{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,12]],"date-time":"2025-11-12T13:51:48Z","timestamp":1762955508865,"version":"build-2065373602"},"reference-count":122,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2016,2,15]],"date-time":"2016-02-15T00:00:00Z","timestamp":1455494400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Micromachines"],"abstract":"<jats:p>Optical detection has long been the most popular technique in immunosensing. Recent developments in the synthesis of luminescent probes and the fabrication of novel nanostructures enable more sensitive and efficient optical detection, which can be miniaturized and integrated with microfluidics to realize compact lab-on-a-chip immunosensors. These immunosensors are portable, economical and automated, but their sensitivity is not compromised. This review focuses on the incorporation and implementation of optical detection and microfluidics in immunosensors; it introduces the working principles of each optical detection technique and how it can be exploited in immunosensing. The recent progress in various opto-microfluidic immunosensor designs is described. Instead of being comprehensive to include all opto-microfluidic platforms, the report centers on the designs that are promising for point-of-care immunosensing diagnostics, in which ease of use, stability and cost-effective fabrication are emphasized.<\/jats:p>","DOI":"10.3390\/mi7020029","type":"journal-article","created":{"date-parts":[[2016,2,15]],"date-time":"2016-02-15T06:23:07Z","timestamp":1455517387000},"page":"29","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Opto-Microfluidic Immunosensors: From Colorimetric to Plasmonic"],"prefix":"10.3390","volume":"7","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4301-0829","authenticated-orcid":false,"given":"Jie-Long","family":"He","sequence":"first","affiliation":[{"name":"Department of Mechanical Engineering, National Taiwan University, Taipei 10617, Taiwan"}]},{"given":"Da-Shin","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering, National Taiwan University, Taipei 10617, Taiwan"}]},{"given":"Shih-Kang","family":"Fan","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering, National Taiwan University, Taipei 10617, Taiwan"}]}],"member":"1968","published-online":{"date-parts":[[2016,2,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1016\/0731-7085(94)80027-8","article-title":"Chemiluminescence and immunoassays","volume":"12","author":"Rongen","year":"1994","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1042\/BA20070019","article-title":"Dendrimers and their applications in immunoassays and clinical diagnostics","volume":"48","author":"Singh","year":"2007","journal-title":"Biotechnol. Appl. Biochem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"749","DOI":"10.1586\/14737159.6.5.749","article-title":"Signal amplification systems in immunoassays: Implications for clinical diagnostics","volume":"6","author":"Dhawan","year":"2006","journal-title":"Expert Rev. Mol. Diagn."},{"key":"ref_4","first-page":"1","article-title":"Immunoassay techniques","volume":"324","author":"Wheeler","year":"2006","journal-title":"Methods Mol. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1038\/358603a0","article-title":"Automated real-time immunoassay of biomolecules","volume":"358","author":"Afeyan","year":"1992","journal-title":"Nature"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/j.vetmic.2009.09.064","article-title":"Detection of H6 influenza antibody by blocking enzyme-linked immunosorbent assay","volume":"142","author":"Chen","year":"2010","journal-title":"Vet. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.jim.2012.09.009","article-title":"Preparation of monoclonal antibodies against poor immunogenic avian influenza virus proteins","volume":"387","author":"He","year":"2013","journal-title":"J. Immunol. Methods"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1016\/j.vetmic.2014.10.008","article-title":"A monoclonal antibody recognizes a highly conserved neutralizing epitope on hemagglutinin of H6N1 avian influenza virus","volume":"174","author":"He","year":"2014","journal-title":"Vet. