Deep tissue super-resolution imaging with adaptive optical two-photon multifocal structured illumination microscopy
doi: 10.1186/s43074-023-00115-2
Deep tissue super-resolution imaging with adaptive optical two-photon multifocal structured illumination microscopy
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Abstract:
Visualization of axons and dendritic spines is crucial in neuroscience research. However, traditional microscopy is limited by diffraction-limited resolution and shallow imaging depth, making it difficult to study neuronal dynamics. Two-photon multifocal structured illumination microscopy (2P-MSIM) provides super-resolution imaging along with a reasonably good penetration, but it is vulnerable to optical aberrations in deep tissues. Herein we present a novel non-inertial scanning 2P-MSIM system incorporated with adaptive optics (AO) which allows for super-resolution imaging with effective aberration correction. Our strategy is designed to correct both laser and fluorescence paths simultaneously using a spatial light modulator and a deformable mirror respectively, providing better results than the individual path corrections. The successful implementation of adaptive optical two-photon multifocal structured illumination microscopy (AO 2P-MSIM) has allowed for the super-resolution imaging of neuronal structures in a mouse brain slice at great depths and dynamic morphological characteristics of zebrafish motoneurons in vivo.
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Key words:
- Super-resolution /
- Adaptive optics /
- In vivo imaging /
- Neurons
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