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Journal Article

Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes

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Hell,  Stefan W.
Optical Nanoscopy, Max Planck Institute for Medical Research, Max Planck Society;

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http://science.sciencemag.org/content/355/6325/606/tab-pdf
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https://dx.doi.org/10.1126/science.aak9913
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Citation

Balzarotti, F., Eilers, Y., Gwosch, K. C., Gynnå, A. H., Westphal, V., Stefani, F. D., et al. (2017). Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes. Science, 355(6325), 606-612. doi:10.1126/science.aak9913.


Cite as: https://hdl.handle.net/11858/00-001M-0000-002C-B784-F
Abstract
We introduce MINFLUX, a concept for localizing photon emitters in space. By probing the emitter with a local intensity minimum of excitation light, MINFLUX minimizes the fluorescence photons needed for high localization precision. A 22-fold reduction of photon detections over that required in popular centroid-localization is demonstrated. In superresolution microscopy, MINFLUX attained ~1-nm precision, resolving molecules only 6 nm apart. Tracking single fluorescent proteins by MINFLUX increased the temporal resolution and the number of localizations per trace by 100-fold, as demonstrated with diffusing 30S ribosomal subunits in living Escherichia coli As conceptual limits have not been reached, we expect this localization modality to break new ground for observing the dynamics, distribution, and structure of macromolecules in living cells and beyond.