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3422","DOI":"10.1039\/c2an16294f","article-title":"Detection of influenza virus using a lateral flow immunoassay for amplified DNA by a microfluidic RT-PCR chip","volume":"137","author":"Nagatani","year":"2012","journal-title":"Analyst"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1006\/jmbi.1996.0548","article-title":"Antibody-antigen interactions: Contact analysis and binding site topography","volume":"262","author":"MacCallum","year":"1996","journal-title":"J. Mol. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/0022-1759(92)90061-W","article-title":"Enzyme immunoassay techniques an overview","volume":"150","author":"Porstmann","year":"1992","journal-title":"J. Immunol. Methods"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2488","DOI":"10.1016\/j.bios.2004.10.016","article-title":"Microfluidic immunosensor systems","volume":"20","author":"Bange","year":"2005","journal-title":"Biosens. Bioelectron."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1007\/s10404-010-0638-8","article-title":"Microfluidic-based biosensors toward point-of-care detection of nucleic acids and proteins","volume":"10","author":"Choi","year":"2011","journal-title":"Microfluid. Nanofluid."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.aca.2008.01.064","article-title":"Current development in microfluidic immunosensing chip","volume":"611","author":"Henares","year":"2008","journal-title":"Anal. Chim. Acta"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/j.jala.2010.01.013","article-title":"Microfluidic immunoassays","volume":"15","author":"Lin","year":"2010","journal-title":"J. Lab. Autom."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1016\/j.tibtech.2005.07.007","article-title":"Artificial, non-antibody binding proteins for pharmaceutical and industrial applications","volume":"23","author":"Hey","year":"2005","journal-title":"Trends Biotechnol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1628","DOI":"10.1093\/clinchem\/45.9.1628","article-title":"Aptamers: An emerging class of molecules that rival antibodies in diagnostics","volume":"45","author":"Jayasena","year":"1999","journal-title":"Clin. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1016\/j.bios.2013.06.060","article-title":"Site-directed antibody immobilization techniques for immunosensors","volume":"50","author":"Makaraviciute","year":"2013","journal-title":"Biosens. Bioelectron."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/S0003-2697(02)00442-6","article-title":"Optimizing antibody immobilization strategies for the construction of protein microarrays","volume":"312","author":"Peluso","year":"2003","journal-title":"Anal. Biochem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4431","DOI":"10.1039\/c1an15325k","article-title":"Effect of antibody immobilization strategies on the analytical performance of a surface plasmon resonance-based immunoassay","volume":"136","author":"Vashist","year":"2011","journal-title":"Analyst"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/j.aca.2003.01.001","article-title":"Probing direct binding affinity in electrochemical antibody-based sensors","volume":"496","author":"Corry","year":"2003","journal-title":"Anal. Chim. Acta"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1887","DOI":"10.1016\/j.bios.2005.10.027","article-title":"Electrochemical biosensors: Towards point-of-care cancer diagnostics","volume":"21","author":"Wang","year":"2006","journal-title":"Biosens. Bioelectron."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4683","DOI":"10.1039\/c2cc16958d","article-title":"Electrochemical immunoassay on a 3D microfluidic paper-based device","volume":"48","author":"Zang","year":"2012","journal-title":"Chem. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"5012","DOI":"10.1021\/ac200926e","article-title":"Development of an optical biosensor based immunoassay to screen infant formula milk samples for adulteration with melamine","volume":"83","author":"Fodey","year":"2011","journal-title":"Anal. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1016\/j.talanta.2005.09.049","article-title":"Determination of environmental organic pollutants with a portable optical immunosensor","volume":"69","author":"Mauriz","year":"2006","journal-title":"Talanta"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1007\/s00604-012-0831-4","article-title":"Electromechanical biosensors for pathogen detection","volume":"178","author":"Xu","year":"2012","journal-title":"Microchim. Acta"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1006\/abio.1993.1345","article-title":"Enzymatic amplification of a flow-injected thermometric enzyme-linked immunoassay for human insulin","volume":"212","author":"Mecklenburg","year":"1993","journal-title":"Anal. Biochem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4063","DOI":"10.1039\/c1lc20362b","article-title":"Microspot-based ELISA in microfluidics: Chemiluminescence and colorimetry detection using integrated thin-film hydrogenated amorphous silicon photodiodes","volume":"11","author":"Novo","year":"2011","journal-title":"Lab Chip"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1243","DOI":"10.1039\/b816018j","article-title":"Flow-through functionalized PDMS microfluidic channels with dextran derivative for ELISAs","volume":"9","author":"Yu","year":"2009","journal-title":"Lab Chip"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1021\/ac8020592","article-title":"Aptamer-functionalized gold nanoparticles as probes in a dry-reagent strip biosensor for protein analysis","volume":"81","author":"Xu","year":"2009","journal-title":"Anal. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1548","DOI":"10.1039\/b820321k","article-title":"A fully automated immunoassay from whole blood on a disc","volume":"9","author":"Lee","year":"2009","journal-title":"Lab Chip"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"030504","DOI":"10.1117\/1.JBO.19.3.030504","article-title":"Ultrasensitive opto-microfluidic immunosensor integrating gold nanoparticle-enhanced chemiluminescence and highly stable organic photodetector","volume":"19","author":"Pires","year":"2014","journal-title":"J. Biomed. Opt."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Yao, P., Liu, Z., Tung, S., Dong, Z., and Liu, L. (2015). Fully automated quantification of insulin concentration using a microfluidic-based chemiluminescence immunoassay. J. Lab. Autom.","DOI":"10.1177\/2211068215578822"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3150","DOI":"10.1039\/c2lc40325k","article-title":"3D origami-based multifunction-integrated immunodevice: Low-cost and multiplexed sandwich chemiluminescence immunoassay on microfluidic paper-based analytical device","volume":"12","author":"Ge","year":"2012","journal-title":"Lab Chip"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.talanta.2014.06.075","article-title":"Automated chemiluminescence immunoassay for a nonionic surfactant using a recycled spinning-pausing controlled washing procedure on a compact disc-type microfluidic platform","volume":"133","author":"Guo","year":"2015","journal-title":"Talanta"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3776","DOI":"10.1039\/C5LC00660K","article-title":"A digital microfluidic device with integrated nanostructured microelectrodes for electrochemical immunoassays","volume":"15","author":"Rackus","year":"2015","journal-title":"Lab Chip"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4489","DOI":"10.1039\/c2lc40707h","article-title":"Battery-triggered microfluidic paper-based multiplex electrochemiluminescence immunodevice based on potential-resolution strategy","volume":"12","author":"Wang","year":"2012","journal-title":"Lab Chip"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"4207","DOI":"10.1021\/ac300551e","article-title":"Microchip device with 64-site electrode array for multiplexed immunoassay of cell surface antigens based on electrochemiluminescence resonance energy transfer","volume":"84","author":"Wu","year":"2012","journal-title":"Anal. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/j.bios.2015.08.050","article-title":"Fluorescence enhancement and multiple protein detection in ZnO nanostructure microfluidic devices","volume":"75","author":"Sang","year":"2016","journal-title":"Biosens. Bioelectron."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.bios.2015.05.041","article-title":"High sensitivity automated multiplexed immunoassays using photonic crystal enhanced fluorescence microfluidic system","volume":"73","author":"Tan","year":"2015","journal-title":"Biosens. Bioelectron."