Ultra-large field-of-view two-photon microscopy.

Area of Science:

• Neuroscience
• Biomedical Engineering
• Optical Imaging

Background:

• High-resolution imaging of the entire murine cortex is crucial for understanding brain function.
• Existing two-photon microscopy techniques face limitations in field of view and scanning speed.
• In vivo imaging through transcranial windows presents unique challenges for optical systems.

Purpose of the Study:

• To develop and present a novel two-photon microscope capable of imaging the full extent of the murine cortex.
• To achieve objective-limited spatial resolution over a large field of view.
• To demonstrate the microscope's utility in functional brain imaging and histological analysis.

Main Methods:

• Utilized a two-photon microscopy system with a large field of view (8 mm x 10 mm).
• Employed large diameter compound lenses in the scan pathway to minimize aberrations.
• Achieved a lateral resolution of approximately 1 µm and a maximum scan speed of 5 mm/ms.
• Performed transcranial imaging of resting-state vasomotion and imaging of histological sections.

Main Results:

• The microscope successfully imaged the entire murine cortex with high spatial resolution.
• The system demonstrated a lateral resolution of ~1 µm and a scan speed of 5 mm/ms.
• Resting-state vasomotion across both cerebral hemispheres was recorded through a transcranial window.
• Histological sections were imaged efficiently without the need for image stitching.

Conclusions:

• The developed two-photon microscope offers unprecedented capabilities for large-scale, high-resolution brain imaging in mice.
• The system's performance in imaging vasomotion and histology highlights its versatility for neuroscience research.
• This technology advances the study of brain dynamics and neuropathology by overcoming previous imaging limitations.