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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10595032/ https://www.ncbi.nlm.nih.gov/pubmed/37881492 http://dx.doi.org/10.3389/fncel.2023.1243633 |
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author | Tsutsumi, Motosuke Takahashi, Taiga Kobayashi, Kentaro Nemoto, Tomomi |
author_facet | Tsutsumi, Motosuke Takahashi, Taiga Kobayashi, Kentaro Nemoto, Tomomi |
author_sort | Tsutsumi, Motosuke |
collection | PubMed |
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. |
format | Online Article Text |
id | pubmed-10595032 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-105950322023-10-25 Fluorescence radial fluctuation enables two-photon super-resolution microscopy Tsutsumi, Motosuke Takahashi, Taiga Kobayashi, Kentaro Nemoto, Tomomi Front Cell Neurosci Cellular Neuroscience 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. Frontiers Media S.A. 2023-10-10 /pmc/articles/PMC10595032/ /pubmed/37881492 http://dx.doi.org/10.3389/fncel.2023.1243633 Text en Copyright © 2023 Tsutsumi, Takahashi, Kobayashi and Nemoto. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Cellular Neuroscience Tsutsumi, Motosuke Takahashi, Taiga Kobayashi, Kentaro Nemoto, Tomomi Fluorescence radial fluctuation enables two-photon super-resolution microscopy |
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 | Cellular Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10595032/ https://www.ncbi.nlm.nih.gov/pubmed/37881492 http://dx.doi.org/10.3389/fncel.2023.1243633 |
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