Hao, Xiang;
Allgeyer, Edward S;
Lee, Dong-Ryoung;
Antonello, Jacopo;
Watters, Katherine;
Gerdes, Julianne A;
Schroeder, Lena K;
... Bewersdorf, Joerg; + view all
(2021)
Three-dimensional adaptive optical nanoscopy for thick specimen imaging at sub-50-nm resolution.
Nature Methods
, 18
pp. 688-693.
10.1038/s41592-021-01149-9.
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Abstract
Understanding cellular organization demands the best possible spatial resolution in all three dimensions. In fluorescence microscopy, this is achieved by 4Pi nanoscopy methods that combine the concepts of using two opposing objectives for optimal diffraction-limited 3D resolution with switching fluorescent molecules between bright and dark states to break the diffraction limit. However, optical aberrations have limited these nanoscopes to thin samples and prevented their application in thick specimens. Here we have developed an improved iso-stimulated emission depletion nanoscope, which uses an advanced adaptive optics strategy to achieve sub-50-nm isotropic resolution of structures such as neuronal synapses and ring canals previously inaccessible in tissue. The adaptive optics scheme presented in this work is generally applicable to any microscope with a similar beam path geometry involving two opposing objectives to optimize resolution when imaging deep in aberrating specimens.
| Type: | Article |
|---|---|
| Title: | Three-dimensional adaptive optical nanoscopy for thick specimen imaging at sub-50-nm resolution |
| Location: | United States |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1038/s41592-021-01149-9 |
| Publisher version: | https://doi.org/10.1038/s41592-021-01149-9 |
| Language: | English |
| Additional information: | This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. |
| Keywords: | Science & Technology, Life Sciences & Biomedicine, Biochemical Research Methods, Biochemistry & Molecular Biology, STED MICROSCOPY, 4PI, OPTIMIZATION |
| UCL classification: | UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology > Clinical and Experimental Epilepsy UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/10146697 |
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