Abstract
Semiconductor quantum dots (QDs) have long served as integral components in signal transduction modalities such as Förster resonance energy transfer (FRET). The majority of bioanalytical methods using QDs for FRET-based techniques simply monitor binding-induced conformational changes. In more recent work, QDs have been incorporated into solid-phase support systems, such as microfluidic chips, to serve as physical platforms in the development of functional biosensors and bioprobes. Herein, we describe a simple strategy for the transduction of nucleic acid hybridization that combines a novel design method based on FRET with an electrokinetically controlled microfluidic technology, and that offers further potential for amelioration of sample-handling issues and for simplification of dynamic stringency control.
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References
Peter M (2001) Microfluidics – downsizing large-scale biology. Nat Biotechnol 19:717–721
Meldrum DR, Holl MR (2002) Microscale bioanalytical systems. Science 297:1197–1198
Sia SK, Whitesides GM (2003) Microfluidic devices fabricated in poly(dimethylsiloxane) for biological studies. Electrophoresis 24:3563–3576
Whitesides GM (2006) The origins and the future of microfluidics. Nature 442:368–373
Erickson D, Li D (2004) Integrated microfluidic devices. Anal Chim Acta 507:11–26
Vannoy CH, Tavares AJ, Noor MO, Uddayasankar U, Krull UJ (2011) Biosensing with quantum dots: a microfluidic approach. Sensors 11:9732–9763
Tavares AJ, Noor MO, Vannoy CH, Algar WR, Krull UJ (2012) On-chip transduction of nucleic acid hybridization using spatial profiles of immobilized quantum dots and fluorescence resonance energy transfer. Anal Chem 84:312–319
Algar WR, Krull UJ (2009) Interfacial transduction of nucleic acid hybridization using immobilized quantum dots as donors in fluorescence resonance energy transfer. Langmuir 25:633–638
Algar WR, Krull UJ (2009) Toward a multiplexed solid-phase nucleic acid hybridization assay using quantum dots as donors in fluorescence resonance energy transfer. Anal Chem 81:4113–4120
Algar WR, Krull UJ (2010) Developing mixed films of immobilized oligonucleotides and quantum dots for the multiplexed detection of nucleic acid hybridization using a combination of fluorescence resonance energy transfer and direct excitation of fluorescence. Langmuir 26:6041–6047
Algar WR, Krull UJ (2011) Interfacial chemistry and the design of solid-phase nucleic acid hybridization assays using immobilized quantum dots as donors in fluorescence resonance energy transfer. Sensors 11:6214–6236
Petryayeva E, Algar WR, Krull UJ (2013) Adapting fluorescence resonance energy transfer with quantum dot donors for solid-phase hybridization assays in microtiter plate format. Langmuir 29:977–987
Noor MO, Shahmuradyan A, Krull UJ (2013) Paper-based solid-phase nucleic acid hybridization assay using immobilized quantum dots as donors in fluorescence resonance energy transfer. Anal Chem 85:1860–1867
Chen L, Algar WR, Tavares AJ, Krull UJ (2011) Toward a solid-phase nucleic acid hybridization assay within microfluidic channels using immobilized quantum dots as donors in fluorescence resonance energy transfer. Anal Bioanal Chem 399:133–141
Susumu K, Mei BC, Mattoussi H (2009) Multifunctional ligands based on dihydrolipoic acid and polyethylene glycol to promote biocompatibility of quantum dots. Nat Protoc 4:424–436
Erickson D, Sinton D, Li D (2003) Joule heating and heat transfer in poly(dimethylsiloxane) microfluidic systems. Lab Chip 3:141–149
Clapp AR, Medintz IL, Mattoussi H (2006) Forster resonance energy transfer investigations using quantum dot fluorophores. ChemPhysChem 7:47–57
Vannoy CH, Chong L, Le C, Krull UJ (2013) A competitive displacement assay with quantum dots as fluorescence resonance energy transfer donors. Anal Chim Acta 759:92–99
Petryayeva E, Krull UJ (2012) Quantum dot and gold nanoparticle immobilization for biosensing applications using multidentate imidazole surface ligands. Langmuir 28:13943–13951
Noor MO, Krull UJ (2011) Microfluidics for the deposition of density gradients of immobilized oligonucleotide probes; developing surfaces that offer spatial control of the stringency of DNA hybridization. Anal Chim Acta 708:1–10
McDonald JC, Whitesides GM (2002) Poly(dimethylsiloxane) as a material for fabricating microfluidic devices. Acc Chem Res 35:491–499
Acknowledgements
The authors gratefully acknowledge financial support of their research program by the Natural Sciences and Engineering Research Council of Canada (NSERC). A.J.T. and U.U. are also thankful to NSERC for provision of graduate fellowships. M.O.N. is grateful to the Ontario Ministry of Training, Colleges and Universities (MTCU) for provision of an Ontario Graduate Scholarship (OGS).
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Tavares, A.J., Noor, M.O., Uddayasankar, U., Krull, U.J., Vannoy, C.H. (2014). Solid-Phase Supports for the in situ Assembly of Quantum Dot-FRET Hybridization Assays in Channel Microfluidics. In: Fontes, A., Santos, B. (eds) Quantum Dots: Applications in Biology. Methods in Molecular Biology, vol 1199. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1280-3_18
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DOI: https://doi.org/10.1007/978-1-4939-1280-3_18
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