Quasi-simultaneous multiplane calcium imaging of neuronal circuits

Area of Science:

• Neuroscience
• Biophysics
• Microscopy

Background:

• Two-photon excitation fluorescence microscopy is crucial for studying neuronal circuit activity.
• Standard designs limit fast imaging to a single plane, hindering 3D analysis of complex brain structures.

Purpose of the Study:

• To develop a novel quasi-simultaneous multiplane imaging technique for fast, volumetric neuronal activity recording.
• To overcome the limitations of single-plane imaging in complex 3D neuronal networks.

Main Methods:

• Developed a quasi-simultaneous multiplane imaging technique using acousto-optic deflector and static remote focusing.
• Combined hardware and software adaptable to existing two-photon microscopes.
• Enabled axial imaging at different depths without mechanical specimen or objective movement.

Main Results:

• Demonstrated high-speed, high-contrast imaging of calcium dynamics in neurons at two distinct axial planes (120 µm and 250 µm depth).
• Successfully imaged neuronal signaling in three-dimensional cortical layers of the mouse brain.

Conclusions:

• The developed technique allows for rapid, multiplane imaging of neuronal activity in 3D.
• This method enhances the study of complex neuronal networks by enabling volumetric recordings.