{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T13:00:32Z","timestamp":1790600432462,"version":"4.1.0"},"reference-count":35,"publisher":"American Chemical Society (ACS)","issue":"9","license":[{"start":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T00:00:00Z","timestamp":1775779200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100014168","name":"University of Melbourne","doi-asserted-by":"publisher","award":["GPUM\/24\/010"],"award-info":[{"award-number":["GPUM\/24\/010"]}],"id":[{"id":"10.13039\/501100014168","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004040","name":"KU Leuven","doi-asserted-by":"publisher","award":["C14\/22\/085"],"award-info":[{"award-number":["C14\/22\/085"]}],"id":[{"id":"10.13039\/501100004040","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004040","name":"KU Leuven","doi-asserted-by":"publisher","award":["GPUM\/24\/010"],"award-info":[{"award-number":["GPUM\/24\/010"]}],"id":[{"id":"10.13039\/501100004040","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004040","name":"KU Leuven","doi-asserted-by":"publisher","award":["IDN\/20\/021"],"award-info":[{"award-number":["IDN\/20\/021"]}],"id":[{"id":"10.13039\/501100004040","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003130","name":"Fonds Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","award":["11A0S25N"],"award-info":[{"award-number":["11A0S25N"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003130","name":"Fonds Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","award":["12AGZ24N"],"award-info":[{"award-number":["12AGZ24N"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003130","name":"Fonds Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","award":["12AML24N"],"award-info":[{"award-number":["12AML24N"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003130","name":"Fonds Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","award":["1S95125N"],"award-info":[{"award-number":["1S95125N"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003130","name":"Fonds Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","award":["G0C2422N"],"award-info":[{"award-number":["G0C2422N"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The viscosity of the cytoplasm plays a key role in regulating molecular diffusion and cellular mechanics, yet quantifying it in living cells remains technically challenging. Genetically encoded multimeric nanoparticles (GEMs) have emerged as useful probes for intracellular microrheology; however, current analyses rely on single-particle tracking, which is limited by probe density, imaging noise, and expression variability. Here, we combine GEMs with differential dynamic microscopy (DDM) to enable quantitative, noninvasive, and rapid measurement of intracellular viscosity using standard wide-field fluorescence imaging. DDM extracts particle dynamics from ensemble spatiotemporal intensity fluctuations, yielding diffusion coefficients and viscosity estimates, even in crowded or heterogeneous environments where tracking fails. Validation with fluorescent nanoparticles diffusing in water confirmed that DDM accurately reproduced theoretical viscosities across a wide range of particle sizes and concentrations. Comparison with single-particle tracking (SPT) demonstrated equivalent precision under dilute conditions and improved the performance under crowding. To showcase the potential of this approach, we applied GEM-DDM to colorectal cancer cell lines with different metastatic potentials. Cytoplasmic viscosity correlated with aggressiveness, increasing from 1.9 to 2.3 cP in poorly metastatic lines to 3.6\u20133.7 cP in highly metastatic lines, consistent with greater macromolecular crowding and cytoplasmic reorganization reported in aggressive cells. Together, these results establish GEM-DDM as a fast, reproducible, and accessible platform for intracellular microrheology to link the physical state of the cytoplasm to cell function and disease progression.