Three-dimensional adaptive optical nanoscopy for thick specimen imaging at sub-50-nm resolution
- Authors
- Hao, Xiang; Allgeyer, Edward S.; Lee, Dong-Ryoung; Antonello, Jacopo; Watters, Katherine; Gerdes, Julianne A.; Schroeder, Lena K.; Bottanelli, Francesca; Zhao, Jiaxi; Kidd, Phylicia; Lessard, Mark D.; Rothman, James E.; Cooley, Lynn; Biederer, Thomas; Booth, Martin J.; Bewersdorf, Joerg
- Issue Date
- Jun-2021
- Publisher
- NATURE RESEARCH
- Citation
- NATURE METHODS, v.18, no.6, pp.688 - +
- Journal Title
- NATURE METHODS
- Volume
- 18
- Number
- 6
- Start Page
- 688
- End Page
- +
- URI
- http://scholarworks.bwise.kr/ssu/handle/2018.sw.ssu/42180
- DOI
- 10.1038/s41592-021-01149-9
- ISSN
- 1548-7091
- Abstract
- The combination of adaptive optics with an improved isoSTED nanoscope allows imaging of cells and tissues with sub-50-nm isotropic resolution. 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.
- Files in This Item
- There are no files associated with this item.
- Appears in
Collections - College of Engineering > ETC > 1. Journal Articles
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.