@article{9245ac5c370a4a8390119d50bf198e3b,
title = "Fast, cell-resolution, contiguous-wide two-photon imaging to reveal functional network architectures across multi-modal cortical areas",
abstract = "Fast and wide field-of-view imaging with single-cell resolution, high signal-to-noise ratio, and no optical aberrations have the potential to inspire new avenues of investigations in biology. However, such imaging is challenging because of the inevitable tradeoffs among these parameters. Here, we overcome these tradeoffs by combining a resonant scanning system, a large objective with low magnification and high numerical aperture, and highly sensitive large-aperture photodetectors. The result is a practically aberration-free, fast-scanning high optical invariant two-photon microscopy (FASHIO-2PM) that enables calcium imaging from a large network composed of ∼16,000 neurons at 7.5 Hz from a 9 mm2 contiguous image plane, including more than 10 sensory-motor and higher-order areas of the cerebral cortex in awake mice. Network analysis based on single-cell activities revealed that the brain exhibits small-world rather than scale-free behavior. The FASHIO-2PM is expected to enable studies on biological dynamics by simultaneously monitoring macroscopic activities and their compositional elements.",
keywords = "in vivo calcium imaging, mouse, neocortex, network analysis, objective lense, optical invariant, resonant scanning, small-world network, two-photon microscopy, wide field-of-view",
author = "Keisuke Ota and Yasuhiro Oisi and Takayuki Suzuki and Muneki Ikeda and Yoshiki Ito and Tsubasa Ito and Hiroyuki Uwamori and Kenta Kobayashi and Midori Kobayashi and Maya Odagawa and Chie Matsubara and Yoshinori Kuroiwa and Masaru Horikoshi and Junya Matsushita and Hiroyuki Hioki and Masamichi Ohkura and Junichi Nakai and Masafumi Oizumi and Atsushi Miyawaki and Toru Aonishi and Takahiro Ode and Masanori Murayama",
note = "Funding Information: We thank H. Bito, Y. Goda, H. Kamiguchi, M. Larkum, T. McHugh, S. Okabe, and C. Yokoyama for comments and discussions on the manuscript; R. Endo, R. Kato, Y. Mizukami, and K. Ueno for animal control; I. Oomoto for ImageJ program to detect blood vessels; A. Kamoshida for LabVIEW program to measure PSF; the staff of Nikon and Nikon Solutions for useful comments on the microscope; the staff of Hamamatsu Photonics K.K. for useful comments on the GaAsP PMTs; S. Itohara and K. Yamakawa for kindly providing VGAT-Cre mice; T. Nishiyama and Y. Kurokawa for comments on the videos; and the RIKEN CBS CBS-Olympus Collaboration Center and Research Resources Division for supporting our anatomical experiments and imaging acquisition. This research was supported by the AMED-Brain/Minds Project under grant numbers JP20dm0207001 (issued to A.M. and M.M.), JP19dm0207064 (issued to H.H.), and JP16dm0207041 (issued to J.N.); by JST CREST ( JPMJCR1864 and JPMJCR15E2 ) and JSPS KAKENHI ( JP18H02713 ) through grants issued to M. Oizumi; JP20H05775 (issued to M.M.); and by KAO Corp. and the Cooperative Study Program ( 237 ) of the National Institute for Physiological Sciences . Funding Information: We thank H. Bito, Y. Goda, H. Kamiguchi, M. Larkum, T. McHugh, S. Okabe, and C. Yokoyama for comments and discussions on the manuscript; R. Endo, R. Kato, Y. Mizukami, and K. Ueno for animal control; I. Oomoto for ImageJ program to detect blood vessels; A. Kamoshida for LabVIEW program to measure PSF; the staff of Nikon and Nikon Solutions for useful comments on the microscope; the staff of Hamamatsu Photonics K.K. for useful comments on the GaAsP PMTs; S. Itohara and K. Yamakawa for kindly providing VGAT-Cre mice; T. Nishiyama and Y. Kurokawa for comments on the videos; and the RIKEN CBS CBS-Olympus Collaboration Center and Research Resources Division for supporting our anatomical experiments and imaging acquisition. This research was supported by the AMED-Brain/Minds Project under grant numbers JP20dm0207001 (issued to A.M. and M.M.), JP19dm0207064 (issued to H.H.), and JP16dm0207041 (issued to J.N.); by JST CREST (JPMJCR1864 and JPMJCR15E2) and JSPS KAKENHI (JP18H02713) through grants issued to M. Oizumi; JP20H05775 (issued to M.M.); and by KAO Corp. and the Cooperative Study Program (237) of the National Institute for Physiological Sciences. M.M. designed the study. T.O. Y.K. M.M. and A.M. designed the microscope. M.H. developed the acquisition software. J.M. designed the GaAsP PMTs. K.O. performed the in vivo two-photon imaging. K.O. T.S. M.K. Y.O. and H.U. performed the viral injections and prepared the cranial windows. M. Odagawa, H.H. and K.O. performed the anatomical studies. M. Ohkura and J.N. created the G-CaMP7.09. C.M. and Y.O. performed the cloning for the AAVs. K.K. produced the AAVs. T.I. T.A. and K.O. performed cell detection using LCCD. M.I. Y.I. M. Oizumi, and K.O. performed the data analysis. M.M. T.O. A.M. K.O. T.A. Y.I. and M. Oizumi prepared the manuscript. All authors contributed to the discussion of the experimental procedures, the results, and the manuscript. T.O. is a founder of FOV Corporation. J.M. is an employee of Hamamatsu Photonics. K.K. Y.K. and M.H. are employees of Nikon Corporation. Pending patent JP, 2020-031954 (FOV Corporation and RIKEN). Publisher Copyright: {\textcopyright} 2021 Elsevier Inc.",
year = "2021",
month = jun,
day = "2",
doi = "10.1016/j.neuron.2021.03.032",
language = "English",
volume = "109",
pages = "1810--1824.e9",
journal = "Neuron",
issn = "0896-6273",
publisher = "Cell Press",
number = "11",
}