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Deep tissue multi-photon imaging using adaptive optics with direct focus sensing and shaping.

Zhongya Qin¹, Zhentao She¹, Congping Chen¹

¹Department of Electronic and Computer Engineering, The Hong Kong University of Science and Technology, Kowloon, Hong Kong, China.

Nature Biotechnology

|June 13, 2022

Summary

Researchers developed an adaptive optics microscope to overcome light scattering and aberrations for deep tissue imaging. This breakthrough enables high-resolution, near-non-invasive visualization of neuronal structures within the intact mouse brain.

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Area of Science:

Biomedical Optics
Neuroimaging
Microscopy

Background:

High-resolution optical imaging in living tissues is limited by light scattering and aberrations.
Penetrating deep into tissues for microscopy requires advanced techniques to overcome these challenges.

Purpose of the Study:

To develop a novel adaptive optics microscope for deep tissue imaging.
To enable high-resolution, in vivo visualization of neuronal structures with minimal invasiveness.

Main Methods:

An adaptive optics three-photon microscope utilizing analog lock-in phase detection for focus sensing and shaping (ALPHA-FSS) was developed.
A conjugate adaptive optics configuration with remote focusing was employed.
In vivo imaging, functional calcium imaging, and laser-mediated microsurgery were performed.

Main Results:

ALPHA-FSS effectively measured and compensated for optical aberrations and scattering.
Subcellular resolution was achieved at depths up to 750 µm in the mouse cortex through the intact skull.
High-resolution imaging extended to 1.1 mm depth in the deep cortex and hippocampus of the intact brain.

Conclusions:

The ALPHA-FSS microscope provides a near-non-invasive method for high-resolution in vivo imaging deep within the intact brain.
This technology advances neuroimaging capabilities, allowing detailed study of neuronal structures and functions at unprecedented depths.