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Cortex-Wide, Cellular-Resolution Volumetric Imaging with a Modular Two-Photon Imaging Platform
Summary
Researchers developed Meso2P, a modular two-photon imaging platform that achieves cortex-wide, cellular-resolution volumetric imaging by decoupling excitation and detection using a lateral paraboloid fluorescence collector. The system sustains an effective numerical aperture of 0.87 over a contiguous 6x6 mm2 field-of-view at 7.67 Hz, enabling mapping of neuronal activity across the entire cortex at single-cell resolution.
Abstract Mapping cortex-wide neuronal activity at single-cell resolution has been limited by the physical trade-off between numerical aperture and field-of-view (FOV) in two-photon microscopes. We present Meso2P, a modular two-photon platform that decouples excitation and detection by introducing a lateral paraboloid fluorescence collector. The design sustains an effective NA 0.87 over a contiguous 6 × 6 mm 2 FOV at high speed (2,048 × 2,048 pixels at 7.67 Hz). The modular platform can be upgraded with optional modules for simultaneous multi-plane imaging (1-4 planes at full resolution and speed), volumetric imaging (6 × 6 × 0.5 mm 3 , 2,048 × 2,048 × 28 voxels at 1 Hz capturing> 210,000 neurons), and holographic two-photon optogenetic stimulation for targeted perturbations. To handle the resulting large-scale data, we provide an open-source deep-learning pipeline that automates motion correction, segmentation, and spike inference. We demonstrate cortex-wide sensory responses, layer-specific network synchrony during anaesthesia, and in-vivo tracking of micro and nanoplastic distribution. Meso2P therefore provides a reproducible route to high-throughput volumetric imaging across almost the entire cortex with high detection efficiency.