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1007\/s10544-014-9881-1","article-title":"A nanostructured aluminum oxide-based microfluidic device for enhancing immunoassay\u2019s fluorescence and detection sensitivity","volume":"16","author":"Li","year":"2014","journal-title":"Biomed. Microdevices"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1021\/ac980798t","article-title":"An array immunosensor for simultaneous detection of clinical analytes","volume":"71","author":"Rowe","year":"1999","journal-title":"Anal. Chem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2091","DOI":"10.1039\/b814922d","article-title":"Development of a digital microfluidic platform for point of care testing","volume":"8","author":"Sista","year":"2008","journal-title":"Lab Chip"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1039\/b920589f","article-title":"Parallel microfluidic surface plasmon resonance imaging arrays","volume":"10","author":"Ouellet","year":"2010","journal-title":"Lab Chip"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2218","DOI":"10.1016\/j.bios.2008.11.031","article-title":"Biochip functionalization using electrowetting-on-dielectric digital microfluidics for surface plasmon resonance imaging detection of DNA hybridization","volume":"24","author":"Malic","year":"2009","journal-title":"Biosens. Bioelectron."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1039\/B814697G","article-title":"Two-dimensional droplet-based surface plasmon resonance imaging using electrowetting-on-dielectric microfluidics","volume":"9","author":"Malic","year":"2009","journal-title":"Lab Chip"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.aca.2007.08.046","article-title":"Optical sensing systems for microfluidic devices: A review","volume":"601","author":"Kuswandi","year":"2007","journal-title":"Anal. Chim. Acta"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1016\/0005-2795(71)90132-2","article-title":"Enzyme-linked immunosorbent assay. II. Quantitative assay of protein antigen, immunoglobulin G, by means of enzyme-labelled antigen and antibody-coated tubes","volume":"251","author":"Engvall","year":"1971","journal-title":"Biochim. Biophys. Acta Protein Struct."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2415","DOI":"10.1373\/clinchem.2005.051532","article-title":"Enzyme immunoassay (EIA)\/enzyme-linked immunosorbent assay (ELISA)","volume":"51","author":"Lequin","year":"2005","journal-title":"Clin. Chem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1275","DOI":"10.1021\/ac802228k","article-title":"User configurable microfluidic device for multiplexed immunoassays based on DNA-directed assembly","volume":"81","author":"Schroeder","year":"2009","journal-title":"Anal. Chem."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"601","DOI":"10.1146\/annurev.bioeng.8.061505.095758","article-title":"Lab on a CD","volume":"8","author":"Madou","year":"2006","journal-title":"Annu. Rev. Biomed. Eng."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1016\/S0039-9140(99)00294-5","article-title":"Chemiluminescence as diagnostic tool. A review","volume":"51","author":"Dodeigne","year":"2000","journal-title":"Talanta"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"404","DOI":"10.1016\/j.trac.2008.12.006","article-title":"Chemiluminescence immunoassay","volume":"28","author":"Zhao","year":"2009","journal-title":"Trends Anal. Chem."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"587","DOI":"10.2116\/analsci.25.587","article-title":"Chemiluminescence platforms in immunoassay and DNA analyses","volume":"25","author":"Fan","year":"2009","journal-title":"Anal. Sci."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2994","DOI":"10.1021\/ac015645b","article-title":"Microfluidic enzyme immunoassay using silicon microchip with immobilized antibodies and chemiluminescence detection","volume":"74","author":"Yakovleva","year":"2002","journal-title":"Anal. Chem."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1016\/S0731-7085(98)00062-4","article-title":"Chemiluminescence-based detection: Principles and analytical applications in flowing streams and in immunoassays","volume":"17","author":"Baeyens","year":"1998","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1042\/bj2900051","article-title":"Peroxynitrite-induced luminol chemiluminescence","volume":"290","author":"Radi","year":"1993","journal-title":"Biochem. J."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"5484","DOI":"10.1021\/ac900724m","article-title":"Automated support-resolution strategy for a one-way chemiluminescent multiplex immunoassay","volume":"81","author":"Yang","year":"2009","journal-title":"Anal. Chem."