{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T14:45:53Z","timestamp":1773931553517,"version":"3.50.1"},"reference-count":61,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T00:00:00Z","timestamp":1773878400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T00:00:00Z","timestamp":1773878400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R01\u2010GM148644"],"award-info":[{"award-number":["R01\u2010GM148644"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["P01\u2010AI102851"],"award-info":[{"award-number":["P01\u2010AI102851"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytoskeleton"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>In this paper we describe a technique to make a confined environment of variable stiffness that is suitable for high\u2010resolution live\u2010cell imaging. This versatile and adaptable technique enables cell confinement between soft elastic surfaces made from polyacrylamide gels. The two surfaces retain all their compatibility with multiple approaches to chemically couple adhesion proteins, and additional techniques like micropatterning and traction force microscopy. This method is thus well suited for measuring force production and migration of weakly adherent cells that struggle to migrate in traditional planar environments.<\/jats:p>","DOI":"10.1002\/cm.70129","type":"journal-article","created":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T12:07:11Z","timestamp":1773922031000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Double Sided Traction Force Microscopy: A Method to Confine Cells for Physiologically Relevant Force Measurements"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-5752-6534","authenticated-orcid":false,"given":"Alexia","family":"Caillier","sequence":"first","affiliation":[{"name":"Department of Cell and Molecular Physiology, Stritch School of Medicine Loyola University Chicago  Maywood Illinois USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9951-1318","authenticated-orcid":false,"given":"Patrick W.","family":"Oakes","sequence":"additional","affiliation":[{"name":"Department of Cell and Molecular Physiology, Stritch School of Medicine Loyola University Chicago  Maywood Illinois USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2026,3,19]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"crossref","first-page":"109592","DOI":"10.1016\/j.ijmecsci.2024.109592","article-title":"Regularization Techniques and Inverse Approaches in 3D Traction Force Microscopy","volume":"283","author":"Apolinar\u2010Fern\u00e1ndez A.","year":"2024","journal-title":"International Journal of Mechanical Sciences"},{"key":"e_1_2_8_3_1","doi-asserted-by":"crossref","first-page":"15313","DOI":"10.1038\/ncomms15313","article-title":"Shifting the Optimal Stiffness for Cell Migration","volume":"8","author":"Bangasser B. L.","year":"2017","journal-title":"Nature Communications"},{"key":"e_1_2_8_4_1","doi-asserted-by":"crossref","first-page":"e1008364","DOI":"10.1371\/journal.pcbi.1008364","article-title":"pyTFM: A Tool for Traction Force and Monolayer Stress Microscopy","volume":"17","author":"Bauer A.","year":"2021","journal-title":"PLoS Computational Biology"},{"key":"e_1_2_8_5_1","doi-asserted-by":"crossref","first-page":"524","DOI":"10.1038\/ncb3134","article-title":"Force Transmission During Adhesion\u2010Independent Migration","volume":"17","author":"Bergert M.","year":"2015","journal-title":"Nature Cell Biology"},{"key":"e_1_2_8_6_1","doi-asserted-by":"crossref","first-page":"12814","DOI":"10.1038\/ncomms12814","article-title":"Confocal Reference Free Traction Force Microscopy","volume":"7","author":"Bergert M.","year":"2016","journal-title":"Nature Communications"},{"key":"e_1_2_8_7_1","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1152\/ajpcell.00270.2001","article-title":"Traction