Fluorescence radial fluctuation enables two-photon super-resolution microscopy
Despite recent improvements in microscopy, it is still difficult to apply super-resolution microscopy for deep imaging due to the deterioration of light convergence properties in thick specimens. As a strategy to avoid such optical limitations for deep super-resolution imaging, we focused on super-r...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-10-01
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| Series: | Frontiers in Cellular Neuroscience |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fncel.2023.1243633/full |
| _version_ | 1797662936581275648 |
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| author | Motosuke Tsutsumi Motosuke Tsutsumi Taiga Takahashi Taiga Takahashi Kentaro Kobayashi Tomomi Nemoto Tomomi Nemoto Tomomi Nemoto |
| author_facet | Motosuke Tsutsumi Motosuke Tsutsumi Taiga Takahashi Taiga Takahashi Kentaro Kobayashi Tomomi Nemoto Tomomi Nemoto Tomomi Nemoto |
| author_sort | Motosuke Tsutsumi |
| collection | DOAJ |
| description | Despite recent improvements in microscopy, it is still difficult to apply super-resolution microscopy for deep imaging due to the deterioration of light convergence properties in thick specimens. As a strategy to avoid such optical limitations for deep super-resolution imaging, we focused on super-resolution radial fluctuation (SRRF), a super-resolution technique based on image analysis. In this study, we applied SRRF to two-photon microscopy (2P-SRRF) and characterized its spatial resolution, suitability for deep observation, and morphological reproducibility in real brain tissue. By the comparison with structured illumination microscopy (SIM), it was confirmed that 2P-SRRF exhibited two-point resolution and morphological reproducibility comparable to that of SIM. The improvement in spatial resolution was also demonstrated at depths of more than several hundred micrometers in a brain-mimetic environment. After optimizing SRRF processing parameters, we successfully demonstrated in vivo high-resolution imaging of the fifth layer of the cerebral cortex using 2P-SRRF. This is the first report on the application of SRRF to in vivo two-photon imaging. This method can be easily applied to existing two-photon microscopes and can expand the visualization range of super-resolution imaging studies. |
| first_indexed | 2024-03-11T19:07:19Z |
| format | Article |
| id | doaj.art-857944fc5a464006a5ac02a1fc005eaf |
| institution | Directory Open Access Journal |
| issn | 1662-5102 |
| language | English |
| last_indexed | 2024-03-11T19:07:19Z |
| publishDate | 2023-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj.art-857944fc5a464006a5ac02a1fc005eaf2023-10-10T05:27:00ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022023-10-011710.3389/fncel.2023.12436331243633Fluorescence radial fluctuation enables two-photon super-resolution microscopyMotosuke Tsutsumi0Motosuke Tsutsumi1Taiga Takahashi2Taiga Takahashi3Kentaro Kobayashi4Tomomi Nemoto5Tomomi Nemoto6Tomomi Nemoto7Biophotonics Research Group, Exploratory Research Center on Life and Living Systems, National Institutes of Natural Sciences, Okazaki, JapanResearch Division of Biophotonics, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki, JapanBiophotonics Research Group, Exploratory Research Center on Life and Living Systems, National Institutes of Natural Sciences, Okazaki, JapanResearch Division of Biophotonics, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki, JapanNikon Imaging Center, Research Institute for Electronic Science, Hokkaido University, Sapporo, JapanBiophotonics Research Group, Exploratory Research Center on Life and Living Systems, National Institutes of Natural Sciences, Okazaki, JapanResearch Division of Biophotonics, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki, JapanNikon Imaging Center, Research Institute for Electronic Science, Hokkaido University, Sapporo, JapanDespite recent improvements in microscopy, it is still difficult to apply super-resolution microscopy for deep imaging due to the deterioration of light convergence properties in thick specimens. As a strategy to avoid such optical limitations for deep super-resolution imaging, we focused on super-resolution radial fluctuation (SRRF), a super-resolution technique based on image analysis. In this study, we applied SRRF to two-photon microscopy (2P-SRRF) and characterized its spatial resolution, suitability for deep observation, and morphological reproducibility in real brain tissue. By the comparison with structured illumination microscopy (SIM), it was confirmed that 2P-SRRF exhibited two-point resolution and morphological reproducibility comparable to that of SIM. The improvement in spatial resolution was also demonstrated at depths of more than several hundred micrometers in a brain-mimetic environment. After optimizing SRRF processing parameters, we successfully demonstrated in vivo high-resolution imaging of the fifth layer of the cerebral cortex using 2P-SRRF. This is the first report on the application of SRRF to in vivo two-photon imaging. This method can be easily applied to existing two-photon microscopes and can expand the visualization range of super-resolution imaging studies.https://www.frontiersin.org/articles/10.3389/fncel.2023.1243633/fulltwo-photon microscopysuper-resolutionSRRFin vivo imagingspine morphology |
| spellingShingle | Motosuke Tsutsumi Motosuke Tsutsumi Taiga Takahashi Taiga Takahashi Kentaro Kobayashi Tomomi Nemoto Tomomi Nemoto Tomomi Nemoto Fluorescence radial fluctuation enables two-photon super-resolution microscopy Frontiers in Cellular Neuroscience two-photon microscopy super-resolution SRRF in vivo imaging spine morphology |
| title | Fluorescence radial fluctuation enables two-photon super-resolution microscopy |
| title_full | Fluorescence radial fluctuation enables two-photon super-resolution microscopy |
| title_fullStr | Fluorescence radial fluctuation enables two-photon super-resolution microscopy |
| title_full_unstemmed | Fluorescence radial fluctuation enables two-photon super-resolution microscopy |
| title_short | Fluorescence radial fluctuation enables two-photon super-resolution microscopy |
| title_sort | fluorescence radial fluctuation enables two photon super resolution microscopy |
| topic | two-photon microscopy super-resolution SRRF in vivo imaging spine morphology |
| url | https://www.frontiersin.org/articles/10.3389/fncel.2023.1243633/full |
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