<\/jats:p>","DOI":"10.1021\/cbmi.5c00261","type":"journal-article","created":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T05:54:04Z","timestamp":1775800444000},"page":"1978-1986","source":"Crossref","is-referenced-by-count":0,"title":["Quantitative Analysis\nof Cytoplasmic Viscosity in\nColorectal Cancer Cells by Differential Dynamic Microscopy of Genetically\nEncoded Nanoparticles"],"prefix":"10.1021","volume":"4","author":[{"given":"Steven","family":"Huysecom","sequence":"first","affiliation":[{"name":"KU Leuven , , ,","place":["Leuven, Belgium, B-3001"]},{"name":"Ghent University , , ,","place":["Zwijnaarde, Belgium, B-9052"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-9577-5226","authenticated-orcid":true,"given":"Kiki","family":"Schouwstra","sequence":"additional","affiliation":[{"name":"KU Leuven , , ,","place":["Leuven, Belgium, B-3001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samet","family":"Aytekin","sequence":"additional","affiliation":[{"name":"KU Leuven , , ,","place":["Leuven, Belgium, B-3001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalie","family":"Neyrinck","sequence":"additional","affiliation":[{"name":"KU Leuven , , ,","place":["Leuven, Belgium, B-3001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-9648-8052","authenticated-orcid":true,"given":"Yana","family":"Heyvaert","sequence":"additional","affiliation":[{"name":"KU Leuven , , ,","place":["Leuven, Belgium, B-3001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2882-6907","authenticated-orcid":true,"given":"Boris","family":"Louis","sequence":"additional","affiliation":[{"name":"KU Leuven , , ,","place":["Leuven, Belgium, B-3001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1258-9396","authenticated-orcid":true,"given":"Susana","family":"Rocha","sequence":"additional","affiliation":[{"name":"KU Leuven , , ,","place":["Leuven, Belgium, B-3001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4785-0040","authenticated-orcid":true,"given":"Guillermo","family":"Soli\u0301s-Fernandez","sequence":"additional","affiliation":[{"name":"KU Leuven , , ,","place":["Leuven, Belgium, B-3001"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,4,10]]},"reference":[{"key":"2026092808072424600_cit1","doi-asserted-by":"publisher","first-page":"89794","DOI":"10.7554\/eLife.89794","article-title":"Diffusive\nLensing as a Mechanism of Intracellular Transport and Compartmentalization","volume":"12","author":"Raja Venkatesh","year":"2024","journal-title":"eLife"},{"issue":"1","key":"2026092808072424600_cit2","doi-asserted-by":"publisher","first-page":"4104","DOI":"10.1038\/s41598-019-40550-5","article-title":"Cargo Diffusion Shortens\nSingle-Kinesin Runs at Low Viscous Drag","volume":"9","author":"Wilson","year":"2019","journal-title":"Sci.\nRep."},{"issue":"6","key":"2026092808072424600_cit3","doi-asserted-by":"publisher","first-page":"061013","DOI":"10.1115\/1.4039897","article-title":"Internal\nViscosity-Dependent Margination of Red Blood Cells in Microfluidic\nChannels","volume":"140","author":"Ahmed","year":"2018","journal-title":"J. Biomech. Eng."},{"issue":"3","key":"2026092808072424600_cit4","doi-asserted-by":"publisher","first-page":"563","DOI":"10.1172\/JCI109888","article-title":"Analysis of Factors\nRegulating Erythrocyte Deformability","volume":"66","author":"Mohandas","year":"1980","journal-title":"J. Clin.\nInvest."},{"issue":"1","key":"2026092808072424600_cit5","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41598-019-43200-y","article-title":"Impact of Surface-Area-to-Volume\nRatio, Internal Viscosity and Membrane Viscoelasticity on Red Blood\nCell Deformability Measured in Isotonic Condition","volume":"9","author":"Renoux","year":"2019","journal-title":"Scientific Reports 2019 9:1"},{"issue":"11","key":"2026092808072424600_cit6","doi-asserted-by":"publisher","first-page":"4130","DOI":"10.1016\/j.actbio.2012.07.011","article-title":"Optical Measurement of Biomechanical\nProperties of\nIndividual Erythrocytes from a Sickle Cell Patient","volume":"8","author":"Byun","year":"2012","journal-title":"Acta Biomater."},{"key":"2026092808072424600_cit7","doi-asserted-by":"publisher","first-page":"328","DOI":"10.1091\/mbc.e04-06-0485","article-title":"Intracellular\nMechanics of Migrating Fibroblasts \u25a1 D","volume":"16","author":"Kole","year":"2005","journal-title":"Mol. Biol. Cell"},{"issue":"29","key":"2026092808072424600_cit8","doi-asserted-by":"publisher","first-page":"4663","DOI":"10.1039\/D1CC07235H","article-title":"Assessment of Cancer Cell Migration Using a Viscosity-Sensitive\nFluorescent Probe","volume":"58","author":"Pan","year":"2022","journal-title":"Chem. Commun."},{"issue":"4","key":"2026092808072424600_cit9","doi-asserted-by":"publisher","first-page":"740","DOI":"10.1111\/febs.17306","article-title":"Viscosity\nRegulates Cell Spreading and Cell-Extracellular Matrix Interactions","volume":"292","author":"Xiao","year":"2025","journal-title":"FEBS Journal"},{"issue":"2","key":"2026092808072424600_cit10","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1006\/abbi.1997.9910","article-title":"Cell Type\nand Spatial Location Dependence of Cytoplasmic\nViscosity Measured by Time-Resolved Fluorescence Microscopy","volume":"340","author":"Srivastava","year":"1997","journal-title":"Arch. Biochem. Biophys."