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Real-Time Polarization-Resolved Imaging of Living Tissues Based on Two-Photon Excitation Spinning-Disk Confocal Microscopy

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Title: Real-Time Polarization-Resolved Imaging of Living Tissues Based on Two-Photon Excitation Spinning-Disk Confocal Microscopy
Authors: Goto, Ai Browse this author
Otomo, Kohei Browse this author →KAKEN DB
Nemoto, Tomomi Browse this author →KAKEN DB
Keywords: second harmonic generation
polarization-resolved optical microscopy
in vivo imaging
spinning-disk microscopy
two-photon excitation fluorescence microscopy
Issue Date: 16-Apr-2019
Publisher: Frontiers Media
Journal Title: Frontiers in physics
Volume: 7
Start Page: 56
Publisher DOI: 10.3389/fphy.2019.00056
Abstract: Laser scanning microscopy using high-peak-power ultrashort near infrared light pulses can visualize biological microstructures by utilizing non-linear optical processes, such as multi-photon excitation and sum frequency generation. Here we introduced a polarization-resolving detection methodology for a laser scanning microscopy system equipped with a spinning-disk confocal scanner. The developed system achieved high-speed intravital imaging of living tissues with resolving their signals to orthogonally polarized components. First, we applied the system to a liposomal vesicle labeled with the fluorescent lipophilic dye and con firmed the orientation map of the lipid bilayer. Next, by detecting polarization-resolved second harmonic generation signals, the structural orientations of the collagen fibers in fixed mouse tissues were visualized without exogenous or genetic fluorophore labeling. Finally, we demonstrated in vivo polarization-resolved second harmonic generation imaging of the collagen fibers in the mouse skeletal muscles at a 56 Hz temporal resolution. We expect that our developed methodology can achieve real-time visualization, thus, revealing the conformational changes of supramolecular structures in living animals.
Type: article
Appears in Collections:電子科学研究所 (Research Institute for Electronic Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 大友 康平

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