Focusing new light on brain functions: multiphoton microscopy for deep and super-resolution imaging

Hirokazu Ishii; Kohei Otomo; Taiga Takahashi; Kazushi Yamaguchi; Tomomi Nemoto

doi:10.1016/j.neures.2021.11.011

Focusing new light on brain functions: multiphoton microscopy for deep and super-resolution imaging

Neurosci Res. 2022 Jun:179:24-30. doi: 10.1016/j.neures.2021.11.011. Epub 2021 Nov 30.

Authors

Hirokazu Ishii¹, Kohei Otomo², Taiga Takahashi³, Kazushi Yamaguchi⁴, Tomomi Nemoto⁵

Affiliations

¹ 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 (NIPS), National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi, 444-8787, Japan; Research Institute for Electronic Science, Hokkaido University, Kita 20 Nishi 10, Kita, Sapporo, Hokkaido, 001-0020, Japan.
² 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 (NIPS), National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi, 444-8787, Japan; Research Institute for Electronic Science, Hokkaido University, Kita 20 Nishi 10, Kita, Sapporo, Hokkaido, 001-0020, Japan; Graduate School of Information Science and Technology, Hokkaido University, Kita 14 Nishi 9, Kita, Sapporo, Hokkaido, 060-0814, Japan; School of Life Science, The Graduate University for Advanced Studies (SOKENDAI), Higashiyama 5-1, Myodaiji, Okazaki, Aichi, 444-8787, Japan; Graduate School of Medicine, Juntendo University, Hongo 2-1-1, Bunkyo-ku, Tokyo, 113-8421, Japan.
³ 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 (NIPS), National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi, 444-8787, Japan; Graduate School of Information Science and Technology, Hokkaido University, Kita 14 Nishi 9, Kita, Sapporo, Hokkaido, 060-0814, Japan; School of Life Science, The Graduate University for Advanced Studies (SOKENDAI), Higashiyama 5-1, Myodaiji, Okazaki, Aichi, 444-8787, Japan.
⁴ Division of Biophotonics, National Institute for Physiological Sciences (NIPS), National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi, 444-8787, Japan; Graduate School of Information Science and Technology, Hokkaido University, Kita 14 Nishi 9, Kita, Sapporo, Hokkaido, 060-0814, Japan.
⁵ 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 (NIPS), National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi, 444-8787, Japan; Research Institute for Electronic Science, Hokkaido University, Kita 20 Nishi 10, Kita, Sapporo, Hokkaido, 001-0020, Japan; Graduate School of Information Science and Technology, Hokkaido University, Kita 14 Nishi 9, Kita, Sapporo, Hokkaido, 060-0814, Japan; School of Life Science, The Graduate University for Advanced Studies (SOKENDAI), Higashiyama 5-1, Myodaiji, Okazaki, Aichi, 444-8787, Japan. Electronic address: tn@nips.ac.jp.

PMID: 34861295
DOI: 10.1016/j.neures.2021.11.011

Abstract

Multiphoton microscopy has become a powerful tool for visualizing neurobiological phenomena such as the dynamics of individual synapses and the functional activities of neurons. Owing to its near-infrared excitation laser wavelength, multiphoton microscopy achieves greater penetration depth and is less invasive than single-photon excitation. Here, we review the principles of two-photon microscopy and its technical limitations (penetration depth and spatial resolution) on brain tissue imaging. We then describe the technological improvements of two-photon microscopy that enable deeper imaging with higher spatial resolution for investigating unrevealed brain functions.

Keywords: Adaptive optics; Multiphoton excitation; Nano material; STED microscopy; Semiconductor laser; Super-resolution microscopy.

Publication types

Review

MeSH terms

Brain / diagnostic imaging
Microscopy, Fluorescence, Multiphoton* / methods
Neurons*