High-speed multiplane structured illumination microscopy of living cells using an image-splitting prism
Super-resolution structured illumination microscopy (SR-SIM) can be conducted at video-rate acquisition speeds when combined with high-speed spatial light modulators and sCMOS cameras, rendering it particularly suitable for live-cell imaging. If, however, three-dimensional (3D) information is desire...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
De Gruyter
2019-12-01
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| Series: | Nanophotonics |
| Subjects: | |
| Online Access: | http://www.degruyter.com/view/j/nanoph.2020.9.issue-1/nanoph-2019-0346/nanoph-2019-0346.xml?format=INT |
| _version_ | 1818363827690930176 |
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| author | Descloux Adrien Müller Marcel Navikas Vytautas Markwirth Andreas van den Eynde Robin Lukes Tomas Hübner Wolfgang Lasser Theo Radenovic Aleksandra Dedecker Peter Huser Thomas |
| author_facet | Descloux Adrien Müller Marcel Navikas Vytautas Markwirth Andreas van den Eynde Robin Lukes Tomas Hübner Wolfgang Lasser Theo Radenovic Aleksandra Dedecker Peter Huser Thomas |
| author_sort | Descloux Adrien |
| collection | DOAJ |
| description | Super-resolution structured illumination microscopy (SR-SIM) can be conducted at video-rate acquisition speeds when combined with high-speed spatial light modulators and sCMOS cameras, rendering it particularly suitable for live-cell imaging. If, however, three-dimensional (3D) information is desired, the sequential acquisition of vertical image stacks employed by current setups significantly slows down the acquisition process. In this work, we present a multiplane approach to SR-SIM that overcomes this slowdown via the simultaneous acquisition of multiple object planes, employing a recently introduced multiplane image splitting prism combined with high-speed SIM illumination. This strategy requires only the introduction of a single optical element and the addition of a second camera to acquire a laterally highly resolved 3D image stack. We demonstrate the performance of multiplane SIM by applying this instrument to imaging the dynamics of mitochondria in living COS-7 cells. |
| first_indexed | 2024-12-13T21:54:40Z |
| format | Article |
| id | doaj.art-c0346ac8f9ae403ba9eedfd0807759a9 |
| institution | Directory Open Access Journal |
| issn | 2192-8614 |
| language | English |
| last_indexed | 2024-12-13T21:54:40Z |
| publishDate | 2019-12-01 |
| publisher | De Gruyter |
| record_format | Article |
| series | Nanophotonics |
| spelling | doaj.art-c0346ac8f9ae403ba9eedfd0807759a92022-12-21T23:30:10ZengDe GruyterNanophotonics2192-86142019-12-019114314810.1515/nanoph-2019-0346nanoph-2019-0346High-speed multiplane structured illumination microscopy of living cells using an image-splitting prismDescloux Adrien0Müller Marcel1Navikas Vytautas2Markwirth Andreas3van den Eynde Robin4Lukes Tomas5Hübner Wolfgang6Lasser Theo7Radenovic Aleksandra8Dedecker Peter9Huser Thomas10Laboratory of Nanoscale Biology, École Polytechnique Fédérale de Lausanne, 1015 Lausanne, SwitzerlandLaboratory for Nanobiology, Department of Chemistry, KU Leuven, 3000 Leuven, BelgiumLaboratory of Nanoscale Biology, École Polytechnique Fédérale de Lausanne, 1015 Lausanne, SwitzerlandBiomolecular Photonics, Department of Physics, Bielefeld University, 33501 Bielefeld, GermanyLaboratory for Nanobiology, Department of Chemistry, KU Leuven, 3000 Leuven, BelgiumLaboratory of Nanoscale Biology, École Polytechnique Fédérale de Lausanne, 1015 Lausanne, SwitzerlandBiomolecular Photonics, Department of Physics, Bielefeld University, 33501 Bielefeld, GermanyMax Planck Institute for Polymer Research, 55128 Mainz, GermanyLaboratory of Nanoscale Biology, École Polytechnique Fédérale de Lausanne, 1015 Lausanne, SwitzerlandLaboratory for Nanobiology, Department of Chemistry, KU Leuven, 3000 Leuven, BelgiumBiomolecular Photonics, Department of Physics, Bielefeld University, 33501 Bielefeld, GermanySuper-resolution structured illumination microscopy (SR-SIM) can be conducted at video-rate acquisition speeds when combined with high-speed spatial light modulators and sCMOS cameras, rendering it particularly suitable for live-cell imaging. If, however, three-dimensional (3D) information is desired, the sequential acquisition of vertical image stacks employed by current setups significantly slows down the acquisition process. In this work, we present a multiplane approach to SR-SIM that overcomes this slowdown via the simultaneous acquisition of multiple object planes, employing a recently introduced multiplane image splitting prism combined with high-speed SIM illumination. This strategy requires only the introduction of a single optical element and the addition of a second camera to acquire a laterally highly resolved 3D image stack. We demonstrate the performance of multiplane SIM by applying this instrument to imaging the dynamics of mitochondria in living COS-7 cells.http://www.degruyter.com/view/j/nanoph.2020.9.issue-1/nanoph-2019-0346/nanoph-2019-0346.xml?format=INTsuper-resolution optical microscopypacs10: 42.30.wbmultiplane image acquisitionstructured illumination microscopy |
| spellingShingle | Descloux Adrien Müller Marcel Navikas Vytautas Markwirth Andreas van den Eynde Robin Lukes Tomas Hübner Wolfgang Lasser Theo Radenovic Aleksandra Dedecker Peter Huser Thomas High-speed multiplane structured illumination microscopy of living cells using an image-splitting prism Nanophotonics super-resolution optical microscopy pacs10: 42.30.wb multiplane image acquisition structured illumination microscopy |
| title | High-speed multiplane structured illumination microscopy of living cells using an image-splitting prism |
| title_full | High-speed multiplane structured illumination microscopy of living cells using an image-splitting prism |
| title_fullStr | High-speed multiplane structured illumination microscopy of living cells using an image-splitting prism |
| title_full_unstemmed | High-speed multiplane structured illumination microscopy of living cells using an image-splitting prism |
| title_short | High-speed multiplane structured illumination microscopy of living cells using an image-splitting prism |
| title_sort | high speed multiplane structured illumination microscopy of living cells using an image splitting prism |
| topic | super-resolution optical microscopy pacs10: 42.30.wb multiplane image acquisition structured illumination microscopy |
| url | http://www.degruyter.com/view/j/nanoph.2020.9.issue-1/nanoph-2019-0346/nanoph-2019-0346.xml?format=INT |
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