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Sensors 2011, 11(6), 6214-6236; doi:10.3390/s110606214
Article

Interfacial Chemistry and the Design of Solid-Phase Nucleic Acid Hybridization Assays Using Immobilized Quantum Dots as Donors in Fluorescence Resonance Energy Transfer

 and *
Received: 26 April 2011; in revised form: 26 May 2011 / Accepted: 7 June 2011 / Published: 9 June 2011
(This article belongs to the Special Issue Sensing with Quantum Dots)
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Abstract: The use of quantum dots (QDs) as donors in fluorescence resonance energy transfer (FRET) offer several advantages for the development of multiplexed solid-phase QD-FRET nucleic acid hybridization assays. Designs for multiplexing have been demonstrated, but important challenges remain in the optimization of these systems. In this work, we identify several strategies based on the design of interfacial chemistry for improving sensitivity, obtaining lower limits of detection (LOD) and enabling the regeneration and reuse of solid-phase QD-FRET hybridization assays. FRET-sensitized emission from acceptor dyes associated with hybridization events at immobilized QD donors provides the analytical signal in these assays. The minimization of active sensing area reduces background from QD donor PL and allows the resolution of smaller amounts of acceptor emission, thus lowering the LOD. The association of multiple acceptor dyes with each hybridization event can enhance FRET efficiency, thereby improving sensitivity. Many previous studies have used interfacial protein layers to generate selectivity; however, transient destabilization of these layers is shown to prevent efficient regeneration. To this end, we report a protein-free interfacial chemistry and demonstrate the specific detection of as little as 2 pmol of target, as well as an improved capacity for regeneration.
Keywords: active area; biosensor; DNA; fluorescence resonance energy transfer; immobilization; regeneration; signal enhancement; quantum dots active area; biosensor; DNA; fluorescence resonance energy transfer; immobilization; regeneration; signal enhancement; quantum dots
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Algar, W.R.; Krull, U.J. Interfacial Chemistry and the Design of Solid-Phase Nucleic Acid Hybridization Assays Using Immobilized Quantum Dots as Donors in Fluorescence Resonance Energy Transfer. Sensors 2011, 11, 6214-6236.

AMA Style

Algar WR, Krull UJ. Interfacial Chemistry and the Design of Solid-Phase Nucleic Acid Hybridization Assays Using Immobilized Quantum Dots as Donors in Fluorescence Resonance Energy Transfer. Sensors. 2011; 11(6):6214-6236.

Chicago/Turabian Style

Algar, W. Russ; Krull, Ulrich J. 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, no. 6: 6214-6236.


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