Abstract
High-resolution optical imaging is critical to understanding brain function. We demonstrate that three-photon microscopy at 1,300-nm excitation enables functional imaging of GCaMP6s-labeled neurons beyond the depth limit of two-photon microscopy. We record spontaneous activity from up to 150 neurons in the hippocampal stratum pyramidale at ∼1-mm depth within an intact mouse brain. Our method creates opportunities for noninvasive recording of neuronal activity with high spatial and temporal resolution deep within scattering brain tissues.
MeSH terms
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Animals
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Brain / cytology*
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Brain / physiology
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Calmodulin / analysis
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Calmodulin / metabolism
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Green Fluorescent Proteins / analysis
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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Hippocampus / cytology
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Hippocampus / physiology
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Male
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Mice, Inbred C57BL
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Mice, Transgenic
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Microscopy, Fluorescence, Multiphoton / methods*
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Neurons / physiology*
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Recombinant Fusion Proteins / analysis
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Recombinant Fusion Proteins / metabolism
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Recombinant Proteins / analysis
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
Substances
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Calmodulin
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Recombinant Fusion Proteins
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Recombinant Proteins
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Green Fluorescent Proteins