},{"issue":"1","key":"2026092808072424600_cit11","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1038\/nchem.120","article-title":"Imaging Intracellular\nViscosity of a Single Cell during Photoinduced Cell Death","volume":"1","author":"Kuimova","year":"2009","journal-title":"Nature Chemistry 2009 1:1"},{"issue":"1","key":"2026092808072424600_cit12","doi-asserted-by":"publisher","first-page":"316","DOI":"10.1529\/biophysj.106.099267","article-title":"Probing the\nNanoscale Viscoelasticity\nof Intracellular Fluids in Living Cells","volume":"93","author":"Guigas","year":"2007","journal-title":"Biophys.\nJ."},{"issue":"C","key":"2026092808072424600_cit13","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1016\/0076-6879(86)27020-2","article-title":"[17] Determining\nIntracellular Viscosity from the Rotational Motion of Spin Labels","volume":"127","author":"Morse","year":"1986","journal-title":"Methods Enzymol."},{"key":"2026092808072424600_cit14","doi-asserted-by":"publisher","first-page":"10134","DOI":"10.1038\/ncomms10134","article-title":"Local Viscoelasticity\nof Living Cells Measured by Rotation Magnetic Spectroscopymagnetic\nSpectroscopy","volume":"7","author":"Berret","year":"2016","journal-title":"Nat. Commun."},{"key":"2026092808072424600_cit15","doi-asserted-by":"publisher","first-page":"245","DOI":"10.1016\/B978-0-12-388422-0.00010-8","article-title":"Image Correlation\nSpectroscopy: Mapping Correlations in Space, Time, and Reciprocal\nSpace","volume":"518","author":"Wiseman","year":"2013","journal-title":"Methods Enzymol."},{"issue":"10","key":"2026092808072424600_cit16","doi-asserted-by":"publisher","first-page":"969","DOI":"10.1038\/s41592-019-0543-3","article-title":"Brillouin\nMicroscopy: An Emerging Tool for Mechanobiology","volume":"16","author":"Prevedel","year":"2019","journal-title":"Nat. Methods"},{"key":"2026092808072424600_cit17","doi-asserted-by":"publisher","DOI":"10.1101\/2023.05.28.542585","article-title":"Imaging the Microscopic Viscoelastic\nAnisotropy in Living Cells","author":"Keshmiri","year":"2023","journal-title":"bioRxiv"},{"issue":"2","key":"2026092808072424600_cit18","doi-asserted-by":"publisher","first-page":"338","DOI":"10.1016\/j.cell.2018.05.042","article-title":"MTORC1\nControls Phase Separation and the Biophysical Properties of the Cytoplasm\nby Tuning Crowding","volume":"174","author":"Delarue","year":"2018","journal-title":"Cell"},{"key":"2026092808072424600_cit19","doi-asserted-by":"publisher","DOI":"10.1101\/2024.07.05.602291","article-title":"Development\nand Characterization of 50 Nanometer Diameter Genetically Encoded\nMultimeric Nanoparticles","author":"Hernandez","year":"2024","journal-title":"bioRxiv"},{"key":"2026092808072424600_cit20","doi-asserted-by":"publisher","first-page":"2024.11.17.623896","DOI":"10.1101\/2024.11.17.623896","article-title":"Single Particle\nTracking of Genetically Encoded Nanoparticles: Optimizing Expression\nfor Cytoplasmic Diffusion Studies","author":"Korunova","year":"2024","journal-title":"bioRxiv"},{"issue":"18","key":"2026092808072424600_cit21","doi-asserted-by":"publisher","first-page":"188102","DOI":"10.1103\/PhysRevLett.100.188102","article-title":"Differential Dynamic\nMicroscopy: Probing Wave Vector\nDependent Dynamics with a Microscope","volume":"100","author":"Cerbino","year":"2008","journal-title":"Phys.\nRev. Lett."},{"issue":"11","key":"2026092808072424600_cit22","doi-asserted-by":"publisher","first-page":"110901","DOI":"10.1063\/1.5001027","article-title":"Perspective: Differential Dynamic Microscopy Extracts\nMulti-Scale Activity in Complex Fluids and Biological Systems","volume":"147","author":"Cerbino","year":"2017","journal-title":"J. Chem. Phys."},{"issue":"184","key":"2026092808072424600_cit23","doi-asserted-by":"publisher","first-page":"e63931","DOI":"10.3791\/63931","article-title":"Quantifying Cytoskeleton\nDynamics Using Differential Dynamic Microscopy","volume":"2022","author":"Verwei","year":"2022","journal-title":"J. Vis. Exp."