Versatile high-speed volumetric imaging from microscopic to macroscopic scale by self-adaptive oblique plane microscopy

Versatile high-speed volumetric imaging from microscopic to macroscopic scale by self-adaptive oblique plane microscopy | 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 Versatile high-speed volumetric imaging from microscopic to macroscopic scale by self-adaptive oblique plane microscopy Ji Yi, Dominique Meyer, Grant Kroeschell, Xiankun Lu, Linh Hoang, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8613781/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract There is an increasing need for large-scale high-speed volumetric recording in complex multi-cellular model systems to define dynamic processes. Oblique plane microscopy (OPM) provides a solution that features oblique illumination, rapid optical scanning, and remote focusing to achieve real-time 4D microscopy. OPM implements light sheet imaging via a single primary objective lens, making the entire space below the objective accessible for large specimens, such as living mouse brain. Yet it is challenging to adopt OPM beyond a microscopic scale (i.e. size < 1 mm), limiting its broad applications. Here we present a self-adaptive OPM that leverages Abbe’s sine condition to unlock flexibility across a range of field-of-views (FOVs) (up to 8 mm 2 ) and resolutions (down to 2.2 µm 3 ). This versatility enables brain-wide single neuron volumetric calcium imaging in behaving larval zebrafish (1x0.4 mm 2 FOV at 5 Hz) and capillary blood cell tracking in living mouse brain (>3x3 mm 2 FOV) with a sweeping 0.32 mm wide volume section at 100 Hz. In optically cleared mouse brain, the flexibility allows a screen-and-zoom capability by sequentially imaging the whole brain at low-and-high magnifications to locate and resolve subcellular structures such as dendritic trees and spines. By offering a switchable imaging resolution, volume, and speed, the self-adaptive OPM achieves a versatile platform for studying a wide range of multi-cellular model system, whether in vivo or fixed and optically cleared. Biological sciences/Biological techniques/Imaging/Optical imaging Biological sciences/Neuroscience Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementalVideo4.mp4 Supplementary Video 4 SupplementalNature.docx Supplemental Materials SupplementalVideo1.mp4 Supplementary Video 1 SupplementalVideo5.mp4 Supplementary Video 5 SupplementalVideo3.mp4 Supplementary Video 3 SupplementalVideo2.mp4 Supplementary Video 2 Cite Share Download PDF Status: Under Review 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-8613781","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":590493243,"identity":"344c89d0-fe10-4e70-8eb3-bae72149d5e7","order_by":0,"name":"Ji 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Oblique plane microscopy (OPM) provides a solution that features oblique illumination, rapid optical scanning, and remote focusing to achieve real-time 4D microscopy. OPM implements light sheet imaging via a single primary objective lens, making the entire space below the objective accessible for large specimens, such as living mouse brain. Yet it is challenging to adopt OPM beyond a microscopic scale (i.e. size \u003c 1 mm), limiting its broad applications. Here we present a self-adaptive OPM that leverages Abbe’s sine condition to unlock flexibility across a range of field-of-views (FOVs) (up to 8 mm\u003csup\u003e2\u003c/sup\u003e) and resolutions (down to 2.2 µm\u003csup\u003e3\u003c/sup\u003e). 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