Fields, Moments, and Strain Energy That Cells Exert on Their Surroundings","volume":"282","author":"Butler J. P.","year":"2002","journal-title":"American Journal of Physiology"},{"key":"e_1_2_8_8_1","doi-asserted-by":"crossref","DOI":"10.1083\/jcb.202310067","article-title":"T Cells Use Focal Adhesions to Pull Themselves Through Confined Environments","volume":"223","author":"Caillier A.","year":"2024","journal-title":"Journal of Cell Biology"},{"key":"e_1_2_8_9_1","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1038\/nrm3897","article-title":"Physical Influences of the Extracellular Environment on Cell Migration","volume":"15","author":"Charras G.","year":"2014","journal-title":"Nature Reviews. Molecular Cell Biology"},{"key":"e_1_2_8_10_1","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1038\/s41580-022-00472-z","article-title":"Interplay Between Mechanics and Signalling in Regulating Cell Fate","volume":"23","author":"De Belly H.","year":"2022","journal-title":"Nature Reviews. Molecular Cell Biology"},{"key":"e_1_2_8_11_1","doi-asserted-by":"crossref","first-page":"e69850","DOI":"10.1371\/journal.pone.0069850","article-title":"Three\u2010Dimensional Quantification of Cellular Traction Forces and Mechanosensing of Thin Substrata by Fourier Traction Force Microscopy","volume":"8","author":"\u00c1lamo J. C.","year":"2013","journal-title":"PLoS One"},{"key":"e_1_2_8_12_1","doi-asserted-by":"crossref","unstructured":"Dembo M. andY.\u2010L.Wang.1999.Stresses at the Cell\u2010to\u2010Substrate Interface During Locomotion of Fibroblasts.","DOI":"10.1016\/S0006-3495(99)77386-8"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12195\u2010024\u201000801\u20106"},{"key":"e_1_2_8_14_1","doi-asserted-by":"crossref","first-page":"a005959","DOI":"10.1101\/cshperspect.a005959","article-title":"Signaling Networks That Regulate Cell Migration","volume":"7","author":"Devreotes P.","year":"2015","journal-title":"Cold Spring Harbor Perspectives in Biology"},{"key":"e_1_2_8_15_1","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1016\/j.cell.2006.06.044","article-title":"Matrix Elasticity Directs Stem Cell Lineage Specification","volume":"126","author":"Engler A. J.","year":"2006","journal-title":"Cell"},{"key":"e_1_2_8_16_1","doi-asserted-by":"crossref","first-page":"15799","DOI":"10.1021\/acsomega.5c02459","article-title":"Modulation of Gene Expression by Substrate Stiffness via Ubiquitination of Histone H2B by Ubiquitin\u2010Conjugating Enzyme E2A\/B","volume":"10","author":"Feng M.","year":"2025","journal-title":"ACS Omega"},{"key":"e_1_2_8_17_1","doi-asserted-by":"crossref","first-page":"e17833","DOI":"10.1371\/journal.pone.0017833","article-title":"Three\u2010Dimensional Traction Force Microscopy: A New Tool for Quantifying Cell\u2010Matrix Interactions","volume":"6","author":"Franck C.","year":"2011","journal-title":"PLoS One"},{"key":"e_1_2_8_18_1","doi-asserted-by":"crossref","first-page":"2990","DOI":"10.1016\/j.devcel.2024.06.023","article-title":"Rigidity Percolation and Active Advection Synergize in the Actomyosin Cortex to Drive Amoeboid Cell Motility","volume":"59","author":"Garc\u00eda\u2010Arcos J. M.","year":"2024","journal-title":"Developmental Cell"},{"key":"e_1_2_8_19_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fimmu.2016.00428","article-title":"T Cell Interstitial Migration: Motility Cues From the Inflamed Tissue for Micro\u2010 and Macro\u2010Positioning","volume":"7","author":"Gaylo A.","year":"2016","journal-title":"Frontiers in Immunology"},{"key":"e_1_2_8_20_1","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1038\/nmeth.3430","article-title":"Traction Microscopy to Identify Force Modulation in Subresolution Adhesions","volume":"12","author":"Han S. J.","year":"2015","journal-title":"Nature