},{"issue":"7","key":"2026092808072424600_cit24","doi-asserted-by":"publisher","first-page":"073804","DOI":"10.1103\/PhysRevMaterials.1.073804","article-title":"Differential\nDynamic Microscopy Microrheology of Soft Materials: A Tracking-Free\nDetermination of the Frequency-Dependent Loss and Storage Moduli","volume":"1","author":"Edera","year":"2017","journal-title":"Phys. Rev. Mater."},{"issue":"6","key":"2026092808072424600_cit25","doi-asserted-by":"publisher","first-page":"1074","DOI":"10.1038\/s41592-023-02149-7","article-title":"Temporal Analysis\nof Relative Distances (TARDIS) Is a Robust, Parameter-Free Alternative\nto Single-Particle Tracking","volume":"21","author":"Martens","year":"2024","journal-title":"Nat. Methods"},{"issue":"11","key":"2026092808072424600_cit26","doi-asserted-by":"publisher","first-page":"385","DOI":"10.3390\/biology9110385","article-title":"Cytoskeletal\nRemodeling in Cancer","volume":"9","author":"Aseervatham","year":"2020","journal-title":"Biology (Basel)."},{"issue":"8","key":"2026092808072424600_cit27","doi-asserted-by":"publisher","first-page":"1882","DOI":"10.3390\/cancers13081882","article-title":"Cytoskeletal Dynamics\nin Epithelial-Mesenchymal Transition: Insights into Therapeutic Targets\nfor Cancer Metastasis","volume":"13","author":"Datta","year":"2021","journal-title":"Cancers 2021, Vol. 13,\nPage 1882"},{"issue":"6","key":"2026092808072424600_cit28","doi-asserted-by":"publisher","first-page":"1727","DOI":"10.1039\/D4NA00003J","article-title":"Cytoplasmic Viscosity Is a Potential\nBiomarker for Metastatic Breast Cancer Cells","volume":"6","author":"Dessard","year":"2024","journal-title":"Nanoscale Adv."},{"issue":"6","key":"2026092808072424600_cit29","doi-asserted-by":"publisher","first-page":"869","DOI":"10.1002\/ijc.20195","article-title":"Genetic Evolution\nin Colon Cancer KM12 Cells and Metastatic Derivates","volume":"110","author":"Camps","year":"2004","journal-title":"Int. J. Cancer"},{"key":"2026092808072424600_cit30","doi-asserted-by":"publisher","first-page":"446","DOI":"10.1002\/1096-9896(2000)9999:9999&lt;::AID-PATH775&gt;3.0.CO;2-K","article-title":"Validation\nof a Model of Colon Cancer Progression","volume":"192","author":"Hewitt","year":"2000","journal-title":"J. Pathol."},{"key":"2026092808072424600_cit31","doi-asserted-by":"publisher","first-page":"101515","DOI":"10.1016\/j.mcpro.2026.101515","article-title":"Deciphering Stiffness-Driven Changes\nin Colorectal Cancer by Proteomics","volume":"25","author":"Cresens","year":"2026","journal-title":"Molecular\n& Cellular Proteomics"},{"key":"2026092808072424600_cit32","doi-asserted-by":"publisher","DOI":"10.1101\/2024.09.17.613503","article-title":"Synthesis and Mechanical Characterization\nof Polyacrylamide (PAAm) Hydrogels with Different Stiffnesses for\nLarge-Batch Cell Culture Applications","author":"Cresens","year":"2024","journal-title":"bioRxiv"},{"issue":"19","key":"2026092808072424600_cit330","doi-asserted-by":"publisher","first-page":"28656","DOI":"10.1364\/OE.401557","article-title":"Fast-Tracking of Single\nEmitters in Large Volumes with Nanometer Precision","volume":"28","author":"Louis","year":"2020","journal-title":"Optics Express"},{"key":"2026092808072424600_cit340","unstructured":"Huysecom, S.; Louis, B.\n          3D-image-processing Matlab code.\nhttps:\/\/github.com\/StevenHuysecom\/DDM-microrheology."},{"key":"2026092808072424600_cit33","doi-asserted-by":"publisher","DOI":"10.1101\/2021.11.18.469159","article-title":"NucGEMs Probe the\nBiophysical Properties of the Nucleoplasm","author":"Shu","year":"2022","journal-title":"bioRxiv"}],"container-title":["Chemical &amp; Biomedical\nImaging"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/cbmi.5c00261","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/cbihbp\/article-pdf\/4\/9\/1978\/66262169\/cbmi.5c00261.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/cbihbp\/article-pdf\/4\/9\/1978\/66262169\/cbmi.5c00261.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T12:33:07Z","timestamp":1790598787000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/cbihbp\/article\/4\/9\/1978\/5232864\/Quantitative-Analysis-of-Cytoplasmic-Viscosity-in"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,10]]},"references-count":35,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,4,10]]},"published-print":{"date-parts":[[2026,9,28]]}},"URL":"https:\/\/doi.org\/10.1021\/cbmi.5c00261","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2025.11.13.687967","asserted-by":"object"}]},"ISSN":["2832-3637"],"issn-type":[{"value":"2832-3637","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,10]]}}}