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  Direct supercritical angle localization microscopy for nanometer 3D superresolution

Dasgupta, A., Deschamps, J., Matti, U., Huebner, U., Becker, J., Strauss, S., et al. (2021). Direct supercritical angle localization microscopy for nanometer 3D superresolution. Nature Communications, 12(1): 1180. doi:10.1038/s41467-021-21333-x.

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 Creators:
Dasgupta, Anindita1, Author
Deschamps, Joran1, Author
Matti, Ulf1, Author
Huebner, Uwe1, Author
Becker, Jan1, Author
Strauss, Sebastian2, Author              
Jungmann, Ralf2, Author              
Heintzmann, Rainer1, Author
Ries, Jonas1, Author
Affiliations:
1external, ou_persistent22              
2Jungmann, Ralf / Molecular Imaging and Bionanotechnology, Max Planck Institute of Biochemistry, Max Planck Society, ou_2149679              

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Free keywords: Science & Technology - Other Topics;
 Abstract: 3D single molecule localization microscopy (SMLM) is an emerging superresolution method for structural cell biology, as it allows probing precise positions of proteins in cellular structures. In supercritical angle localization microscopy (SALM), z-positions of single fluorophores are extracted from the intensity of supercritical angle fluorescence, which strongly depends on their distance to the coverslip. Here, we realize the full potential of SALM and improve its z-resolution by more than four-fold compared to the state-of-the-art by directly splitting supercritical and undercritical emission, using an ultra-high NA objective, and applying fitting routines to extract precise intensities of single emitters. We demonstrate nanometer isotropic localization precision on DNA origami structures, and on clathrin coated vesicles and microtubules in cells, illustrating the potential of SALM for cell biology. Supercritical angle localisation microscopy (SALM) allows the z-positions of single fluorophores to be extracted from the intensity of supercritical angle fluorescence. Here the authors improve the z-resolution of SALM, and report nanometre isotropic localisation precision on DNA origami structures.

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Language(s): eng - English
 Dates: 2021
 Publication Status: Published online
 Pages: 9
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Degree: -

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Title: Nature Communications
  Abbreviation : Nat. Commun.
Source Genre: Journal
 Creator(s):
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Publ. Info: London : Nature Publishing Group
Pages: - Volume / Issue: 12 (1) Sequence Number: 1180 Start / End Page: - Identifier: ISSN: 2041-1723
CoNE: https://pure.mpg.de/cone/journals/resource/2041-1723