Sealing of Immersion Deuterium Dioxide and Its Application to Signal Maintenance for Ex-Vivo and In-Vivo Multiphoton Microscopy Excited at the 1700-nm Window
Excitation at the 1700-nm window is an effective means for extending imaging depth and imaging modalities in multiphoton microscopy (MPM). To enhance multiphoton signal levels and enable deep-tissue penetration, water immersion has to be replaced by deuterium dioxide (D<sub>2</sub>O) imm...
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IEEE
2017-01-01
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| Series: | IEEE Photonics Journal |
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| Online Access: | https://ieeexplore.ieee.org/document/8013790/ |
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| author | Hongji Liu Yu Du Xiao Peng Xuechang Zhou Ping Qiu Ke Wang |
| author_facet | Hongji Liu Yu Du Xiao Peng Xuechang Zhou Ping Qiu Ke Wang |
| author_sort | Hongji Liu |
| collection | DOAJ |
| description | Excitation at the 1700-nm window is an effective means for extending imaging depth and imaging modalities in multiphoton microscopy (MPM). To enhance multiphoton signal levels and enable deep-tissue penetration, water immersion has to be replaced by deuterium dioxide (D<sub>2</sub>O) immersion to boost transmittance at the 1700-nm window. The key problem facing this D<sub>2</sub>O immersion technique is the hygroscopic nature of D<sub>2</sub>O, which leads to decrease of MPM signals as time lapses. Here, we demonstrate a simple, yet very effective technique to isolate D<sub>2 </sub>O from the ambient environment, by sealing it with the paraffin liquid. We demonstrate the application of this technique to MPM signal maintenance in both three-photon fluorescence generation in a fluorescent dye and third-harmonic generation (THG) imaging of biological tissue, excited at the 1700-nm window. <italic>Ex-vivo</italic> imaging results show that during an imaging session of 5 h, multiphoton signals of both modalities can be maintained with no deterioration due to absorption of water vapor from the environment. Furthermore, we demonstrate <italic> in-vivo</italic> deep-tissue mouse brain imaging using this technique, in which THG signals can be maintained for at least 5 h. This justifies the applicability and effectiveness of our D<sub>2</sub>O sealing technique for long-span <italic>in-vivo</italic> imaging. |
| format | Article |
| id | doaj-art-3302a07925404ac6b705630ebb2a362a |
| institution | DOAJ |
| issn | 1943-0655 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | IEEE |
| record_format | Article |
| series | IEEE Photonics Journal |
| spelling | doaj-art-3302a07925404ac6b705630ebb2a362a2025-08-20T03:14:51ZengIEEEIEEE Photonics Journal1943-06552017-01-01951810.1109/JPHOT.2017.27370128013790Sealing of Immersion Deuterium Dioxide and Its Application to Signal Maintenance for Ex-Vivo and In-Vivo Multiphoton Microscopy Excited at the 1700-nm WindowHongji Liu0Yu Du1Xiao Peng2Xuechang Zhou3Ping Qiu4Ke Wang5Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic Engineering, Shenzhen University, Shenzhen, ChinaKey Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic Engineering, Shenzhen University, Shenzhen, ChinaKey Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic Engineering, Shenzhen University, Shenzhen, ChinaCollege of Chemistry and Environmental Engineering, Shenzhen University, Shenzhen, ChinaCollege of Physics and Energy, Shenzhen University, Shenzhen, ChinaKey Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic Engineering, Shenzhen University, Shenzhen, ChinaExcitation at the 1700-nm window is an effective means for extending imaging depth and imaging modalities in multiphoton microscopy (MPM). To enhance multiphoton signal levels and enable deep-tissue penetration, water immersion has to be replaced by deuterium dioxide (D<sub>2</sub>O) immersion to boost transmittance at the 1700-nm window. The key problem facing this D<sub>2</sub>O immersion technique is the hygroscopic nature of D<sub>2</sub>O, which leads to decrease of MPM signals as time lapses. Here, we demonstrate a simple, yet very effective technique to isolate D<sub>2 </sub>O from the ambient environment, by sealing it with the paraffin liquid. We demonstrate the application of this technique to MPM signal maintenance in both three-photon fluorescence generation in a fluorescent dye and third-harmonic generation (THG) imaging of biological tissue, excited at the 1700-nm window. <italic>Ex-vivo</italic> imaging results show that during an imaging session of 5 h, multiphoton signals of both modalities can be maintained with no deterioration due to absorption of water vapor from the environment. Furthermore, we demonstrate <italic> in-vivo</italic> deep-tissue mouse brain imaging using this technique, in which THG signals can be maintained for at least 5 h. This justifies the applicability and effectiveness of our D<sub>2</sub>O sealing technique for long-span <italic>in-vivo</italic> imaging.https://ieeexplore.ieee.org/document/8013790/Fluorescence microscopynon-linear microscopyoptical properties of photonic materials. |
| spellingShingle | Hongji Liu Yu Du Xiao Peng Xuechang Zhou Ping Qiu Ke Wang Sealing of Immersion Deuterium Dioxide and Its Application to Signal Maintenance for Ex-Vivo and In-Vivo Multiphoton Microscopy Excited at the 1700-nm Window IEEE Photonics Journal Fluorescence microscopy non-linear microscopy optical properties of photonic materials. |
| title | Sealing of Immersion Deuterium Dioxide and Its Application to Signal Maintenance for Ex-Vivo and In-Vivo Multiphoton Microscopy Excited at the 1700-nm Window |
| title_full | Sealing of Immersion Deuterium Dioxide and Its Application to Signal Maintenance for Ex-Vivo and In-Vivo Multiphoton Microscopy Excited at the 1700-nm Window |
| title_fullStr | Sealing of Immersion Deuterium Dioxide and Its Application to Signal Maintenance for Ex-Vivo and In-Vivo Multiphoton Microscopy Excited at the 1700-nm Window |
| title_full_unstemmed | Sealing of Immersion Deuterium Dioxide and Its Application to Signal Maintenance for Ex-Vivo and In-Vivo Multiphoton Microscopy Excited at the 1700-nm Window |
| title_short | Sealing of Immersion Deuterium Dioxide and Its Application to Signal Maintenance for Ex-Vivo and In-Vivo Multiphoton Microscopy Excited at the 1700-nm Window |
| title_sort | sealing of immersion deuterium dioxide and its application to signal maintenance for ex vivo and in vivo multiphoton microscopy excited at the 1700 nm window |
| topic | Fluorescence microscopy non-linear microscopy optical properties of photonic materials. |
| url | https://ieeexplore.ieee.org/document/8013790/ |
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