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Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules

Single-molecule localization microscopy enables far-field imaging with lateral resolution in the range of 10 to 20 nanometres, exploiting the fact that the centre position of a single-molecule’s image can be determined with much higher accuracy than the size of that image itself. However, attaining...

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Autores principales: Szalai, Alan M., Siarry, Bruno, Lukin, Jerónimo, Williamson, David J., Unsain, Nicolás, Cáceres, Alfredo, Pilo-Pais, Mauricio, Acuna, Guillermo, Refojo, Damián, Owen, Dylan M., Simoncelli, Sabrina, Stefani, Fernando D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7822951/
https://www.ncbi.nlm.nih.gov/pubmed/33483489
http://dx.doi.org/10.1038/s41467-020-20863-0
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author Szalai, Alan M.
Siarry, Bruno
Lukin, Jerónimo
Williamson, David J.
Unsain, Nicolás
Cáceres, Alfredo
Pilo-Pais, Mauricio
Acuna, Guillermo
Refojo, Damián
Owen, Dylan M.
Simoncelli, Sabrina
Stefani, Fernando D.
author_facet Szalai, Alan M.
Siarry, Bruno
Lukin, Jerónimo
Williamson, David J.
Unsain, Nicolás
Cáceres, Alfredo
Pilo-Pais, Mauricio
Acuna, Guillermo
Refojo, Damián
Owen, Dylan M.
Simoncelli, Sabrina
Stefani, Fernando D.
author_sort Szalai, Alan M.
collection PubMed
description Single-molecule localization microscopy enables far-field imaging with lateral resolution in the range of 10 to 20 nanometres, exploiting the fact that the centre position of a single-molecule’s image can be determined with much higher accuracy than the size of that image itself. However, attaining the same level of resolution in the axial (third) dimension remains challenging. Here, we present Supercritical Illumination Microscopy Photometric z-Localization with Enhanced Resolution (SIMPLER), a photometric method to decode the axial position of single molecules in a total internal reflection fluorescence microscope. SIMPLER requires no hardware modification whatsoever to a conventional total internal reflection fluorescence microscope and complements any 2D single-molecule localization microscopy method to deliver 3D images with nearly isotropic nanometric resolution. Performance examples include SIMPLER-direct stochastic optical reconstruction microscopy images of the nuclear pore complex with sub-20 nm axial localization precision and visualization of microtubule cross-sections through SIMPLER-DNA points accumulation for imaging in nanoscale topography with sub-10 nm axial localization precision.
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spelling pubmed-78229512021-01-29 Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules Szalai, Alan M. Siarry, Bruno Lukin, Jerónimo Williamson, David J. Unsain, Nicolás Cáceres, Alfredo Pilo-Pais, Mauricio Acuna, Guillermo Refojo, Damián Owen, Dylan M. Simoncelli, Sabrina Stefani, Fernando D. Nat Commun Article Single-molecule localization microscopy enables far-field imaging with lateral resolution in the range of 10 to 20 nanometres, exploiting the fact that the centre position of a single-molecule’s image can be determined with much higher accuracy than the size of that image itself. However, attaining the same level of resolution in the axial (third) dimension remains challenging. Here, we present Supercritical Illumination Microscopy Photometric z-Localization with Enhanced Resolution (SIMPLER), a photometric method to decode the axial position of single molecules in a total internal reflection fluorescence microscope. SIMPLER requires no hardware modification whatsoever to a conventional total internal reflection fluorescence microscope and complements any 2D single-molecule localization microscopy method to deliver 3D images with nearly isotropic nanometric resolution. Performance examples include SIMPLER-direct stochastic optical reconstruction microscopy images of the nuclear pore complex with sub-20 nm axial localization precision and visualization of microtubule cross-sections through SIMPLER-DNA points accumulation for imaging in nanoscale topography with sub-10 nm axial localization precision. Nature Publishing Group UK 2021-01-22 /pmc/articles/PMC7822951/ /pubmed/33483489 http://dx.doi.org/10.1038/s41467-020-20863-0 Text en © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Szalai, Alan M.
Siarry, Bruno
Lukin, Jerónimo
Williamson, David J.
Unsain, Nicolás
Cáceres, Alfredo
Pilo-Pais, Mauricio
Acuna, Guillermo
Refojo, Damián
Owen, Dylan M.
Simoncelli, Sabrina
Stefani, Fernando D.
Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules
title Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules
title_full Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules
title_fullStr Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules
title_full_unstemmed Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules
title_short Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules
title_sort three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7822951/
https://www.ncbi.nlm.nih.gov/pubmed/33483489
http://dx.doi.org/10.1038/s41467-020-20863-0
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