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MINSTED nanoscopy enters the Ångström localization range

MPS-Authors
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Weber,  M.
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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von der Emde,  H.
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Leutenegger,  Marcel
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Gunkel,  P.
Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Sambandan,  S.
Emeritus Group Laboratory of Neurobiology, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Khan,  T. A.
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Keller-Findeisen,  J.
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Cordes,  Volker C.
Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Hell,  S. W.       
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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s41587-022-01519-4.pdf
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Citation

Weber, M., von der Emde, H., Leutenegger, M., Gunkel, P., Sambandan, S., Khan, T. A., et al. (2022). MINSTED nanoscopy enters the Ångström localization range. Nature Biotechnology. doi:10.1038/s41587-022-01519-4.


Cite as: https://hdl.handle.net/21.11116/0000-000A-2530-6
Abstract
Super-resolution techniques have achieved localization precisions in the nanometer regime. Here we report all-optical, room temperature localization of fluorophores with precision in the Ångström range. We built on the concept of MINSTED nanoscopy where precision is increased by encircling the fluorophore with the low-intensity central region of a stimulated emission depletion (STED) donut beam while constantly increasing the absolute donut power. By blue-shifting the STED beam and separating fluorophores by on/off switching, individual fluorophores bound to a DNA strand are localized with σ = 4.7 Å, corresponding to a fraction of the fluorophore size, with only 2,000 detected photons. MINSTED fluorescence nanoscopy with single-digit nanometer resolution is exemplified by imaging nuclear pore complexes and the distribution of nuclear lamin in mammalian cells labeled by transient DNA hybridization. Because our experiments yield a localization precision σ = 2.3 Å, estimated for 10,000 detected photons, we anticipate that MINSTED will open up new areas of application in the study of macromolecular complexes in cells.