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I will show how an in-depth description of the basic principles of diffraction-unlimited fluorescence microscopy (nanoscopy) [1-3] has spawned a new powerful superresolution concept, namely MINFLUX nanoscopy [4]. MINFLUX utilizes a local excitation intensity minimum (of a doughnut or a standing wave) that is targeted like a probe in order to localize the fluorescent molecule to be registered. In combination with single-molecule switching for sequential registration, MINFLUX [4-6] has obtained the ultimate (super)resolution: the size of a molecule. MINFLUX nanoscopy, providing 1–3 nanometer resolution in fixed and living cells, is presently being established for routine fluorescence imaging at the highest, moleculars ize resolution levels. 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.

 
[1] Hell, S.W., Wichmann, J. Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt. Lett. 19, 780-782 (1994).
[2] Hell, S.W. Far-Field Optical Nanoscopy. Science 316, 1153-1158 (2007).
[3] Hell, S.W. Microscopy and its focal switch. Nat. Methods 6, 24-32 (2009).
[4] Balzarotti, F., Eilers, Y., Gwosch, K. C., Gynnå, A. H., Westphal, V., Stefani, F. D., Elf, J., Hell, S.W. Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes. Science 355, 606-612 (2017).
[5] Eilers, Y., Ta, H., Gwosch, K. C., Balzarotti, F., Hell, S. W. MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution. PNAS 115, 6117-6122 (2018).
[6] Gwosch, K. C., Pape, J. K., Balzarotti, F., Hoess, P., Ellenberg, J., Ries, J., Hell, S. W. MINFLUX nanoscopy delivers multicolor nanometer 3D-resolution in (living) cells. (bioRxiv, doi: https://doi.org/10.1101/734251)

Stefan Hell is a director at both the Max Planck Institute for Biophysical Chemistry in Göttingen and the Max Planck Institute for Medical Research in Heidelberg, Germany.

Hell is credited with having conceived, validated and applied the first viable concept for overcoming Abbe’s diffraction-limited resolution barrier in a light-focusing fluorescence microscope. For this accomplishment he has received numerous awards, including the 2014 Kavli Prize in Nanoscience and the Nobel Prize in Chemistry.

Stefan Hell received his doctorate (1990) in physics from the University of Heidelberg. From 1991 to 1993 he worked at the European Molecular Biology Laboratory, followed by stays as a senior researcher at the University of Turku, Finland, between 1993 and 1996, and as a visiting scientist at the University of Oxford, England, in 1994. In 1997 he was appointed to the MPI for Biophysical Chemistry in Göttingen as a group leader, and was promoted to director in 2002. From 2003 to 2017 he also led a research group at the German Cancer Research Center (DKFZ). Hell holds honorary professorships in physics at the Universities of Heidelberg and Göttingen.

Photo: © Peter Badge/Typos1 in cop. with Foundation Lindau Nobel Laureate Meetings - all rights reserved 2017.

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