Cellular multipoint adaptive technology for two-photon mesoscope

Area of Science:

• Neuroscience
• Optical Imaging
• Biophysics

Background:

• High spatial resolution optical imaging across the entire field of view (FOV) is critical for analyzing large-scale neural circuit dynamics in neuroscience.
• Current mesoscopic imaging techniques face limitations in achieving consistent optical quality and extending the FOV beyond the objective's original design.
• Accurate spatiotemporal pattern capture of neural activity at cellular and subcellular resolution is essential for understanding brain function and disease.

Purpose of the Study:

• To develop a technology that extends the imaging FOV in two-photon mesoscopy while enhancing optical quality across the entire FOV.
• To establish a robust method for significantly extending the FOV beyond the microscope objective's original design specifications.
• To maintain original resolution specifications while improving imaging capabilities for broader neuroscience applications.

Main Methods:

• Implementation of a novel approach combining block scanning with adaptive optical (AO) correction using cellular multipoint adaptive technology (CMAT).
• Division of the imaging area into subregions, each optimized with deformable mirror (DM) compensation and real-time wavefront correction.
• User-defined sub-region scanning functions that automatically load aberration correction values for optimal optical performance.

Main Results:

• CMAT significantly enhances two-photon mesoscope imaging performance, extending the effective FOV from 6x6 mm^2 to 8x8 mm^2.
• Marked improvement in optical quality in peripheral regions, maintaining high resolution (~1 µm lateral, ~10 µm axial) centrally and improving it peripherally (~1.3 µm lateral, ~14 µm axial).
• Demonstrated enhancement in image contrast, optical resolution, and signal-to-noise ratio (SNR) for Ca2+ imaging in transgenic mouse models.

Conclusions:

• CMAT technology significantly extends the effective FOV and enhances optical quality in two-photon mesoscope systems.
• This approach provides a pivotal technical advance for large-scale functional imaging of neural circuits with cellular and subcellular resolution.
• The method offers reliable single-cell resolution imaging across extensive brain regions, advancing neuroscience research.