High-throughput two-photon volumetric brain imaging in freely moving mice

Area of Science:

• Neuroscience
• Biomedical Engineering
• Microscopy

Background:

• Imaging neural activity in large volumes at subcellular resolution while allowing natural animal behavior is challenging.
• Existing methods often restrict animal movement or lack the necessary scale and resolution.

Purpose of the Study:

• To develop a high-throughput miniature Bessel-beam two-photon microscope (miniBB2p) for advanced neural imaging.
• To enable imaging of neural activity across large volumes at subcellular resolution in freely moving animals.

Main Methods:

• Development of a miniature Bessel-beam two-photon microscope (miniBB2p).
• Utilized Bessel beam illumination for deep tissue penetration and reduced scattering.
• Demonstrated imaging of calcium dynamics in neurons and dendrites over a 420 × 420 × 80 μm³ volume.

Main Results:

• Successfully performed large-scale recordings of calcium activities from over 1000 neurons simultaneously.
• Imaged neural activity in the anterior cingulate cortex and secondary motor cortex of freely moving mice.
• Demonstrated miniBB2p's capability across various behavioral paradigms.

Conclusions:

• The miniBB2p facilitates precise monitoring of neural activity across large populations in a 3D volume in freely moving animals.
• This technology advances the design of miniature multiphoton microscopes for neuroscience research.
• The miniBB2p system opens new avenues for studying brain function during complex behaviors.