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

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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. In our experiments, 22 times fewer fluorescence photons are required...

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Autor principal: Balzarotti, F.
Otros Autores: Eilers, Y., Gwosch, K.C, Gynnå, A.H, Westphal, V., Stefani, F.D, Elf, J., Hell, S.W
Formato: Capítulo de libro
Lenguaje:Inglés
Publicado: American Association for the Advancement of Science 2017
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Acceso en línea:Registro en Scopus
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024 7 |2 cas  |a protein, 67254-75-5; DNA, 9007-49-2; DNA; Luminescent Proteins 
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100 1 |a Balzarotti, F. 
245 1 0 |a Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes 
260 |b American Association for the Advancement of Science  |c 2017 
270 1 0 |m Balzarotti, F.; Department of NanoBiophotonics, Max Planck Institute For Biophysical ChemistryGermany; email: francisco.balzarotti@mpibpc.mpg.de 
506 |2 openaire  |e Política editorial 
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504 |a Hell, S.W., Method of and apparatus for tracking a particle, particularly a single molecule, in a sample (2013), Patent application WO, published 23 May; Hell, S.W., High-resolution fluorescence microscopy with a structured excitation beam (2015), Patent application WO, published 2 July; Schmied, J.J., (2012) Nat. Methods, 9, pp. 1133-1134 
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520 3 |a 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. In our experiments, 22 times fewer fluorescence photons are required as compared to popular centroid localization. In superresolution microscopy, MINFLUX attained ∼1-nanometer precision, resolving molecules only 6 nanometers apart. MINFLUX tracking of single fluorescent proteins increased the temporal resolution and the number of localizations per trace by a factor of 100, 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. © 2017, American Association for the Advancement of Science. All rights reserved.  |l eng 
536 |a Detalles de la financiación: European Research Council 
536 |a Detalles de la financiación: Knut och Alice Wallenbergs Stiftelse, WO 2015/097000, WO 2013/072273 
536 |a Detalles de la financiación: We thank F. Persson for early discussions about implementations of the concept and initial tracking experiments. E. D'Este and S. J. Sahl are acknowledged for critical reading. K.C.G. and F.D.S. thank the Cusanuswerk for a stipend and the Max Planck Society for a partner group grant, respectively. A.H.G. and J.E. acknowledge the European Research Council and the Knut and Alice Wallenberg Foundation for funding. S.W.H. is inventor on patent applications WO 2013/072273 and WO 2015/097000 submitted by the Max Planck Society that cover basic principles and arrangements of MINFLUX. Further patent applications with F.B., Y.E., K.C.G., and S.W.H. as inventors have been submitted by the Max Planck Society, covering selected embodiments and procedures. S.W.H. consults and owns shares of Abberior Instruments GmbH, a manufacturer of superresolution microscopes. 
593 |a Department of NanoBiophotonics, Max Planck Institute For Biophysical Chemistry, Göttingen, Germany 
593 |a Department of Cell and Molecular Biology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden 
593 |a Centro de Investigaciones en Bionanociencias (CIBION), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina 
593 |a Departamento de Física, Facultad de Ciencias Exactas y Naturales, University of Buenos Aires, Buenos Aires, Argentina 
593 |a Department of Optical Nanoscopy, Max Planck Institute for Medical Research, Heidelberg, Germany 
593 |a Optical Nanoscopy Division, German Cancer Research Center (DKFZ), Heidelberg, Germany 
690 1 0 |a FLUORESCENT DYE 
690 1 0 |a PROTEIN 
690 1 0 |a DNA 
690 1 0 |a PHOTOPROTEIN 
690 1 0 |a CELL ORGANELLE 
690 1 0 |a CELLS AND CELL COMPONENTS 
690 1 0 |a COLIFORM BACTERIUM 
690 1 0 |a EQUIPMENT 
690 1 0 |a FLUORESCENCE 
690 1 0 |a IMAGE RESOLUTION 
690 1 0 |a LIGHT INTENSITY 
690 1 0 |a PROTEIN 
690 1 0 |a ARTICLE 
690 1 0 |a ESCHERICHIA COLI 
690 1 0 |a EXCITATION 
690 1 0 |a FLUORESCENCE 
690 1 0 |a FLUORESCENCE MICROSCOPY 
690 1 0 |a LIGHT 
690 1 0 |a MACROMOLECULE 
690 1 0 |a MOLECULAR DYNAMICS 
690 1 0 |a NANOIMAGING 
690 1 0 |a PHOTON 
690 1 0 |a PRIORITY JOURNAL 
690 1 0 |a RIBOSOME SUBUNIT 
690 1 0 |a SIMULATION 
690 1 0 |a CHEMISTRY 
690 1 0 |a FLUORESCENCE IMAGING 
690 1 0 |a NANOTECHNOLOGY 
690 1 0 |a PHOTON 
690 1 0 |a PROCEDURES 
690 1 0 |a SINGLE MOLECULE IMAGING 
690 1 0 |a SMALL RIBOSOMAL SUBUNIT 
690 1 0 |a ESCHERICHIA COLI 
690 1 0 |a DNA 
690 1 0 |a ESCHERICHIA COLI 
690 1 0 |a LUMINESCENT PROTEINS 
690 1 0 |a MICROSCOPY, FLUORESCENCE 
690 1 0 |a NANOTECHNOLOGY 
690 1 0 |a OPTICAL IMAGING 
690 1 0 |a PHOTONS 
690 1 0 |a RIBOSOME SUBUNITS, SMALL, BACTERIAL 
690 1 0 |a SINGLE MOLECULE IMAGING 
650 1 7 |2 spines  |a PRECISION 
700 1 |a Eilers, Y. 
700 1 |a Gwosch, K.C. 
700 1 |a Gynnå, A.H. 
700 1 |a Westphal, V. 
700 1 |a Stefani, F.D. 
700 1 |a Elf, J. 
700 1 |a Hell, S.W. 
773 0 |d American Association for the Advancement of Science, 2017  |g v. 355  |h pp. 606-612  |k n. 6325  |p Sci.  |x 00368075  |w (AR-BaUEN)CENRE-344  |t Science 
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856 4 0 |u https://doi.org/10.1126/science.aak9913  |y DOI 
856 4 0 |u https://hdl.handle.net/20.500.12110/paper_00368075_v355_n6325_p606_Balzarotti  |y Handle 
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999 |c 76166 

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