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Yet methods to quantify them simultaneously under physiologically relevant conditions, at resolutions sufficient to follow (sub)cellular processes, are still lacking. In particular, the correlation between cell state, mass, and mechanical properties during dynamic processes such as growth, differentiation, development, and disease remains poorly understood. Here, we develop experimental and theoretical approaches based on photothermally actuated microcantilevers integrated with optical microscopy to simultaneously monitor the morphology, mass, and quality (Q)\u2010factor of single mammalian cells and larger cellular systems under culture conditions, with millisecond time resolution. We find that the Q\u2010factor, a measure of mechanical energy dissipation, mirrors the mechanical properties of the cell, with the cytoplasm dominating as the largest and softest compartment. Our method reveals how cellular mechanical properties correlate with mass and depend on cell type, state, and growth. Applied to human spheroids, it further monitors how larger cellular systems regulate mechanical properties during development. Together, these measurements quantify mechanobiological parameters that have so far remained inaccessible, opening the way to a more complete characterization of how cellular systems regulate mechanics during fundamental processes of life.<\/jats:p>","DOI":"10.1002\/advs.77926","type":"journal-article","created":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T18:00:31Z","timestamp":1790532031000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Real\u2010Time Monitoring of Mass and Mechanical Properties in Single Cells and Multicellular Spheroids"],"prefix":"10.1002","author":[{"given":"Ilaria","family":"Incaviglia","sequence":"first","affiliation":[{"name":"Department of Biosystems Science and Engineering ETH Z\u00fcrich  Basel Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-5956-1052","authenticated-orcid":false,"given":"Giulia E. M.","family":"Ammirati","sequence":"additional","affiliation":[{"name":"Department of Biosystems Science and Engineering ETH Z\u00fcrich  Basel Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sophie","family":"Herzog","sequence":"additional","affiliation":[{"name":"Department of Biosystems Science and Engineering ETH Z\u00fcrich  Basel Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tommy","family":"Krug","sequence":"additional","affiliation":[{"name":"John A. Paulson School of Engineering and Applied Sciences Harvard University  Cambridge Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5985-9800","authenticated-orcid":false,"given":"Nico","family":"Strohmeyer","sequence":"additional","affiliation":[{"name":"Department of Biosystems Science and Engineering ETH Z\u00fcrich  Basel Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0867-5208","authenticated-orcid":false,"given":"Miguel","family":"Camacho Rufino","sequence":"additional","affiliation":[{"name":"Institute of Human Biology (IHB) Roche Pharma Research and Early Development (pRED), Roche Innovation Center Basel, F. Hoffmann\u2010La Roche Ltd, Basel Basel Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthias P.","family":"Lutolf","sequence":"additional","affiliation":[{"name":"Institute of Human Biology (IHB) Roche Pharma Research and Early Development (pRED), Roche Innovation Center Basel, F. Hoffmann\u2010La Roche Ltd, Basel Basel Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6678-5208","authenticated-orcid":false,"given":"David A.","family":"Weitz","sequence":"additional","affiliation":[{"name":"John A. Paulson School of Engineering and Applied Sciences Harvard University  Cambridge Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3075-0665","authenticated-orcid":false,"given":"Daniel J.","family":"M\u00fcller","sequence":"additional","affiliation":[{"name":"Department of Biosystems Science and Engineering ETH Z\u00fcrich  Basel 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