Zhang, B. orcid.org/0009-0008-6210-9411, Daly, S. orcid.org/0000-0002-8559-3161, Zhu, C. orcid.org/0000-0002-5516-661X et al. (6 more authors) (2024) Vortex light field microscopy: 3D spectral single-molecule imaging with a twist. Optica, 11 (11). pp. 1519-1525. ISSN 2334-2536
Abstract
3D single-molecule imaging reveals nanoscale structures in cell volumes but is limited by the need for spectrally distinct fluorophores. We address this limitation with vortex light field microscopy (VLFM), a 3D spectral single-molecule localization technique with 25 nm spatial and 3 nm spectral precision over a 4 µm depth of field. By modifying our previous single-molecule light field microscope with an azimuthally oriented prism array, we generated spectral disparity orthogonal to axial disparity, enabling simultaneous spatial and spectral localization on a single detector. We demonstrate VLFM with four-color 3D single-particle tracking and two-color 3D dSTORM imaging in fixed cells, successfully identifying dyes with spectral peaks just 15 nm apart. This shows VLFM’s potential for enhancing spatial biology workflows requiring highly multiplexed imaging.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2024 The Authors. This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) > School of Mathematical and Physical Sciences |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 13 Dec 2024 10:59 |
Last Modified: | 13 Dec 2024 10:59 |
Published Version: | http://dx.doi.org/10.1364/optica.534148 |
Status: | Published |
Publisher: | Optica Publishing Group |
Refereed: | Yes |
Identification Number: | 10.1364/optica.534148 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:220354 |