Concepts for structured illumination microscopy with extended axial resolution through mirrored illumination.
| Autor: | Manton JD; Department of Chemical Engineering & Biotechnology, University of Cambridge, CB3 0AS, UK.; MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK., Ströhl F; Department of Chemical Engineering & Biotechnology, University of Cambridge, CB3 0AS, UK.; Present address: Department of Physics and Technology, UiT The Arctic University of Norway, NO-9037 Tromsø, Norway., Fiolka R; Department of Cell Biology, UT Southwestern Medical Center, 6000 Harry Hines Blvd., Dallas, Texas 75390, USA.; Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, 6000 Harry Hines Blvd., Dallas, Texas 75390, USA., Kaminski CF; Department of Chemical Engineering & Biotechnology, University of Cambridge, CB3 0AS, UK., Rees EJ; Department of Chemical Engineering & Biotechnology, University of Cambridge, CB3 0AS, UK. |
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| Jazyk: | angličtina |
| Zdroj: | Biomedical optics express [Biomed Opt Express] 2020 Mar 20; Vol. 11 (4), pp. 2098-2108. Date of Electronic Publication: 2020 Mar 20 (Print Publication: 2020). |
| DOI: | 10.1364/BOE.382398 |
| Abstrakt: | Wide-field fluorescence microscopy, while much faster than confocal microscopy, suffers from a lack of optical sectioning and poor axial resolution. 3D structured illumination microscopy (SIM) has been demonstrated to provide optical sectioning and to double the resolution limit both laterally and axially, but even with this the axial resolution is still worse than the lateral resolution of unmodified wide-field microscopy. Interferometric schemes using two high numerical aperture objectives, such as 4Pi confocal and I 5 M microscopy, have improved the axial resolution beyond that of the lateral, but at the cost of a significantly more complex optical setup. Here, we theoretically and numerically investigate a simpler dual-objective scheme which we propose can be easily added to an existing 3D-SIM microscope, providing lateral and axial resolutions in excess of 125 nm with conventional fluorophores and without the need for interferometric detection. Competing Interests: The authors declare no conflicts of interest. (Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License. Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI.) |
| Databáze: | MEDLINE |
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