PEO-CYTOP Fluoropolymer Nanosheets as a Novel Open-Skull Window for Imaging of the Living Mouse Brain
Taiga Takahashi,
Hong Zhang,
Ryosuke Kawakami,
Kenji Yarinome,
Masakazu Agetsuma,
Junichi Nabekura,
Kohei Otomo,
Yosuke Okamura,
Tomomi Nemoto
Affiliations
Taiga Takahashi
Biophotonics Research Group, Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan; Division of Biophotonics, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan; School of Life Science, The Graduate University for Advanced Studies (SOKENDAI), Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan; Research Institute for Electronic Science, Hokkaido University, Hokkaido, Kita 20 Nishi 10, Kita-ku, Sapporo, Hokkaido 001-0020, Japan; Graduate School of Information Science and Technology Hokkaido University, Hokkaido, Kita 20 Nishi 10, Kita-ku, Sapporo, Hokkaido 001-0020, Japan
Hong Zhang
Department of Applied Chemistry, School of Engineering, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan; Micro/Nano Technology Center, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan
Ryosuke Kawakami
Research Institute for Electronic Science, Hokkaido University, Hokkaido, Kita 20 Nishi 10, Kita-ku, Sapporo, Hokkaido 001-0020, Japan; Graduate School of Information Science and Technology Hokkaido University, Hokkaido, Kita 20 Nishi 10, Kita-ku, Sapporo, Hokkaido 001-0020, Japan; Department of Molecular Medicine for Pathogenesis, Graduate School of Medicine Ehime University, Shitsukawa 454, Toon, Ehime 791-0295, Japan
Kenji Yarinome
Course of Applied Science, Graduate School of Engineering, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan
Masakazu Agetsuma
Division of Homeostatic Development, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki, 444-8585, Japan
Junichi Nabekura
School of Life Science, The Graduate University for Advanced Studies (SOKENDAI), Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan; Division of Homeostatic Development, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki, 444-8585, Japan
Kohei Otomo
Biophotonics Research Group, Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan; Division of Biophotonics, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan; School of Life Science, The Graduate University for Advanced Studies (SOKENDAI), Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan; Research Institute for Electronic Science, Hokkaido University, Hokkaido, Kita 20 Nishi 10, Kita-ku, Sapporo, Hokkaido 001-0020, Japan; Graduate School of Information Science and Technology Hokkaido University, Hokkaido, Kita 20 Nishi 10, Kita-ku, Sapporo, Hokkaido 001-0020, Japan
Yosuke Okamura
Department of Applied Chemistry, School of Engineering, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan; Micro/Nano Technology Center, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan; Course of Applied Science, Graduate School of Engineering, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan
Tomomi Nemoto
Biophotonics Research Group, Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan; Division of Biophotonics, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan; School of Life Science, The Graduate University for Advanced Studies (SOKENDAI), Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan; Research Institute for Electronic Science, Hokkaido University, Hokkaido, Kita 20 Nishi 10, Kita-ku, Sapporo, Hokkaido 001-0020, Japan; Graduate School of Information Science and Technology Hokkaido University, Hokkaido, Kita 20 Nishi 10, Kita-ku, Sapporo, Hokkaido 001-0020, Japan; Corresponding author
Summary: In vivo two-photon deep imaging with a broad field of view has revealed functional connectivity among brain regions. Here, we developed a novel observation method that utilizes a polyethylene-oxide-coated CYTOP (PEO-CYTOP) nanosheet with a thickness of ∼130 nm that exhibited a water retention effect and a hydrophilized adhesive surface. PEO-CYTOP nanosheets firmly adhered to brain surfaces, which suppressed bleeding from superficial veins. By taking advantage of the excellent optical properties of PEO-CYTOP nanosheets, we performed in vivo deep imaging in mouse brains at high resolution. Moreover, PEO-CYTOP nanosheets enabled to prepare large cranial windows, achieving in vivo imaging of neural structure and Ca2+ elevation in a large field of view. Furthermore, the PEO-CYTOP nanosheets functioned as a sealing material, even after the removal of the dura. These results indicate that this method would be suitable for the investigation of neural functions that are composed of interactions among multiple regions.