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1368","DOI":"10.1016\/j.bios.2006.06.003","article-title":"Automated 10-channel capillary chip immunodetector for biological agents detection","volume":"22","author":"Hell","year":"2007","journal-title":"Biosens. Bioelectron."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"238","DOI":"10.1016\/j.bios.2011.12.021","article-title":"Paper-based three-dimensional electrochemical immunodevice based on multi-walled carbon nanotubes functionalized paper for sensitive point-of-care testing","volume":"32","author":"Wang","year":"2012","journal-title":"Biosens. Bioelectron."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1042\/cs0700403","article-title":"Chemiluminescence immunoassay: An overview","volume":"70","author":"Weeks","year":"1986","journal-title":"Clin. Sci."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"2714","DOI":"10.1039\/c3lc50170a","article-title":"Chemiluminescence detector based on a single planar transparent digital microfluidic device","volume":"13","author":"Zeng","year":"2013","journal-title":"Lab Chip"},{"key":"ref_63","unstructured":"Chemical Connection. Available online: http:\/\/www.chemicalconnection.org.uk\/chemistry\/topics\/images\/ee7b.jpg."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"3275","DOI":"10.1039\/b923679c","article-title":"Applications and trends in electrochemiluminescence","volume":"39","author":"Hu","year":"2010","journal-title":"Chem. Soc. Rev."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"3003","DOI":"10.1021\/cr020373d","article-title":"Electrochemiluminescence (ECL)","volume":"104","author":"Richter","year":"2004","journal-title":"Chem. Rev."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1146\/annurev-anchem-060908-155305","article-title":"Electrogenerated chemiluminescence","volume":"2","author":"Forster","year":"2009","journal-title":"Annu. Rev. Anal. Chem."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"2655","DOI":"10.1110\/ps.021860","article-title":"Construction of a fluorescent biosensor family","volume":"11","author":"Smith","year":"2002","journal-title":"Protein Sci."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1021\/bc015584o","article-title":"Peptide arrays for highly sensitive and specific antibody-binding fluorescence assays","volume":"13","author":"Melnyk","year":"2002","journal-title":"Bioconjug. Chem."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/j.bios.2014.02.012","article-title":"In-situ fluorescent immunomagnetic multiplex detection of foodborne pathogens in very low numbers","volume":"57","author":"Cho","year":"2014","journal-title":"Biosens. Bioelectron."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.autrev.2012.02.013","article-title":"Antinuclear antibody detection by automated multiplex immunoassay in untreated patients at the time of diagnosis","volume":"12","author":"Vermeersch","year":"2012","journal-title":"Autoimmun. Rev."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.cbpa.2013.11.012","article-title":"Cross-reactivity in antibody microarrays and multiplexed sandwich assays: Shedding light on the dark side of multiplexing","volume":"18","author":"Juncker","year":"2014","journal-title":"Curr. Opin. Chem. Biol."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.aca.2012.04.044","article-title":"Development of an immunomagnetic bead-based time-resolved fluorescence immunoassay for rapid determination of levels of carcinoembryonic antigen in human serum","volume":"734","author":"Hou","year":"2012","journal-title":"Anal. Chim. Acta"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"15083","DOI":"10.1021\/ja306066a","article-title":"Amine-functionalized lanthanide-doped zirconia nanoparticles: Optical spectroscopy, time-resolved fluorescence resonance energy transfer biodetection, and targeted imaging","volume":"134","author":"Liu","year":"2012","journal-title":"J. Am. Chem. Soc."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"8461","DOI":"10.1021\/ic400384f","article-title":"Stable and highly fluorescent europium (III) chelates for time-resolved immunoassays","volume":"52","author":"Wang","year":"2013","journal-title":"Inorg. Chem."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1016\/S0956-5663(02)00077-5","article-title":"A label-free optical technique for detecting small molecule interactions","volume":"17","author":"Lin","year":"2002","journal-title":"Biosens. Bioelectron."