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Updated: Oct 11, 2025
Deep-Tissue Three-Photon Fluorescence Microscopy in Intact Mouse and Zebrafish Brain
Published on: January 13, 2022
Focusing new light on brain functions: multiphoton microscopy for deep and super-resolution imaging
Hirokazu Ishii1, Kohei Otomo2, Taiga Takahashi3
1Biophotonics 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.
Two-photon microscopy offers deeper, less invasive brain imaging for studying neuron and synapse dynamics. Technological advances improve penetration depth and spatial resolution for exploring brain functions.
Area of Science:
- Neuroscience
- Biophotonics
- Microscopy
Background:
- Multiphoton microscopy, particularly two-photon microscopy, is crucial for visualizing neurobiological processes.
- Its near-infrared excitation allows deeper tissue penetration and reduced invasiveness compared to single-photon methods.
Purpose of the Study:
- To review the principles and limitations of two-photon microscopy in brain tissue imaging.
- To highlight technological advancements enhancing imaging capabilities for neuroscience research.
Main Methods:
- Review of two-photon microscopy principles.
- Analysis of technical limitations including penetration depth and spatial resolution.
- Discussion of technological improvements.
Main Results:
- Two-photon microscopy provides significant advantages for in vivo neural imaging.
- Penetration depth and spatial resolution are key limitations.
- Recent technological improvements address these limitations.
Conclusions:
- Technological advancements are expanding the utility of two-photon microscopy for deep brain imaging.
- Enhanced two-photon microscopy facilitates the investigation of previously unrevealed brain functions.
