Cortex-Wide, Cellular-Resolution Volumetric Imaging with a Modular Two-Photon Imaging Platform

Cortex-Wide, Cellular-Resolution Volumetric Imaging with a Modular Two-Photon Imaging Platform | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Cortex-Wide, Cellular-Resolution Volumetric Imaging with a Modular Two-Photon Imaging Platform Bo Li, Jiahao Hu, Yanfeng Zhu, Shoupei Liu, Chengyu Li, Min Zhang, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7024540/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Mapping cortex-wide neuronal activity at single-cell resolution has been limited by the physical tradeoff 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 mm² 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 mm³, 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. Biological sciences/Neuroscience Biological sciences/Biological techniques/Imaging/Fluorescence imaging Biological sciences/Biological techniques/Microscopy/Multiphoton microscopy Biological sciences/Biological techniques/Software Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryVideo1.mp4 Meso2P System: 3D Model Visualization SupplementaryVideo2.mp4 Comparison and demonstration of parabolic lateral collection module combined with NeuroPixelAI SupplementaryVideo3.mp4 Standard processing session for large-FOV imaging data using NeuroPixelAI SupplementaryVideo4.mp4 Global dynamic transmission map at single-neuron resolution per synchronization event SupplementaryVideo5.mp4 Continuous occlusion of blood vessels caused by 5 μm MPs SupplementaryVideo6.mp4 The occlusion part of the blood vessel flows away SupplementaryVideo7.mp4 Smaller size and more NPs are widely distributed in blood vessels after injection SupplementaryVideo8.mp4 The NPs is transferred from blood vessels ro NPcell SupplementaryVideo9.mp4 NPs is repeatedly occluded to form larger particles Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7024540","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":490537389,"identity":"623bf797-a2d8-49de-9d80-5ae5b585da92","order_by":0,"name":"Bo 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