Journal
NATURE BIOTECHNOLOGY
Volume 40, Issue 11, Pages 1663-+Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41587-022-01343-w
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Funding
- Hong Kong Research Grants Council [16103215, 16148816, 16102518, 16102920, T13-607/12R, T13-605/18W, C6002-17GF, N_HKUST603/19]
- Innovation and Technology Commission [ITCPD/17-9]
- Area of Excellence Scheme of the University Grants Committee [AoE/M-604/16]
- National Key R&D Program of China [2018YFE0203600]
- Guangdong Provincial Fund for Basic and Applied Basic Research [2019B1515130004]
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In this paper, we present a novel adaptive optics three-photon microscope (ALPHA-FSS) that compensates for scattering and aberrations, enabling high-resolution imaging in deep tissues. The application of this technique in the mouse brain shows promising results, with high sensitivity and precision for laser-mediated surgery.
High-resolution optical imaging deep in tissues is challenging because of optical aberrations and scattering of light caused by the complex structure of living matter. Here we present an adaptive optics three-photon microscope based on analog lock-in phase detection for focus sensing and shaping (ALPHA-FSS). ALPHA-FSS accurately measures and effectively compensates for both aberrations and scattering induced by specimens and recovers subcellular resolution at depth. A conjugate adaptive optics configuration with remote focusing enables in vivo imaging of fine neuronal structures in the mouse cortex through the intact skull up to a depth of 750 mu m below the pia, enabling near-non-invasive high-resolution microscopy in cortex. Functional calcium imaging with high sensitivity and high-precision laser-mediated microsurgery through the intact skull were also demonstrated. Moreover, we achieved in vivo high-resolution imaging of the deep cortex and subcortical hippocampus up to 1.1 mm below the pia within the intact brain. ALPHA-FSS enables high-resolution imaging deep in the mouse brain.
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