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.aca.2008.05.022","article-title":"Sensitive optical biosensors for unlabeled targets: A review","volume":"620","author":"Fan","year":"2008","journal-title":"Anal. Chim. Acta"},{"key":"ref_77","unstructured":"Bennett, C.A. (2008). Principles of Physical Optics, Wiley."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"2750","DOI":"10.1016\/j.tsf.2010.12.175","article-title":"Development of biosensor based on imaging ellipsometry and biomedical applications","volume":"519","author":"Jin","year":"2011","journal-title":"Thin Solid Films"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"3301","DOI":"10.1021\/nn900825q","article-title":"A label-free porous alumina interferometric immunosensor","volume":"3","author":"Alvarez","year":"2009","journal-title":"ACS Nano"},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"2070","DOI":"10.1021\/nn901312f","article-title":"A stable, label-free optical interferometric biosensor based on TiO2 nanotube arrays","volume":"4","author":"Mun","year":"2010","journal-title":"ACS Nano"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/S0925-4005(98)00321-9","article-title":"Surface plasmon resonance sensors: Review","volume":"54","author":"Homola","year":"1999","journal-title":"Sens. Actuators B Chem."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1385\/ABAB:126:2:079","article-title":"Surface plasmon resonance: Applications in understanding receptor-ligand interaction","volume":"126","author":"Pattnaik","year":"2005","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"7257","DOI":"10.1021\/ac049318q","article-title":"Label-free reading of microarray-based immunoassays with surface plasmon resonance imaging","volume":"76","author":"Kanda","year":"2004","journal-title":"Anal. Chem."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1146\/annurev.physchem.58.032806.104607","article-title":"Localized surface plasmon resonance spectroscopy and sensing","volume":"58","author":"Willets","year":"2007","journal-title":"Annu. Rev. Phys. Chem."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"1624","DOI":"10.1021\/ac011127p","article-title":"Development of an aggregation-based immunoassay for anti-protein a using gold nanoparticles","volume":"74","author":"Thanh","year":"2002","journal-title":"Anal. Chem."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1007\/s00216-008-2287-2","article-title":"Lateral flow (immuno) assay: Its strengths, weaknesses, opportunities and threats. A literature survey","volume":"393","author":"Korf","year":"2009","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"1119","DOI":"10.1039\/c2lc20751f","article-title":"Progress toward multiplexed sample-to-result detection in low resource settings using microfluidic immunoassay cards","volume":"12","author":"Lafleur","year":"2012","journal-title":"Lab Chip"},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"4574","DOI":"10.1021\/ac300689s","article-title":"Two-dimensional paper network format that enables simple multistep assays for use in low-resource settings in the context of malaria antigen detection","volume":"84","author":"Fu","year":"2012","journal-title":"Anal. Chem."},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"991","DOI":"10.1007\/s00216-010-3678-8","article-title":"Immunoassays in microfluidic systems","volume":"397","author":"Ng","year":"2010","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"3249","DOI":"10.1039\/c2lc40630f","article-title":"Microfluidic designs and techniques using lab-on-a-chip devices for pathogen detection for point-of-care diagnostics","volume":"12","author":"Foudeh","year":"2012","journal-title":"Lab Chip"},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"1988","DOI":"10.1039\/b814043j","article-title":"Lab on paper","volume":"8","author":"Zhao","year":"2008","journal-title":"Lab Chip"},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"4771","DOI":"10.1002\/anie.201001005","article-title":"Paper-based ELISA","volume":"49","author":"Cheng","year":"2010","journal-title":"Angew. Chem. Int. Ed. Engl."