Genetically-encoded gas vesicles bridge mesoscopic ultrasound and cellular-resolution microscopy

Genetically-encoded gas vesicles bridge mesoscopic ultrasound and cellular-resolution 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 Genetically-encoded gas vesicles bridge mesoscopic ultrasound and cellular-resolution microscopy Bettina Weigelin, Martina Giampetraglia, Lucas Freidel, Marjorie Buss, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8966722/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 Understanding complex biological systems requires imaging strategies that connect organism-level readouts with cellular resolution. Multiscale imaging approaches typically rely on combining distinct reporter systems for each modality, rather than a single genetically-encoded structure that generates multimodal contrast from the same physical source. Gas vesicles are gas-filled protein nanostructures that have been established as ultrasound contrast agents. Here we show that the same nanostructures also generate optical contrast in linear reflection microscopy and third harmonic generation microscopy. In engineered bacteria, gas vesicles produce strong third harmonic signals with cubic power dependence. In complex tissues, third harmonic signals are comparable in intensity to endogenous sources, enabling reliable detection against a physiological background. Gas vesicles remain optically detectable after phagocytosis by macrophages, enabling cellular-resolution mapping of the tissue sources underlying mesoscopic ultrasound contrast. The gas-filled architecture further confers selective sensitivity to focused multiphoton irradiation, enabling confined microscale ablation of gas vesicle positive cells and tissue regions. Reflection signals persist through aqueous tissue clearing and allow correlation of in vivo ultrasound imaging with ex vivo light-sheet microscopy of intact tumors. Together, these findings establish gas vesicles as genetically encodable reporters that connect mesoscopic ultrasound imaging with cellular-resolution microscopy and enable integrated imaging and actuation across scales. Biological sciences/Biological techniques/Imaging/Molecular imaging Biological sciences/Biological techniques/Microscopy/Light-sheet microscopy Biological sciences/Biological techniques/Microscopy/Multiphoton microscopy Biological sciences/Biological techniques/Nanobiotechnology/Nanostructures gas vesicles third harmonic generation reflection imaging light-sheet microscopy multiscale imaging Full Text Additional Declarations There is NO Competing Interest. 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. 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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-8966722","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":604604139,"identity":"987a141a-8aa1-4121-a5a1-4ab4bf9dc6e6","order_by":0,"name":"Bettina 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