Scanless temporal focusing enables high-speed three-dimensional quantitative phase microscopy

Scanless temporal focusing enables high-speed three-dimensional quantitative phase 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 Scanless temporal focusing enables high-speed three-dimensional quantitative phase microscopy Peter So, Yuechuan Lin, Xiang Zhang, Rebecca Zubajlo, Zahid Yaqoob, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9270448/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 Quantitative phase microscopy (QPM) enables label-free imaging of structure and dynamics in biological and physical systems, yet achieving high-speed three-dimensional (3D) QPM with strong optical sectioning remains a central challenge. Here, we introduce a single-shot reflection-mode temporal focusing QPM (TF-QPM) that provides sub-micron optical sectioning without needs of any mechanical scanning or multiplexed acquisitions. By extending temporal focusing beyond its conventional use in multiphoton fluorescence microscopy, TF-QPM enables diffraction-limited label-free phase-sensitive volumetric imaging with 402 nm lateral and 920 nm axial resolution, markedly reduced speckle noise, and depth-resolved imaging at 3,709 Hz frame rate —an order of magnitude faster than most existing techniques and currently only limited by the camera speed. The resulting spatiotemporal phase sensitivity enables precise 3D tracking of particle motion and quantitative characterization of fast dynamics in complex and anisotropic media. For tissue imaging applications, TF-QPM achieves histology-level resolution in intact samples and supports pixel-level virtual staining, providing a rapid, label-free alternative to conventional sectioning-based workflows. Together, these results establish TF-QPM as a scanless, high-speed platform for rapid, label-free volumetric imaging across both basic research and translational applications. Physical sciences/Optics and photonics/Optical techniques/Microscopy Physical sciences/Optics and photonics/Optical techniques/Imaging and sensing Full Text Additional Declarations There is no conflict of interest Supplementary Files SupplymentaryInformation.pdf Supplementary information 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-9270448","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":618850884,"identity":"44b04a7b-546b-479b-a743-8d67083eff71","order_by":0,"name":"Peter 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Here, we introduce a single-shot reflection-mode temporal focusing QPM (TF-QPM) that provides sub-micron optical sectioning without needs of any mechanical scanning or multiplexed acquisitions. By extending temporal focusing beyond its conventional use in multiphoton fluorescence microscopy, TF-QPM enables diffraction-limited label-free phase-sensitive volumetric imaging with 402 nm lateral and 920 nm axial resolution, markedly reduced speckle noise, and depth-resolved imaging at 3,709 Hz frame rate —an order of magnitude faster than most existing techniques and currently only limited by the camera speed. The resulting spatiotemporal phase sensitivity enables precise 3D tracking of particle motion and quantitative characterization of fast dynamics in complex and anisotropic media. 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