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

*Corresponding author for this work

Research output: Contribution to journalArticle

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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.
Original languageEnglish
Article number3695
Pages (from-to)1-12
Number of pages12
JournalNature Communications
Volume10
DOIs
Publication statusPublished - 16 Aug 2019

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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.

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