{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,13]],"date-time":"2025-11-13T01:56:54Z","timestamp":1762999014106,"version":"build-2065373602"},"reference-count":48,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2014,2,6]],"date-time":"2014-02-06T00:00:00Z","timestamp":1391644800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Micromachines"],"abstract":"<jats:p>This review describes the use of ultrasound for inducing and retaining cell-cell contact in multi-well microplates combined with live-cell fluorescence microscopy. This platform has been used for studying the interaction between natural killer (NK) cells and cancer cells at the level of individual cells. The review includes basic principles of ultrasonic particle manipulation, design criteria when building a multi-well microplate device for this purpose, biocompatibility aspects, and finally, two examples of biological applications: Dynamic imaging of the inhibitory immune synapse, and studies of the heterogeneity in killing dynamics of NK cells interacting with cancer cells.<\/jats:p>","DOI":"10.3390\/mi5010027","type":"journal-article","created":{"date-parts":[[2014,2,6]],"date-time":"2014-02-06T11:48:42Z","timestamp":1391687322000},"page":"27-49","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":27,"title":["Ultrasound-Induced Cell\u2013Cell Interaction Studies in a Multi-Well Microplate"],"prefix":"10.3390","volume":"5","author":[{"given":"Martin","family":"Wiklund","sequence":"first","affiliation":[{"name":"Department of Applied Physics, KTH\u2014Royal Institute of Technology, SE-106 91 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Athanasia","family":"Christakou","sequence":"additional","affiliation":[{"name":"Department of Applied Physics, KTH\u2014Royal Institute of Technology, SE-106 91 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mathias","family":"Ohlin","sequence":"additional","affiliation":[{"name":"Department of Applied Physics, KTH\u2014Royal Institute of Technology, SE-106 91 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ida","family":"Iranmanesh","sequence":"additional","affiliation":[{"name":"Department of Applied Physics, KTH\u2014Royal Institute of Technology, SE-106 91 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas","family":"Frisk","sequence":"additional","affiliation":[{"name":"Department of Applied Physics, KTH\u2014Royal Institute of Technology, SE-106 91 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bruno","family":"Vanherberghen","sequence":"additional","affiliation":[{"name":"Department of Applied Physics, KTH\u2014Royal Institute of Technology, SE-106 91 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bj\u00f6rn","family":"\u00d6nfelt","sequence":"additional","affiliation":[{"name":"Department of Applied Physics, KTH\u2014Royal Institute of Technology, SE-106 91 Stockholm, Sweden"},{"name":"Department of Microbiology, Tumor and Cell Biology, Karolinska Institute, SE-171 21 Stockholm, Sweden"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2014,2,6]]},"reference":[{"key":"ref_1","unstructured":"Lindstr\u00f6m, S., and Andersson-Svahn, H. (2012). Methods in Molecular Biology, Humana Press Inc."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1038\/27094","article-title":"Calcium\u2014A life and death signal","volume":"395","author":"Berridge","year":"1998","journal-title":"Nature"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1126\/science.2683075","article-title":"G1 events and regulation of cell proliferation","volume":"246","author":"Pardee","year":"1989","journal-title":"Science"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1111\/j.1600-065X.1975.tb00164.x","article-title":"Functional analysis of B cell heterogeneity","volume":"24","author":"Ronowicz","year":"1975","journal-title":"Immunol. Rev."},{"key":"ref_5","first-page":"641","article-title":"Flow cytometry and cell sorting of heterogeneous microbial populations: The importance of single-cell analyses","volume":"60","author":"Davey","year":"1996","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3221","DOI":"10.1039\/c2lc40757d","article-title":"Acoustofluidics 18: Microscopy for acoustofluidic micro-devices","volume":"12","author":"Wiklund","year":"2012","journal-title":"Lab Chip"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2018","DOI":"10.1039\/c2lc40201g","article-title":"Acoustofluidics 12: Biocompatibility and cell viability in microfluidic acoustic resonators","volume":"12","author":"Wiklund","year":"2012","journal-title":"Lab Chip"},{"key":"ref_8","unstructured":"Crowther, J.R. (2001). Methods in Molecular Biology, Humana Press Inc."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"300","DOI":"10.3389\/fimmu.2012.00300","article-title":"Novel microchip-based tools facilitating live cell imaging and assessment of functional heterogeneity within NK cell populations","volume":"3","author":"Forslund","year":"2012","journal-title":"Front. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1219","DOI":"10.1002\/elps.200700536","article-title":"Towards high-throughput single cell\/clone cultivation and analysis","volume":"29","author":"Larsson","year":"2008","journal-title":"Electrophoresis"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e15453","DOI":"10.1371\/journal.pone.0015453","article-title":"Imaging immune surveillance of individual natural killer cells confined in microwell arrays","volume":"5","author":"Guldevall","year":"2010","journal-title":"PLoS One"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"683","DOI":"10.1007\/s10544-011-9538-2","article-title":"A silicon-glass microwell platform for high-resolution imaging and high-content screening with single cell resolution","volume":"13","author":"Frisk","year":"2011","journal-title":"Biomed. Microdevices"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1326","DOI":"10.1182\/blood-2012-06-439851","article-title":"Classification of human natural killer cells based on migration behavior and cytotoxic response","volume":"121","author":"Vanherberghen","year":"2013","journal-title":"Blood"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1039\/c2ib20167d","article-title":"Single-cell analysis of the dynamics and functional outcomes of interactions between human natural killer cells and target cells","volume":"4","author":"Yamanaka","year":"2012","journal-title":"Integr. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"770","DOI":"10.1039\/c1ib00007a","article-title":"Analysis of transient migration behavior of natural killer cells imaged in situ and in vitro","volume":"3","author":"Khorshidi","year":"2011","journal-title":"Integr. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2727","DOI":"10.1039\/c004707d","article-title":"Ultrasound-controlled cell aggregation in a multi-well chip","volume":"10","author":"Vanherberghen","year":"2010","journal-title":"Lab Chip"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"035008","DOI":"10.1088\/0960-1317\/23\/3\/035008","article-title":"Influence of acoustic streaming on ultrasonic particle manipulation in a 100-well ring-transducer microplate","volume":"23","author":"Ohlin","year":"2013","journal-title":"J. Micromech. Microeng."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"712","DOI":"10.1039\/c3ib20253d","article-title":"Live cell imaging in a micro-array of acoustic traps facilitates quantification of natural killer cell heterogeneity","volume":"5","author":"Christakou","year":"2013","journal-title":"Integr. Biol"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Lord Rayleigh (1905). On the momentum and pressure of gaseous vibrations, and on the connexion with virial theorem. Philos. Mag., 10, 364\u2013374.","DOI":"10.1080\/14786440509463381"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1121\/1.381833","article-title":"Radiation pressure\u2014The history of a mislabeled tensor","volume":"63","author":"Beyer","year":"1978","journal-title":"J. Acoust. Soc. Am."},{"key":"ref_21","first-page":"773","article-title":"On the forces acting on a small particle in an acoustical field in an ideal fluid","volume":"6","year":"1962","journal-title":"Sov. Phys. Dokl."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"095025","DOI":"10.1088\/0960-1317\/18\/9\/095025","article-title":"Wedge transducer design for two-dimensional ultrasonic manipulation in a microfluidic chip","volume":"18","author":"Manneberg","year":"2008","journal-title":"J. Micromech. Microeng."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/0301-5629(89)90157-9","article-title":"Theoretical criterion for acoustic aggregation","volume":"15","author":"Nyborg","year":"1989","journal-title":"Ultrasound Med. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"063901","DOI":"10.1063\/1.2971030","article-title":"A three-dimensional ultrasonic cage for characterization of individual cells","volume":"93","author":"Manneberg","year":"2008","journal-title":"Appl. Phys. Lett."},{"key":"ref_25","unstructured":"Barnkob, R., Augustsson, P., Magnusson, C., Lilja, H., Laurell, T., and Bruus, H. (2011, January 2\u20136). Measuring Density and Compressibility of White Blood Cells and Prostate Cancer Cells by Microchannel Acoustophoresis. Proceedings of the 15th MicroTAS, Seattle, WA, USA."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"4072","DOI":"10.1039\/c1lc20687g","article-title":"On-chip measurements of cell compressibility via acoustic radiation","volume":"11","author":"Hartono","year":"2011","journal-title":"Lab Chip"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1063\/1.3703290","article-title":"Efficient finite element modeling of acoustic radiation forces on inhomogeneous elastic particles","volume":"1433","author":"Mishra","year":"2012","journal-title":"AIP Conf. Proc."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"105002","DOI":"10.1088\/0960-1317\/23\/10\/105002","article-title":"Tunable-angle wedge transducer for improved acoustophoretic control in a microfluidic chip","volume":"23","author":"Iranmanesh","year":"2013","journal-title":"J. Micromech. Microeng."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1039\/C1LC20770A","article-title":"Acoustofluidics 2: Perturbation theory and ultrasound resonance modes","volume":"12","author":"Bruus","year":"2012","journal-title":"Lab Chip"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2438","DOI":"10.1039\/c2lc40203c","article-title":"Acoustofluidics 14: Applications of acoustic streaming in microfluidic devices","volume":"12","author":"Wiklund","year":"2012","journal-title":"Lab Chip"},{"key":"ref_31","unstructured":"Ohlin, M., Christakou, A.E., Frisk, T., \u00d6nfelt, B., and Wiklund, M. (2011, January 2\u20136). Controlling Acoustic Streaming in a Multi-well Microplate for Improving Live Cell Assays. Proceedings of the 15th International Conference on Miniaturized Systems for Chemistry and Life, Sciences (MicroTAS 2011), Seattle, WA, USA."