Methods"},{"key":"e_1_2_8_21_1","first-page":"177","article-title":"Silicone Rubber Substrata: A New Wrinkle in the Study of Cell Locomotion","volume":"208","author":"Harris A. K.","year":"1980","journal-title":"Science (1979)"},{"key":"e_1_2_8_22_1","doi-asserted-by":"crossref","first-page":"107313","DOI":"10.1016\/j.cpc.2020.107313","article-title":"A Bayesian Traction Force Microscopy Method With Automated Denoising in a User\u2010Friendly Software Package","volume":"256","author":"Huang Y.","year":"2020","journal-title":"Computer Physics Communications"},{"key":"e_1_2_8_23_1","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1038\/s41598-018-36896-x","article-title":"Traction Force Microscopy With Optimized Regularization and Automated Bayesian Parameter Selection for Comparing Cells","volume":"9","author":"Huang Y.","year":"2019","journal-title":"Scientific Reports"},{"key":"e_1_2_8_24_1","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1007\/s12195-009-0082-6","article-title":"Live Cells Exert 3\u2010Dimensional Traction Forces on Their Substrata","volume":"2","author":"Hur S. S.","year":"2009","journal-title":"Cellular and Molecular Bioengineering"},{"key":"e_1_2_8_25_1","doi-asserted-by":"crossref","first-page":"1081","DOI":"10.1038\/s41563-022-01294-2","article-title":"Directed Cell Migration Towards Softer Environments","volume":"21","author":"Isomursu A.","year":"2022","journal-title":"Nature Materials"},{"key":"e_1_2_8_26_1","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1152\/physrev.00013.2019","article-title":"Stiffness Sensing by Cells","volume":"100","author":"Janmey P. A.","year":"2020","journal-title":"Physiological Reviews"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.071001"},{"key":"e_1_2_8_28_1","doi-asserted-by":"crossref","first-page":"e1013079","DOI":"10.1371\/journal.pcbi.1013079","article-title":"Uncertainty\u2010Aware Traction Force Microscopy","volume":"21","author":"Kandasamy A.","year":"2025","journal-title":"PLoS Computational Biology"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2012.04.003"},{"key":"e_1_2_8_30_1","doi-asserted-by":"crossref","DOI":"10.3389\/fmats.2021.637278","article-title":"A Novel Method for Polyacrylamide Gel Preparation Using N\u2010Hydroxysuccinimide\u2010Acrylamide Ester to Study Cell\u2010Extracellular Matrix Mechanical Interactions","volume":"8","author":"Kumai J.","year":"2021","journal-title":"Frontiers in Materials"},{"key":"e_1_2_8_31_1","doi-asserted-by":"crossref","first-page":"9684","DOI":"10.1021\/acs.langmuir.5b01267","article-title":"Three\u2010Dimensional Confocal Microscopy Indentation Method for Hydrogel Elasticity Measurement","volume":"31","author":"Lee D.","year":"2015","journal-title":"Langmuir"},{"key":"e_1_2_8_32_1","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/j.biomaterials.2016.06.022","article-title":"N\u2010Terminal Specific Conjugation of Extracellular Matrix Proteins to 2\u2010Pyridinecarboxaldehyde Functionalized Polyacrylamide Hydrogels","volume":"102","author":"Lee J. P.","year":"2016","journal-title":"Biomaterials"},{"key":"e_1_2_8_33_1","doi-asserted-by":"crossref","first-page":"10097","DOI":"10.1073\/pnas.0900174106","article-title":"Microfabricated Tissue Gauges to Measure and Manipulate Forces From 3D Microtissues","volume":"106","author":"Legant W. R.","year":"2009","journal-title":"Proceedings of the National Academy of Sciences"},{"key":"e_1_2_8_34_1","doi-asserted-by":"crossref","first-page":"100090","DOI":"10.1016\/j.celbio.2025.100090","article-title":"Morphology\u2010Guided Cellular Behavior Modulation With 3D\u2010Printed Engineered ECM","volume":"1","author":"Liu N.","year":"2025","journal-title":"Cell Biomaterials"},{"key":"e_1_2_8_35_1","doi-asserted-by":"crossref","first-page":"659","DOI":"10.1016\/j.cell.2015.01.007","article-title":"Confinement