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MINFLUX nanoscopy delivers multicolor nanometer 3D-resolution in (living) cells

View ORCID ProfileKlaus C. Gwosch, View ORCID ProfileJasmin K. Pape, View ORCID ProfileFrancisco Balzarotti, View ORCID ProfilePhilipp Hoess, View ORCID ProfileJan Ellenberg, View ORCID ProfileJonas Ries, View ORCID ProfileStefan W. Hell
doi: https://doi.org/10.1101/734251
Klaus C. Gwosch
1Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
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  • ORCID record for Klaus C. Gwosch
Jasmin K. Pape
1Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
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Francisco Balzarotti
1Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
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Philipp Hoess
2Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
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Jan Ellenberg
2Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
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Jonas Ries
2Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
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Stefan W. Hell
1Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
3Department of Optical Nanoscopy, Max Planck Institute for Medical Research, Heidelberg, Germany
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  • For correspondence: shell@gwdg.de
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Abstract

The ultimate goal of biological superresolution fluorescence microscopy is to provide three-dimensional resolution at the size scale of a fluorescent marker. Here, we show that, by localizing individual switchable fluorophores with a probing doughnut-shaped excitation beam, MINFLUX nanoscopy provides 1–3 nanometer resolution in fixed and living cells. This progress has been facilitated by approaching each fluorophore iteratively with the probing doughnut minimum, making the resolution essentially uniform and isotropic over scalable fields of view. MINFLUX imaging of nuclear pore complexes of a mammalian cell shows that this true nanometer scale resolution is obtained in three dimensions and in two color channels. Relying on fewer detected photons than popular camera-based localization, MINFLUX nanoscopy is poised to open a new chapter in the imaging of protein complexes and distributions in fixed and living cells.

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Posted August 13, 2019.
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MINFLUX nanoscopy delivers multicolor nanometer 3D-resolution in (living) cells
Klaus C. Gwosch, Jasmin K. Pape, Francisco Balzarotti, Philipp Hoess, Jan Ellenberg, Jonas Ries, Stefan W. Hell
bioRxiv 734251; doi: https://doi.org/10.1101/734251
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MINFLUX nanoscopy delivers multicolor nanometer 3D-resolution in (living) cells
Klaus C. Gwosch, Jasmin K. Pape, Francisco Balzarotti, Philipp Hoess, Jan Ellenberg, Jonas Ries, Stefan W. Hell
bioRxiv 734251; doi: https://doi.org/10.1101/734251

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