期刊
NATURE METHODS
卷 18, 期 10, 页码 1253-+出版社
NATURE PORTFOLIO
DOI: 10.1038/s41592-021-01257-6
关键词
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资金
- EMBL Interdisciplinary Postdoc (EIPOD) Programme under Marie Sklodowska Curie Cofund Actions MSCA-COFUND-FP [664726]
- EC Marie Curie COFUND EIPOD3 interdisciplinary postdoctoral fellowships
- NARSAD Young Investigator Grant from the Brain & Behavior Research Foundation
- Chica and Heinz Schaller Research Foundation [A09N/SFB1158]
- Deutsche Forschungsgemeinschaft [P8/FOR2289, 425902099]
- European Commission [951991]
- Chan Zuckerberg Initiative [2020-225346]
- EMBL
The article introduces a minimally invasive intravital imaging technique based on three-photon excitation, adaptive optics, and electrocardiogram gating for high-resolution imaging of neurons and astrocytes in the mouse brain up to a depth of 1.4 mm.
Multiphoton microscopy has become a powerful tool with which to visualize the morphology and function of neural cells and circuits in the intact mammalian brain. However, tissue scattering, optical aberrations and motion artifacts degrade the imaging performance at depth. Here we describe a minimally invasive intravital imaging methodology based on three-photon excitation, indirect adaptive optics (AO) and active electrocardiogram gating to advance deep-tissue imaging. Our modal-based, sensorless AO approach is robust to low signal-to-noise ratios as commonly encountered in deep scattering tissues such as the mouse brain, and permits AO correction over large axial fields of view. We demonstrate near-diffraction-limited imaging of deep cortical spines and (sub)cortical dendrites up to a depth of 1.4 mm (the edge of the mouse CA1 hippocampus). In addition, we show applications to deep-layer calcium imaging of astrocytes, including fibrous astrocytes that reside in the highly scattering corpus callosum. Three-photon microscopy in combination with adaptive optics-based aberration correction and ECG-triggered gating allows high-resolution imaging of neurons and astrocytes up to a depth of 1.4 mm in the mouse brain.
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