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2337","DOI":"10.1039\/c2lc40120g","article-title":"Measuring acoustic energy density in microchannel acoustophoresis using a simple and rapid light-intensity method","volume":"12","author":"Barnkob","year":"2012","journal-title":"Lab Chip"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1039\/b920376a","article-title":"Measuring the local pressure amplitude in microchannel acoustophoresis","volume":"10","author":"Barnkob","year":"2010","journal-title":"Lab Chip"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/j.ultrasmedbio.2006.07.024","article-title":"Proliferation and viability of adherent cells manipulated by standing-wave ultrasound in a microfluidic chip","volume":"33","author":"Manneberg","year":"2007","journal-title":"Ultrasound Med. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Raffel, M., Willert, C.E., Wereley, S.T., and Kompenhans, J. (2007). Particle Image Velocimetry, Springer.","DOI":"10.1007\/978-3-540-72308-0"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"4152","DOI":"10.1039\/c1lc20637k","article-title":"Automated and temperature-controlled micro-PIV measurements enabling long-term-stable microchannel acoustophoresis characterization","volume":"11","author":"Augustsson","year":"2011","journal-title":"Lab Chip"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"056307","DOI":"10.1103\/PhysRevE.86.056307","article-title":"Acoustic radiation- and streaming-induced microparticle velocities determined by microparticle image velocimetry in an ultrasound symmetry plane","volume":"86","author":"Barnkob","year":"2013","journal-title":"Phys. Rev. E"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/0301-5629(91)90125-G","article-title":"Gauging the likelihood of cavitation from short-pulse, low-duty cycle diagnostic ultrasound","volume":"17","author":"Apfel","year":"1991","journal-title":"Ultrasound Med. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"7954","DOI":"10.1021\/ac301723s","article-title":"Microfluidic, label-free enrichment of prostate cancer cells in blood based on acoustophoresis","volume":"84","author":"Augustsson","year":"2012","journal-title":"Anal. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e64233","DOI":"10.1371\/journal.pone.0064233","article-title":"Microchannel acoustophoresis does not impact survival or function of microglia, leukocytes or tumor cells","volume":"8","author":"Burguillos","year":"2013","journal-title":"PLoS One"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1007\/s10565-009-9123-0","article-title":"Biochemical properties of encapsulated high-density 3-D HepG2 aggregates formed in an ultrasound trap for application in hepatotoxicity studies","volume":"26","author":"Bazou","year":"2010","journal-title":"Cell Biol. Toxicol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1038\/nri2381","article-title":"Formation and function of the lytic NK-cell immunological synapse","volume":"8","author":"Orange","year":"2008","journal-title":"Nat. Rev. Immunol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1038\/25764","article-title":"Three-dimensional segregation of supramolecular activation clusters in T cells","volume":"395","author":"Monks","year":"1998","journal-title":"Nature"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1126\/science.285.5425.221","article-title":"The immunological synapse: A molecular machine controlling T cell activation","volume":"285","author":"Grakoui","year":"1999","journal-title":"Science"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/0167-5699(90)90097-S","article-title":"In search of the \u201cmissing self\u201d: MHC molecules and NK cell recognition","volume":"11","author":"Ljunggren","year":"1990","journal-title":"Immunol. Today"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"e326","DOI":"10.1371\/journal.pone.0000326","article-title":"Serial killing of tumor cells by human natural killer cells\u2014Enhancement by therapeutic antibodies","volume":"2","author":"Bhat","year":"2007","journal-title":"PLoS One"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2369","DOI":"10.1182\/blood-2008-03-143727","article-title":"MHC class I-specific inhibitory receptors and their ligands structure diverse human NK-cell repertoires toward a balance of missing self-response","volume":"112","author":"Yawata","year":"2008","journal-title":"Blood"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"2434","DOI":"10.1182\/blood-2008-05-156836","article-title":"The strength of inhibitory input during education quantitatively tunes the functional responsiveness of individual natural killer cells","volume":"113","author":"Brodin","year":"2009","journal-title":"Blood"}],"container-title":["Micromachines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-666X\/5\/1\/27\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:07:58Z","timestamp":1760216878000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-666X\/5\/1\/27"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,2,6]]},"references-count":48,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2014,3]]}},"alternative-id":["mi5010027"],"URL":"https:\/\/doi.org\/10.3390\/mi5010027","relation":{},"ISSN":["2072-666X"],"issn-type":[{"type":"electronic","value":"2072-666X"}],"subject":[],"published":{"date-parts":[[2014,2,6]]}}}