MENU

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

Denkova, Denitza; Ploschner, Martin; Das, Minakshi; Parker, Lindsay M.; Zheng, Xianlin; Lu, Yiqing; Orth, Antony; Packer, Nicolle H.; Piper, James A.

NATURE COMMUNICATIONS
2019
VL / 10 - BP / - EP /
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 subcellular biology.

AccesS level

Gold DOAJ, Green published

MENTIONS DATA