and Low Adhesion Induce Fast Amoeboid Migration of Slow Mesenchymal Cells","volume":"160","author":"Liu Y. J.","year":"2015","journal-title":"Cell"},{"key":"e_1_2_8_36_1","doi-asserted-by":"crossref","first-page":"e08314","DOI":"10.7554\/eLife.08314","article-title":"Erk Regulation of Actin Capping and Bundling by Eps8 Promotes Cortex Tension and Leader Bleb\u2010Based Migration","volume":"4","author":"Logue J. S.","year":"2015","journal-title":"eLife"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrg3513"},{"key":"e_1_2_8_38_1","doi-asserted-by":"crossref","first-page":"3643","DOI":"10.1021\/acsami.1c17901","article-title":"Facile and Versatile Method for Micropatterning Poly(Acrylamide) Hydrogels Using Photocleavable Comonomers","volume":"14","author":"Missirlis D.","year":"2022","journal-title":"ACS Applied Materials & Interfaces"},{"key":"e_1_2_8_39_1","doi-asserted-by":"crossref","first-page":"4086","DOI":"10.1038\/s41598-019-40608-4","article-title":"Quantitative Reconstruction of Time\u2010Varying 3D Cell Forces With Traction Force Optical Coherence Microscopy","volume":"9","author":"Mulligan J. A.","year":"2019","journal-title":"Scientific Reports"},{"key":"e_1_2_8_40_1","first-page":"1","article-title":"The Physical and Biochemical Properties of the Extracellular Matrix Regulate Cell Fate","volume":"130","author":"Muncie J. M.","year":"2018","journal-title":"Current Topics in Developmental Biology"},{"key":"e_1_2_8_41_1","doi-asserted-by":"crossref","unstructured":"Munevar S. Y.\u2010L.Wang andM.Dembo.2001.Traction Force Microscopy of Migrating Normal and H\u2010Ras Transformed 3T3 Fibroblasts.","DOI":"10.1016\/S0006-3495(01)76145-0"},{"key":"e_1_2_8_42_1","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.ceb.2018.04.006","article-title":"Balancing Forces in Migration","volume":"54","author":"Oakes P. W.","year":"2018","journal-title":"Current Opinion in Cell Biology"},{"key":"e_1_2_8_43_1","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1016\/j.bpj.2014.06.045","article-title":"Geometry Regulates Traction Stresses in Adherent Cells","volume":"107","author":"Oakes P. W.","year":"2014","journal-title":"Biophysical Journal"},{"key":"e_1_2_8_44_1","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1101\/sqb.2016.81.030817","article-title":"Physical and Chemical Gradients in the Tumor Microenvironment Regulate Tumor Cell Invasion, Migration, and Metastasis","volume":"81","author":"Oudin M. J.","year":"2016","journal-title":"Cold Spring Harbor Symposia on Quantitative Biology"},{"key":"e_1_2_8_45_1","first-page":"13661","article-title":"Cell Locomotion and Focal Adhesions Are Regulated by Substrate Flexibility","volume":"94","author":"Pelham R. J.","year":"1997","journal-title":"Cell Biology"},{"key":"e_1_2_8_46_1","first-page":"367","article-title":"High\u2010Resolution Traction Force Microscopy","volume":"176","author":"Plotnikov S. V.","year":"2014","journal-title":"Physiology & Behavior"},{"key":"e_1_2_8_47_1","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1529\/biophysj.107.113670","article-title":"High Resolution Traction Force Microscopy Based on Experimental and Computational Advances","volume":"94","author":"Sabass B.","year":"2008","journal-title":"Biophysical Journal"},{"key":"e_1_2_8_48_1","doi-asserted-by":"crossref","first-page":"4426","DOI":"10.1529\/biophysj.108.132217","article-title":"Substrate Modulus Directs Neural Stem Cell Behavior","volume":"95","author":"Saha K.","year":"2008","journal-title":"Biophysical Journal"},{"key":"e_1_2_8_49_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.261338"},{"key":"e_1_2_8_50_1","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1016\/j.cell.2023.11.041","article-title":"Machine Learning Interpretable