3D sub-diffraction imaging in a conventional confocal configuration by exploiting super-linear emitters

Denitza Denkova, Martin Ploschner, Minakshi Das, Lindsay Parker, Xianlin Zheng, Yiqing Lu, Antony Orth, Nicolle Packer, James Piper

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Sub-diffraction microscopy enables bio-imaging with unprecedented clarity. However, most super-resolution methods require complex, costly purpose-built systems, involve image post-processing and struggle with sub-diffraction imaging in 3D. Here, we realize a conceptually different super-resolution approach which circumvents these limitations and enables 3D sub-diffraction imaging on conventional confocal microscopes. We refer to it as super-linear excitation-emission (SEE) microscopy, as it relies on markers with super-linear dependence of the emission on the excitation power. Super-linear markers proposed here are upconversion nanoparticles of NaYF₄, doped with 20% Yb and unconventionally high 8% Tm, which are conveniently excited in the near-infrared biological window. We develop a computational framework calculating the 3D resolution for any viable scanning beam shape and excitation-emission probe profile. Imaging of colominic acid-coated upconversion nanoparticles endocytosed by neuronal cells, at resolutions twice better than the diffraction limit both in lateral and axial directions, illustrates the applicability of SEE microscopy for sub-cellular biology.
LanguageEnglish
Article number3695
Number of pages12
JournalNature Communications
Volume10
DOIs
Publication statusPublished - 16 Aug 2019

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Microscopy
emitters
Diffraction
Imaging techniques
Nanoparticles
Microscopic examination
configurations
microscopy
diffraction
markers
excitation
Cytology
Endocytosis
nanoparticles
clarity
Cell Biology
biology
Microscopes
microscopes
Infrared radiation

Bibliographical note

Copyright Crown 2019. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Cite this

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title = "3D sub-diffraction imaging in a conventional confocal configuration by exploiting super-linear emitters",
abstract = "Sub-diffraction microscopy enables bio-imaging with unprecedented clarity. However, most super-resolution methods require complex, costly purpose-built systems, involve image post-processing and struggle with sub-diffraction imaging in 3D. Here, we realize a conceptually different super-resolution approach which circumvents these limitations and enables 3D sub-diffraction imaging on conventional confocal microscopes. We refer to it as super-linear excitation-emission (SEE) microscopy, as it relies on markers with super-linear dependence of the emission on the excitation power. Super-linear markers proposed here are upconversion nanoparticles of NaYF₄, doped with 20{\%} Yb and unconventionally high 8{\%} Tm, which are conveniently excited in the near-infrared biological window. We develop a computational framework calculating the 3D resolution for any viable scanning beam shape and excitation-emission probe profile. Imaging of colominic acid-coated upconversion nanoparticles endocytosed by neuronal cells, at resolutions twice better than the diffraction limit both in lateral and axial directions, illustrates the applicability of SEE microscopy for sub-cellular biology.",
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3D sub-diffraction imaging in a conventional confocal configuration by exploiting super-linear emitters. / Denkova, Denitza; Ploschner, Martin; Das, Minakshi; Parker, Lindsay; Zheng, Xianlin; Lu, Yiqing; Orth, Antony; Packer, Nicolle; Piper, James.

In: Nature Communications, Vol. 10, 3695, 16.08.2019.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Zheng, Xianlin

AU - Lu, Yiqing

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AU - Packer, Nicolle

AU - Piper, James

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