Multimodal Multiphoton Imaging of the Lipid Bilayer by Dye-Based Sum-Frequency Generation and Coherent Anti-Stokes Raman Scattering
- Takaha MizuguchiTakaha MizuguchiDepartment of Pharmacology School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, JapanGraduate School of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, Kanagawa 223-8522, JapanMore by Takaha Mizuguchi,
- Atsuya MomotakeAtsuya MomotakeDepartment of Chemistry, Faculty of Pure and Applied Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8571, JapanMore by Atsuya Momotake,
- Mafumi HishidaMafumi HishidaDepartment of Chemistry, Faculty of Pure and Applied Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8571, JapanMore by Mafumi Hishida,
- Masato YasuiMasato YasuiDepartment of Pharmacology School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, JapanKeio Advanced Research Center for Water Biology and Medicine, Keio University, 2-15-45 Mita, Minato-ku, Tokyo 108-8345, JapanMore by Masato Yasui,
- Yasuhiko YamamotoYasuhiko YamamotoDepartment of Chemistry, Faculty of Pure and Applied Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8571, JapanMore by Yasuhiko Yamamoto,
- Toshiharu SaikiToshiharu SaikiGraduate School of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, Kanagawa 223-8522, JapanMore by Toshiharu Saiki, and
- Mutsuo Nuriya*Mutsuo Nuriya*Email: [email protected]Department of Pharmacology School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, JapanKeio Advanced Research Center for Water Biology and Medicine, Keio University, 2-15-45 Mita, Minato-ku, Tokyo 108-8345, JapanGraduate School of Environment and Information Sciences, Yokohama National University, Kanagawa 240-8501, JapanPrecursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), Kawaguchi, Saitama 332-0012, JapanMore by Mutsuo Nuriya
Abstract
Coherent anti-Stokes Raman scattering (CARS) imaging is widely used for imaging molecular vibrations inside cells and tissues. Lipid bilayers are potential analytes for CARS imaging due to their abundant CH2 vibrational bonds. However, identifying the plasma membrane is challenging since it possesses a thin structure and is closely apposed to lipid structures inside the cells. Since the plasma membrane provides the most prominent asymmetric location within cells, orientation sensitive sum-frequency generation (SFG) imaging is a promising technique for selective visualization of the plasma membrane labeled by a nonfluorescent and SFG-specific dye, Ap3, when using a CARS microscope system. In this study, we closely compare the characteristics of lipid bilayer imaging by dye-based SFG and CARS using giant vesicles (GVs) and N27 rat dopaminergic neural cells. As a result, we show that CARS imaging can be exploited for the visualization of whole lipid structures inside GVs and cells but is insufficient for identification of the plasma membrane, which instead can be achieved using dye-based SFG imaging. In addition, we demonstrate that these unique properties can be combined and applied to the live-cell tracking of intracellular lipid structures such as lipid droplets beneath the plasma membrane. Thus, multimodal multiphoton imaging through a combination of dye-based SFG and CARS can serve as a powerful chemical imaging tool to investigate lipid bilayers in GVs and living cells.
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This article is cited by 3 publications.
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- Takaha Mizuguchi, Mutsuo Nuriya. Applications of second harmonic generation (SHG)/sum-frequency generation (SFG) imaging for biophysical characterization of the plasma membrane. Biophysical Reviews 2020, 12 (6) , 1321-1329. https://doi.org/10.1007/s12551-020-00768-4