{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T18:24:19Z","timestamp":1791224659718,"version":"4.1.0"},"reference-count":36,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2012,2,9]],"date-time":"2012-02-09T00:00:00Z","timestamp":1328745600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>We have developed a novel microarray technology based on total internal reflection fluorescence (TIRF) in combination with DNA and protein bioassays immobilized at the TIRF surface. Unlike conventional microarrays that exhibit reduced signal-to-background ratio, require several stages of incubation, rinsing and stringency control, and measure only end-point results, our TIRF microarray technology provides several orders of magnitude better signal-to-background ratio, performs analysis rapidly in one step, and measures the entire course of association and dissociation kinetics between target DNA and protein molecules and the bioassays. In many practical cases detection of only DNA or protein markers alone does not provide the necessary accuracy for diagnosing a disease or detecting a pathogen. Here we describe TIRF microarrays that detect DNA and protein markers simultaneously, which reduces the probabilities of false responses. Supersensitive and multiplexed TIRF DNA and protein microarray technology may provide a platform for accurate diagnosis or enhanced research studies. Our TIRF microarray system can be mounted on upright or inverted microscopes or interfaced directly with CCD cameras equipped with a single objective, facilitating the development of portable devices. As proof-of-concept we applied TIRF microarrays for detecting molecular markers from Bacillus anthracis, the pathogen responsible for anthrax.<\/jats:p>","DOI":"10.3390\/s120201800","type":"journal-article","created":{"date-parts":[[2012,2,9]],"date-time":"2012-02-09T11:22:30Z","timestamp":1328786550000},"page":"1800-1815","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence"],"prefix":"10.3390","volume":"12","author":[{"given":"Alexander","family":"Asanov","sequence":"first","affiliation":[{"name":"TIRF Technologies, 951 Aviation Parkway, Suite 700, Morrisville, NC 27560, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ang\u00e9lica","family":"Zepeda","sequence":"additional","affiliation":[{"name":"Instituto de Investigaciones Biom\u00e9dicas, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Ciudad Universitaria, DF 04510, M\u00e9xico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis","family":"Vaca","sequence":"additional","affiliation":[{"name":"Instituto de Fisiolog\u00eda Celular, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Ciudad Universitaria, DF 04510, M\u00e9xico"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2012,2,9]]},"reference":[{"key":"ref_1","first-page":"368","article-title":"New developments in fluorescence diagnostics","volume":"9","author":"Hegyi","year":"2011","journal-title":"J. Dtsch. Dermatol. Ges"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2685","DOI":"10.1021\/cr900267p","article-title":"Fluorescence polarization\/anisotropy in diagnostics and imaging","volume":"110","author":"Jameson","year":"2010","journal-title":"Chem. Rev"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"11753","DOI":"10.1073\/pnas.94.22.11753","article-title":"Fluorescence correlation spectroscopy: Diagnostics for sparse molecules","volume":"94","author":"Maiti","year":"1997","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1088\/0031-9155\/42\/5\/006","article-title":"In vivo fluorescence imaging for tissue diagnostics","volume":"42","author":"Klinteberg","year":"1997","journal-title":"Phys. Med. Biol"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1007\/s00216-010-4276-5","article-title":"Recent developments and applications of hybrid surface plasmon resonance interfaces in optical sensing","volume":"399","author":"Gao","year":"2011","journal-title":"Anal. Bioanal. Chem"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1016\/j.aca.2010.12.022","article-title":"A review of monolithic multichannel quartz crystal microbalance: A review","volume":"687","author":"Tuantranont","year":"2011","journal-title":"Anal. Chim. Acta"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"4427","DOI":"10.1002\/adma.201000740","article-title":"Functional organic field-effect transistors","volume":"22","author":"Guo","year":"2010","journal-title":"Adv. Mater"},{"key":"ref_8","first-page":"389","article-title":"Fluorescence-based thermal shift assays","volume":"13","author":"Zhang","year":"2010","journal-title":"Curr. Opin. Drug Discov. Devel"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/S0079-6468(05)43002-7","article-title":"Fluorescence-based assays","volume":"43","author":"Eccleston","year":"2005","journal-title":"Prog. Med. Chem"},{"key":"ref_10","first-page":"S435","article-title":"Surface-enhanced Raman scattering biomedical applications of plasmonic colloidal particles","volume":"7","year":"2010","journal-title":"J. R. Soc. Interface"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1007\/s00216-009-2682-3","article-title":"Surface-enhanced Raman scattering (SERS) for probing internal cellular structure and dynamics","volume":"394","author":"Willets","year":"2009","journal-title":"Anal. Bioanal. Chem"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"955","DOI":"10.1039\/b708841h","article-title":"Practical understanding and use of surface enhanced Raman scattering\/surface enhanced resonance Raman scattering in chemical and biological analysis","volume":"37","author":"Smith","year":"2008","journal-title":"Chem. Soc. Rev"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.tibtech.2008.09.004","article-title":"Applications of microarrays in pathogen detection and biodefense","volume":"27","author":"Uttamchandani","year":"2009","journal-title":"Trends Biotechnol"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"659","DOI":"10.1373\/clinchem.2008.112821","article-title":"Microarray-based genomic DNA profiling technologies in