Models of Cell Mechanics From Protein Images","volume":"187","author":"Schmitt M. S.","year":"2024","journal-title":"Cell"},{"key":"e_1_2_8_51_1","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1016\/j.cub.2020.11.037","article-title":"Nuclear Deformation Causes DNA Damage by Increasing Replication Stress","volume":"31","author":"Shah P.","year":"2021","journal-title":"Current Biology"},{"key":"e_1_2_8_52_1","doi-asserted-by":"crossref","DOI":"10.1115\/1.4046974","article-title":"Recovery of Tractions Exerted by Single Cells in Three\u2010Dimensional Nonlinear Matrices","volume":"142","author":"Song D.","year":"2020","journal-title":"Journal of Biomechanical Engineering"},{"key":"e_1_2_8_53_1","doi-asserted-by":"crossref","first-page":"194104","DOI":"10.1088\/0953-8984\/22\/19\/194104","article-title":"Optimization of Traction Force Microscopy for Micron\u2010Sized Focal Adhesions","volume":"22","author":"Stricker J.","year":"2010","journal-title":"Journal of Physics. Condensed Matter"},{"key":"e_1_2_8_54_1","doi-asserted-by":"crossref","first-page":"e90976","DOI":"10.1371\/journal.pone.0090976","article-title":"High Resolution, Large Deformation 3D Traction Force Microscopy","volume":"9","author":"Toyjanova J.","year":"2014","journal-title":"PLoS One"},{"key":"e_1_2_8_55_1","doi-asserted-by":"crossref","DOI":"10.1038\/s41598-024-60971-1","article-title":"The Amoeboid Migration of Monocytes in Confining Channels Requires the Local Remodeling of the Cortical Actin Cytoskeleton by Cofilin\u20101","volume":"14","author":"Ullo M. F.","year":"2024","journal-title":"Scientific Reports"},{"key":"e_1_2_8_56_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2022.961460"},{"key":"e_1_2_8_57_1","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/B978-0-12-417136-7.00006-9","article-title":"Polyacrylamide Hydrogel Micropatterning","volume":"120","author":"Vignaud T.","year":"2014","journal-title":"Methods in Cell Biology"},{"key":"e_1_2_8_58_1","doi-asserted-by":"publisher","DOI":"10.3389\/fchem.2022.1012443"},{"key":"e_1_2_8_59_1","doi-asserted-by":"crossref","first-page":"7488","DOI":"10.1038\/s41598-024-56661-7","article-title":"The Impact of Substrate Stiffness on Morphological, Transcriptional and Functional Aspects in RPE","volume":"14","author":"Wolfram L.","year":"2024","journal-title":"Scientific Reports"},{"key":"e_1_2_8_60_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41580\u2010019\u20100172\u20109"},{"key":"e_1_2_8_61_1","doi-asserted-by":"publisher","DOI":"10.1016\/J.ENG.2017.01.014"},{"key":"e_1_2_8_62_1","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1002\/cm.20041","article-title":"Effects of Substrate Stiffness on Cell Morphology, Cytoskeletal Structure, and Adhesion","volume":"60","author":"Yeung T.","year":"2005","journal-title":"Cell Motility and the Cytoskeleton"}],"container-title":["Cytoskeleton"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cm.70129","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cm.70129","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cm.70129","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T12:07:17Z","timestamp":1773922037000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/cm.70129"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,19]]},"references-count":61,"alternative-id":["10.1002\/cm.70129"],"URL":"https:\/\/doi.org\/10.1002\/cm.70129","archive":["Portico"],"relation":{},"ISSN":["1949-3584","1949-3592"],"issn-type":[{"value":"1949-3584","type":"print"},{"value":"1949-3592","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,19]]},"assertion":[{"value":"2025-08-26","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-03-02","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-03-19","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70129"}}