},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"19606","DOI":"10.1073\/pnas.0810903105","article-title":"Three-dimensional microfluidic devices fabricated in layered paper and tape","volume":"105","author":"Martinez","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"17564","DOI":"10.1021\/ja2071779","article-title":"Three-dimensional paper microfluidic devices assembled using the principles of origami","volume":"133","author":"Liu","year":"2011","journal-title":"J. Am. Chem. Soc."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1016\/j.biomaterials.2011.10.065","article-title":"Three-dimensional paper-based electrochemiluminescence immunodevice for multiplexed measurement of biomarkers and point-of-care testing","volume":"33","author":"Ge","year":"2012","journal-title":"Biomaterials"},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"5140","DOI":"10.1021\/ac300397h","article-title":"Eco-friendly plasmonic sensors: Using the photothermal effect to prepare metal nanoparticle-containing test papers for highly sensitive colorimetric detection","volume":"84","author":"Tseng","year":"2012","journal-title":"Anal. Chem."},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"1236","DOI":"10.1039\/b816535a","article-title":"General digital microfluidic platform manipulating dielectric and conductive droplets by dielectrophoresis and electrowetting","volume":"9","author":"Fan","year":"2009","journal-title":"Lab Chip"},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"2500","DOI":"10.1039\/c1lc20142e","article-title":"Encapsulated droplets with metered and removable oil shells by electrowetting and dielectrophoresis","volume":"11","author":"Fan","year":"2011","journal-title":"Lab Chip"},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1039\/C4LC00794H","article-title":"A droplet-to-digital (D2D) microfluidic device for single cell assays","volume":"15","author":"Shih","year":"2015","journal-title":"Lab Chip"},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"3739","DOI":"10.1039\/C4LC00592A","article-title":"An EWOD-based microfluidic chip for single-cell isolation, mRNA purification and subsequent multiplex qPCR","volume":"14","author":"Rival","year":"2014","journal-title":"Lab Chip"},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"1325","DOI":"10.1039\/b803204a","article-title":"Cross-scale electric manipulations of cells and droplets by frequency-modulated dielectrophoresis and electrowetting","volume":"8","author":"Fan","year":"2008","journal-title":"Lab Chip"},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"1330","DOI":"10.1039\/b704084a","article-title":"Asymmetric electrowetting\u2014Moving droplets by a square wave","volume":"7","author":"Fan","year":"2007","journal-title":"Lab Chip"},{"key":"ref_103","doi-asserted-by":"crossref","first-page":"4833","DOI":"10.1021\/ac0498112","article-title":"Electrowetting-based microfluidics for analysis of peptides and proteins by matrix-assisted laser desorption\/ionization mass spectrometry","volume":"76","author":"Wheeler","year":"2004","journal-title":"Anal. Chem."},{"key":"ref_104","doi-asserted-by":"crossref","first-page":"6121","DOI":"10.1021\/ac5012969","article-title":"Analysis on the go: Quantitation of drugs of abuse in dried urine with digital microfluidics and miniature mass spectrometry","volume":"86","author":"Kirby","year":"2014","journal-title":"Anal. Chem."},{"key":"ref_105","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1021\/ac048754+","article-title":"Digital microfluidics with in-line sample purification for proteomics analyses with MALDI-MS","volume":"77","author":"Wheeler","year":"2005","journal-title":"Anal. Chem."},{"key":"ref_106","doi-asserted-by":"crossref","first-page":"8466","DOI":"10.1021\/ac5022198","article-title":"Digital microfluidic platform for human plasma protein depletion","volume":"86","author":"Mei","year":"2014","journal-title":"Anal. Chem."},{"key":"ref_107","doi-asserted-by":"crossref","first-page":"6474","DOI":"10.1021\/ac103230c","article-title":"Evaluation of digital PCR for absolute DNA quantification","volume":"83","author":"Sanders","year":"2011","journal-title":"Anal Chem"},{"key":"ref_108","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.jviromet.2011.09.017","article-title":"Digital PCR provides absolute quantitation of viral load for an occult RNA virus","volume":"179","author":"White","year":"2012","journal-title":"J. Virol. Methods"},{"key":"ref_109","doi-asserted-by":"crossref","first-page":"22319","DOI":"10.3390\/ijms160922319","article-title":"Digital microfluidics for manipulation and analysis of a single cell","volume":"16","author":"He","year":"2015","journal-title":"Int. J. Mol. Sci."