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Removing Orientation-Induced Localization Biases in Single-Molecule Microscopy Using a Broadband Metasurface Mask

Nanoscale localization of single molecules is a crucial function in several advanced microscopy techniques, including single-molecule tracking and wide-field super-resolution imaging (1). To date, a central consideration of such techniques is how to optimize the precision of molecular localization....

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Autores principales: Backlund, Mikael P., Arbabi, Amir, Petrov, Petar N., Arbabi, Ehsan, Saurabh, Saumya, Faraon, Andrei, Moerner, W. E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5001689/
https://www.ncbi.nlm.nih.gov/pubmed/27574529
http://dx.doi.org/10.1038/nphoton.2016.93
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author Backlund, Mikael P.
Arbabi, Amir
Petrov, Petar N.
Arbabi, Ehsan
Saurabh, Saumya
Faraon, Andrei
Moerner, W. E.
author_facet Backlund, Mikael P.
Arbabi, Amir
Petrov, Petar N.
Arbabi, Ehsan
Saurabh, Saumya
Faraon, Andrei
Moerner, W. E.
author_sort Backlund, Mikael P.
collection PubMed
description Nanoscale localization of single molecules is a crucial function in several advanced microscopy techniques, including single-molecule tracking and wide-field super-resolution imaging (1). To date, a central consideration of such techniques is how to optimize the precision of molecular localization. However, as these methods continue to push toward the nanometre size scale, an increasingly important concern is the localization accuracy. In particular, single fluorescent molecules emit with an anisotropic radiation pattern of an oscillating electric dipole, which can cause significant localization biases using common estimators (2-5). Here we present the theory and experimental demonstration of a solution to this problem based on azimuthal filtering in the Fourier plane of the microscope. We do so using a high efficiency dielectric metasurface polarization/phase device composed of nanoposts with sub-wavelength spacing (6). The method is demonstrated both on fluorophores embedded in a polymer matrix, and in dL5 protein complexes that bind Malachite green (7, 8).
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spelling pubmed-50016892016-11-16 Removing Orientation-Induced Localization Biases in Single-Molecule Microscopy Using a Broadband Metasurface Mask Backlund, Mikael P. Arbabi, Amir Petrov, Petar N. Arbabi, Ehsan Saurabh, Saumya Faraon, Andrei Moerner, W. E. Nat Photonics Article Nanoscale localization of single molecules is a crucial function in several advanced microscopy techniques, including single-molecule tracking and wide-field super-resolution imaging (1). To date, a central consideration of such techniques is how to optimize the precision of molecular localization. However, as these methods continue to push toward the nanometre size scale, an increasingly important concern is the localization accuracy. In particular, single fluorescent molecules emit with an anisotropic radiation pattern of an oscillating electric dipole, which can cause significant localization biases using common estimators (2-5). Here we present the theory and experimental demonstration of a solution to this problem based on azimuthal filtering in the Fourier plane of the microscope. We do so using a high efficiency dielectric metasurface polarization/phase device composed of nanoposts with sub-wavelength spacing (6). The method is demonstrated both on fluorophores embedded in a polymer matrix, and in dL5 protein complexes that bind Malachite green (7, 8). 2016-05-16 2016 /pmc/articles/PMC5001689/ /pubmed/27574529 http://dx.doi.org/10.1038/nphoton.2016.93 Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Backlund, Mikael P.
Arbabi, Amir
Petrov, Petar N.
Arbabi, Ehsan
Saurabh, Saumya
Faraon, Andrei
Moerner, W. E.
Removing Orientation-Induced Localization Biases in Single-Molecule Microscopy Using a Broadband Metasurface Mask
title Removing Orientation-Induced Localization Biases in Single-Molecule Microscopy Using a Broadband Metasurface Mask
title_full Removing Orientation-Induced Localization Biases in Single-Molecule Microscopy Using a Broadband Metasurface Mask
title_fullStr Removing Orientation-Induced Localization Biases in Single-Molecule Microscopy Using a Broadband Metasurface Mask
title_full_unstemmed Removing Orientation-Induced Localization Biases in Single-Molecule Microscopy Using a Broadband Metasurface Mask
title_short Removing Orientation-Induced Localization Biases in Single-Molecule Microscopy Using a Broadband Metasurface Mask
title_sort removing orientation-induced localization biases in single-molecule microscopy using a broadband metasurface mask
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5001689/
https://www.ncbi.nlm.nih.gov/pubmed/27574529
http://dx.doi.org/10.1038/nphoton.2016.93
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