast-enhanced imaging in living cells.
G. Marriott, S. Mao, T. Sakata, J. Ran, D. K. Jackson, C. Petchprayoon, T. J. Gomez, E. Warp, O. Tulyathan, H. L. Aaron, et al. (2008)
PNAS 105, 17789-17794
   Abstract »    Full Text »    PDF »
T Cell-Dendritic Cell Immunological Synapses Contain TCR-dependent CD28-CD80 Clusters That Recruit Protein Kinase C{theta}.
S.-Y. Tseng, J. C. Waite, M. Liu, S. Vardhana, and M. L. Dustin (2008)
J. Immunol. 181, 4852-4863
   Abstract »    Full Text »    PDF »
Polarized microtubule arrays in apical dendrites and axons.
A. C. Kwan, D. A. Dombeck, and W. W. Webb (2008)
PNAS 105, 11370-11375
   Abstract »    Full Text »    PDF »
Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy.
L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. L. Gustafsson, et al. (2008)
Science 320, 1332-1336
   Abstract »    Full Text »    PDF »
Video-Rate Far-Field Optical Nanoscopy Dissects Synaptic Vesicle Movement.
V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, and S. W. Hell (2008)
Science 320, 246-249
   Abstract »    Full Text »    PDF »
Nanometer-scale mapping and single-molecule detection with color-coded nanoparticle probes.
A. Agrawal, R. Deo, G. D. Wang, M. D. Wang, and S. Nie (2008)
PNAS 105, 3298-3303
   Abstract »    Full Text »    PDF »
Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy.
B. Huang, W. Wang, M. Bates, and X. Zhuang (2008)
Science 319, 810-813
   Abstract »    Full Text »    PDF »
Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes.
H. Shroff, C. G. Galbraith, J. A. Galbraith, H. White, J. Gillette, S. Olenych, M. W. Davidson, and E. Betzig (2007)
PNAS 104, 20308-20313
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)