},{"key":"ref_110","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1039\/b717759c","article-title":"Digital microfluidics for cell-based assays","volume":"8","author":"Yang","year":"2008","journal-title":"Lab Chip"},{"key":"ref_111","doi-asserted-by":"crossref","first-page":"2210","DOI":"10.1039\/c3lc50169h","article-title":"Paper-based microfluidic point-of-care diagnostic devices","volume":"13","author":"Yetisen","year":"2013","journal-title":"Lab Chip"},{"key":"ref_112","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1586\/erm.11.22","article-title":"Applications of electrowetting-based digital microfluidics in clinical diagnostics","volume":"11","author":"Pollack","year":"2011","journal-title":"Expert Rev. Mol. Diagn."},{"key":"ref_113","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1146\/annurev-anchem-062011-143028","article-title":"Digital microfluidics","volume":"5","author":"Choi","year":"2012","journal-title":"Annu. Rev. Anal. Chem."},{"key":"ref_114","doi-asserted-by":"crossref","first-page":"2188","DOI":"10.1039\/b807855f","article-title":"Heterogeneous immunoassays using magnetic beads on a digital microfluidic platform","volume":"8","author":"Sista","year":"2008","journal-title":"Lab Chip"},{"key":"ref_115","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1007\/s00216-010-4368-2","article-title":"A digital microfluidic approach to heterogeneous immunoassays","volume":"399","author":"Miller","year":"2011","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_116","doi-asserted-by":"crossref","first-page":"054026","DOI":"10.1088\/0960-1317\/21\/5\/054026","article-title":"A versatile electrowetting-based digital microfluidic platform for quantitative homogeneous and heterogeneous bio-assays","volume":"21","author":"Vergauwe","year":"2011","journal-title":"J. Micromech. Microeng."},{"key":"ref_117","doi-asserted-by":"crossref","first-page":"8805","DOI":"10.1021\/ac3020627","article-title":"Digital microfluidic magnetic separation for particle-based immunoassays","volume":"84","author":"Ng","year":"2012","journal-title":"Anal. Chem."},{"key":"ref_118","doi-asserted-by":"crossref","first-page":"1384","DOI":"10.1016\/j.talanta.2011.03.073","article-title":"Electrochemical detection on electrowetting-on-dielectric digital microfluidic chip","volume":"84","author":"Karuwan","year":"2011","journal-title":"Talanta"},{"key":"ref_119","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1039\/C3LC51063H","article-title":"A digital microfluidic electrochemical immunoassay","volume":"14","author":"Shamsi","year":"2014","journal-title":"Lab Chip"},{"key":"ref_120","doi-asserted-by":"crossref","first-page":"5967","DOI":"10.1007\/s00216-014-7997-z","article-title":"A fluorogenic heterogeneous immunoassay for cardiac muscle troponin cTnI on a digital microfluidic device","volume":"406","author":"Tsaloglou","year":"2014","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_121","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1373\/clinchem.2014.232181","article-title":"Digital microfluidic platform for the detection of rubella infection and immunity: A proof of concept","volume":"61","author":"Ng","year":"2015","journal-title":"Clin. Chem."},{"key":"ref_122","doi-asserted-by":"crossref","first-page":"9638","DOI":"10.1021\/ac401847x","article-title":"Automated digital microfluidic platform for magnetic-particle-based immunoassays with optimization by design of experiments","volume":"85","author":"Choi","year":"2013","journal-title":"Anal. Chem."}],"container-title":["Micromachines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-666X\/7\/2\/29\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T19:19:06Z","timestamp":1760210346000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-666X\/7\/2\/29"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,2,15]]},"references-count":122,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2016,2]]}},"alternative-id":["mi7020029"],"URL":"https:\/\/doi.org\/10.3390\/mi7020029","relation":{},"ISSN":["2072-666X"],"issn-type":[{"type":"electronic","value":"2072-666X"}],"subject":[],"published":{"date-parts":[[2016,2,15]]}}}