clinical molecular diagnostics","volume":"55","author":"Shen","year":"2009","journal-title":"Clin. Chem"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1097\/MOP.0b013e328320d87e","article-title":"Molecular diagnostics of infectious diseases","volume":"21","author":"Muldrew","year":"2009","journal-title":"Curr. Opin. Pediatr"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1007\/978-1-60761-670-2_12","article-title":"Preparation of lipid membrane surfaces for molecular interaction studies by surface plasmon resonance biosensors","volume":"627","author":"Besenicar","year":"2010","journal-title":"Methods Mol. Biol"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"928","DOI":"10.1016\/j.jprot.2009.01.027","article-title":"Design of high-density antibody microarrays for disease proteomics: Key technological issues","volume":"72","author":"Borrebaeck","year":"2009","journal-title":"J. Proteomics"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2143","DOI":"10.2174\/0929867043364739","article-title":"High density DNA microarrays: Algorithms and biomedical applications","volume":"11","author":"Liu","year":"2004","journal-title":"Curr. Med. Chem"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1930","DOI":"10.3390\/ijms10041930","article-title":"Fluorescence lifetime imaging of quantum dot labeled DNA microarrays","volume":"10","author":"Giraud","year":"2009","journal-title":"Int. J. Mol. Sci"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1023\/B:MODI.0000036233.58271.25","article-title":"High density peptide microarrays. In situ synthesis and applications","volume":"8","author":"Gao","year":"2004","journal-title":"Mol. Divers"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1007\/s00216-006-0317-5","article-title":"Optical technologies for the read out and quality control of DNA and protein microarrays","volume":"385","author":"Schaferling","year":"2006","journal-title":"Anal. Bioanal. Chem"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Fish, K.N. (2009). Total internal reflection fluorescence (TIRF) microscopy. Curr. Protoc. Cytom.","DOI":"10.1002\/0471142956.cy1218s50"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/j.bbalip.2009.10.004","article-title":"A novel form of Total Internal Reflection Fluorescence Microscopy (LG-TIRFM) reveals different and independent lipid raft domains in living cells","volume":"1801","author":"Asanov","year":"2010","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Gu, X., Qiu, T., Zhang, W., and Chu, P.K. (2011). Light-emitting diodes enhanced by localized surface plasmon resonance. Nanoscale Res. Lett, 6.","DOI":"10.1186\/1556-276X-6-199"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1038\/nbt0396-303","article-title":"Molecular beacons: Probes that fluoresce upon hybridization","volume":"14","author":"Tyagi","year":"1996","journal-title":"Nat. Biotechnol"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1016\/j.mib.2005.08.001","article-title":"Point-of-care molecular diagnostic systems\u2014Past, present and future","volume":"8","author":"Holland","year":"2005","journal-title":"Curr. Opin. Microbiol"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1126\/science.270.5235.467","article-title":"Quantitative monitoring of gene expression patterns with a complementary DNA microarray","volume":"270","author":"Schena","year":"1995","journal-title":"Science"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1016\/S0956-5663(03)00021-6","article-title":"High-density, microsphere-based fiber optic DNA microarrays","volume":"18","author":"Epstein","year":"2003","journal-title":"Biosens. Bioelectron"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.mad.2006.11.021","article-title":"Protein microarray technology","volume":"128","author":"Hall","year":"2007","journal-title":"Mech. Ageing Dev"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1101\/gr.4235806","article-title":"Development of a microscopic platform for real-time monitoring of biomolecular interactions","volume":"16","author":"Sasuga","year":"2006","journal-title":"Genome Res"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.bioeng.2006.03.002","article-title":"Merging microfluidics with microarray-based bioassays","volume":"23","author":"Situma","year":"2006","journal-title":"Biomol. Eng"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"124","DOI":"10.2174\/187220808784619711","article-title":"Diagnosis of pathogens using DNA microarray","volume":"2","author":"Yoo","year":"2008","journal-title":"Recent Pat. Biotechnol"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1007\/978-0-387-39978-2_11","article-title":"SNP and mutation analysis","volume":"593","author":"Wang","year":"2007","journal-title":"Adv. Exp. Med. Biol"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2411","DOI":"10.1007\/s00216-008-2121-x","article-title":"Photochemical and analytical applications of gold nanoparticles and nanorods utilizing surface plasmon resonance","volume":"391","author":"Uechi","year":"2008","journal-title":"Anal. Bioanal. Chem"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1067","DOI":"10.1016\/j.micinf.2008.07.003","article-title":"Do DNA microarrays have their future behind them?","volume":"10","author":"Coppee","year":"2008","journal-title":"Microbes. Infect"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"8079","DOI":"10.1021\/ac1013157","article-title":"In-gel technology for PCR genotyping and pathogen detection","volume":"82","author":"Atrazhev","year":"2010","journal-title":"Anal. Chem"}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/12\/2\/1800\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:48:49Z","timestamp":1760219329000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/12\/2\/1800"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,2,9]]},"references-count":36,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2012,2]]}},"alternative-id":["s120201800"],"URL":"https:\/\/doi.org\/10.3390\/s120201800","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,2,9]]}}}