Photo-Induced Cell Damage Analysis for Single- and Multifocus Coherent Anti-Stokes Raman Scattering Microscopy
In this study, we investigated photo-induced damage to living cells during single- and multifocus excitations for coherent anti-Stokes Raman scattering (CARS) imaging. A near-infrared pulsed laser (709 nm) was used to induce cell damage. We compared the photo-induced cell damage in the single- and t...
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| Format: | Article |
| Language: | English |
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Wiley
2017-01-01
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| Series: | Journal of Spectroscopy |
| Online Access: | http://dx.doi.org/10.1155/2017/5725340 |
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| author | Takeo Minamikawa Yoshinori Murakami Naokazu Matsumura Hirohiko Niioka Shuichiro Fukushima Tsutomu Araki Mamoru Hashimoto |
| author_facet | Takeo Minamikawa Yoshinori Murakami Naokazu Matsumura Hirohiko Niioka Shuichiro Fukushima Tsutomu Araki Mamoru Hashimoto |
| author_sort | Takeo Minamikawa |
| collection | DOAJ |
| description | In this study, we investigated photo-induced damage to living cells during single- and multifocus excitations for coherent anti-Stokes Raman scattering (CARS) imaging. A near-infrared pulsed laser (709 nm) was used to induce cell damage. We compared the photo-induced cell damage in the single- and the multifocus excitation schemes with the condition to obtain the same CARS signal in the same frame rate. For the evaluation of cell viability, we employed 4′,6-diamidino-2-phenylindole (DAPI) fluorophores that predominantly stained the damaged cells. One- and two-photon fluorescence of DAPI fluorophores were, respectively, excited by an ultraviolet light source and the same near-infrared light source and were monitored to evaluate the cell viability during near-infrared pulsed laser irradiation. We found lower uptake of DAPI fluorophores into HeLa cells during the multifocus excitation compared with the single-focus excitation scheme in both the one- and the two-photon fluorescence examinations. This indicates a reduction of photo-induced cell damage in the multifocus excitation. Our findings suggested that the multifocus excitation scheme is expected to be suitable for CARS microscopy in terms of minimal invasiveness. |
| format | Article |
| id | doaj-art-b9a011f75cc64b9882344289d8fa0949 |
| institution | Kabale University |
| issn | 2314-4920 2314-4939 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Spectroscopy |
| spelling | doaj-art-b9a011f75cc64b9882344289d8fa09492025-02-03T01:07:30ZengWileyJournal of Spectroscopy2314-49202314-49392017-01-01201710.1155/2017/57253405725340Photo-Induced Cell Damage Analysis for Single- and Multifocus Coherent Anti-Stokes Raman Scattering MicroscopyTakeo Minamikawa0Yoshinori Murakami1Naokazu Matsumura2Hirohiko Niioka3Shuichiro Fukushima4Tsutomu Araki5Mamoru Hashimoto6Graduate School of Technology, Industrial and Social Sciences, Tokushima University, 2-1 Minami-Josanjima, Tokushima 770-8506, JapanGraduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, JapanGraduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, JapanGraduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, JapanGraduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, JapanGraduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, JapanGraduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, JapanIn this study, we investigated photo-induced damage to living cells during single- and multifocus excitations for coherent anti-Stokes Raman scattering (CARS) imaging. A near-infrared pulsed laser (709 nm) was used to induce cell damage. We compared the photo-induced cell damage in the single- and the multifocus excitation schemes with the condition to obtain the same CARS signal in the same frame rate. For the evaluation of cell viability, we employed 4′,6-diamidino-2-phenylindole (DAPI) fluorophores that predominantly stained the damaged cells. One- and two-photon fluorescence of DAPI fluorophores were, respectively, excited by an ultraviolet light source and the same near-infrared light source and were monitored to evaluate the cell viability during near-infrared pulsed laser irradiation. We found lower uptake of DAPI fluorophores into HeLa cells during the multifocus excitation compared with the single-focus excitation scheme in both the one- and the two-photon fluorescence examinations. This indicates a reduction of photo-induced cell damage in the multifocus excitation. Our findings suggested that the multifocus excitation scheme is expected to be suitable for CARS microscopy in terms of minimal invasiveness.http://dx.doi.org/10.1155/2017/5725340 |
| spellingShingle | Takeo Minamikawa Yoshinori Murakami Naokazu Matsumura Hirohiko Niioka Shuichiro Fukushima Tsutomu Araki Mamoru Hashimoto Photo-Induced Cell Damage Analysis for Single- and Multifocus Coherent Anti-Stokes Raman Scattering Microscopy Journal of Spectroscopy |
| title | Photo-Induced Cell Damage Analysis for Single- and Multifocus Coherent Anti-Stokes Raman Scattering Microscopy |
| title_full | Photo-Induced Cell Damage Analysis for Single- and Multifocus Coherent Anti-Stokes Raman Scattering Microscopy |
| title_fullStr | Photo-Induced Cell Damage Analysis for Single- and Multifocus Coherent Anti-Stokes Raman Scattering Microscopy |
| title_full_unstemmed | Photo-Induced Cell Damage Analysis for Single- and Multifocus Coherent Anti-Stokes Raman Scattering Microscopy |
| title_short | Photo-Induced Cell Damage Analysis for Single- and Multifocus Coherent Anti-Stokes Raman Scattering Microscopy |
| title_sort | photo induced cell damage analysis for single and multifocus coherent anti stokes raman scattering microscopy |
| url | http://dx.doi.org/10.1155/2017/5725340 |
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