Wide-field three-photon excitation in biological samples
Three-photon wide-field depth-resolved excitation is used to overcome some of the limitations in conventional point-scanning two- and three-photon microscopy. Excitation of chromophores as diverse as channelrhodopsins and quantum dots is shown, and a penetration depth of more than 700 μm into fixed...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
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Springer Nature
2018
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| Online Access: | http://hdl.handle.net/1721.1/113016 https://orcid.org/0000-0002-8261-2371 https://orcid.org/0000-0002-5738-0126 https://orcid.org/0000-0003-2220-4365 https://orcid.org/0000-0002-0419-3351 https://orcid.org/0000-0003-4698-6488 |
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| author | Fukumura, Dai Jain, Rakesh K Rowlands, Christopher Park, Demian Bruns, Oliver Thomas Piatkevich, Kiryl Bawendi, Moungi G Boyden, Edward So, Peter T. C. |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Fukumura, Dai Jain, Rakesh K Rowlands, Christopher Park, Demian Bruns, Oliver Thomas Piatkevich, Kiryl Bawendi, Moungi G Boyden, Edward So, Peter T. C. |
| author_sort | Fukumura, Dai |
| collection | MIT |
| description | Three-photon wide-field depth-resolved excitation is used to overcome some of the limitations in conventional point-scanning two- and three-photon microscopy. Excitation of chromophores as diverse as channelrhodopsins and quantum dots is shown, and a penetration depth of more than 700 μm into fixed scattering brain tissue is achieved, approximately twice as deep as that achieved using two-photon wide-field excitation. Compatibility with live animal experiments is confirmed by imaging the cerebral vasculature of an anesthetized mouse; a complete focal stack was obtained without any evidence of photodamage. As an additional validation of the utility of wide-field three-photon excitation, functional excitation is demonstrated by performing three-photon optogenetic stimulation of cultured mouse hippocampal neurons expressing a channelrhodopsin; action potentials could reliably be excited without causing photodamage. |
| first_indexed | 2024-09-23T10:00:08Z |
| format | Article |
| id | mit-1721.1/113016 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T10:00:08Z |
| publishDate | 2018 |
| publisher | Springer Nature |
| record_format | dspace |
| spelling | mit-1721.1/1130162022-09-30T18:13:53Z Wide-field three-photon excitation in biological samples Fukumura, Dai Jain, Rakesh K Rowlands, Christopher Park, Demian Bruns, Oliver Thomas Piatkevich, Kiryl Bawendi, Moungi G Boyden, Edward So, Peter T. C. Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences Massachusetts Institute of Technology. Department of Chemistry Massachusetts Institute of Technology. Department of Mechanical Engineering Massachusetts Institute of Technology. Media Laboratory McGovern Institute for Brain Research at MIT Massachusetts Institute of Technology. Center for Neurobiological Engineering Rowlands, Christopher Park, Demian Bruns, Oliver Thomas Piatkevich, Kiryl Bawendi, Moungi G Boyden, Edward So, Peter T. C. Three-photon wide-field depth-resolved excitation is used to overcome some of the limitations in conventional point-scanning two- and three-photon microscopy. Excitation of chromophores as diverse as channelrhodopsins and quantum dots is shown, and a penetration depth of more than 700 μm into fixed scattering brain tissue is achieved, approximately twice as deep as that achieved using two-photon wide-field excitation. Compatibility with live animal experiments is confirmed by imaging the cerebral vasculature of an anesthetized mouse; a complete focal stack was obtained without any evidence of photodamage. As an additional validation of the utility of wide-field three-photon excitation, functional excitation is demonstrated by performing three-photon optogenetic stimulation of cultured mouse hippocampal neurons expressing a channelrhodopsin; action potentials could reliably be excited without causing photodamage. National Institutes of Health (U.S.) (Grant NIH-5-P41-EB015871-27) National Institutes of Health (U.S.) (Grant DP3-DK101024 01) National Institutes of Health (U.S.) (Grant 1-U01-NS090438-01) National Institutes of Health (U.S.) (1-R01-HL121386-01A1) National Institutes of Health (U.S.) (Grant 1-R01-EY017656-06A1) National Institutes of Health (U.S.) (Grant 1R24MH106075) National Institutes of Health (U.S.) (Grant 2R01DA029639) National Institutes of Health (U.S.) (Grant 1R01MH103910) National Institutes of Health (U.S.) (Grant 1R01GM104948) National Science Foundation (U.S.) (Grant CBET 1053233) National Institutes of Health (U.S.) (Grant 5U54 CA151884-04) National Institutes of Health (U.S.) (Grant 9-P41-EB015871-26A1) 2018-01-08T17:03:06Z 2018-01-08T17:03:06Z 2017-05 2016-11 2018-01-05T20:37:29Z Article http://purl.org/eprint/type/JournalArticle 2047-7538 http://hdl.handle.net/1721.1/113016 Rowlands, Christopher J et al. “Wide-Field Three-Photon Excitation in Biological Samples.” Light: Science & Applications 6, 5 (November 2016): e16255 © 2017 The Author(s) https://orcid.org/0000-0002-8261-2371 https://orcid.org/0000-0002-5738-0126 https://orcid.org/0000-0003-2220-4365 https://orcid.org/0000-0002-0419-3351 https://orcid.org/0000-0003-4698-6488 http://dx.doi.org/10.1038/LSA.2016.255 Light: Science & Applications Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Springer Nature PMC |
| spellingShingle | Fukumura, Dai Jain, Rakesh K Rowlands, Christopher Park, Demian Bruns, Oliver Thomas Piatkevich, Kiryl Bawendi, Moungi G Boyden, Edward So, Peter T. C. Wide-field three-photon excitation in biological samples |
| title | Wide-field three-photon excitation in biological samples |
| title_full | Wide-field three-photon excitation in biological samples |
| title_fullStr | Wide-field three-photon excitation in biological samples |
| title_full_unstemmed | Wide-field three-photon excitation in biological samples |
| title_short | Wide-field three-photon excitation in biological samples |
| title_sort | wide field three photon excitation in biological samples |
| url | http://hdl.handle.net/1721.1/113016 https://orcid.org/0000-0002-8261-2371 https://orcid.org/0000-0002-5738-0126 https://orcid.org/0000-0003-2220-4365 https://orcid.org/0000-0002-0419-3351 https://orcid.org/0000-0003-4698-6488 |
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