Immobilization of CdSe/ZnS quantum dots on glass beads for the detection of nucleic acid hybridization using fluorescence resonance energy transfer

W. Russ Algar; Ulrich J. Krull

doi:10.1117/12.878344

11 February 2011 Immobilization of CdSe/ZnS quantum dots on glass beads for the detection of nucleic acid hybridization using fluorescence resonance energy transfer

W. Russ Algar, Ulrich J. Krull

Author Affiliations +

Proceedings Volume 7909, Colloidal Quantum Dots/Nanocrystals for Biomedical Applications VI; 79090C (2011) https://doi.org/10.1117/12.878344
Event: SPIE BiOS, 2011, San Francisco, California, United States

Abstract

The photoluminescence (PL) properties of quantum dots (QD) are of significant interest in the development of new methods for bioanalysis. Multiplexed solid-phase nucleic acid hybridization assays that use immobilized QDs as donors in fluorescence resonance energy transfer (FRET) are one such example, and offer several unique advantages over other methods. In this work, new interfacial chemistry is described for the immobilization of red-emitting CdSe/ZnS QDs on glass beads for use in hybridization assays. The beads were chemically modified with a dithiolate surface ligand and the QDs immobilized via self-assembly. Further derivatization of the QDs with dithiolate-terminated probe oligonucleotides enabled a hybridization assay that could detect unlabeled target down to nanomolar levels with discrimination of single base-pair mismatches. The use of beads as an immobilization platform afforded shorter analysis times and superior reusability compared to previous studies using optical fibers. Hybridization between probe, target, and Alexa Fluor 647 (A647) labeled reporter oligonucleotides in a sandwich format generated a spectroscopic signal by introducing the proximity needed for FRET between the QDs and A647. The results indicate clear directions for the optimization of solid-phase hybridization assays, and are important for the future development of true multiplexed biosensors based on QDs and FRET.

Citation Download Citation

W. Russ Algar and Ulrich J. Krull "Immobilization of CdSe/ZnS quantum dots on glass beads for the detection of nucleic acid hybridization using fluorescence resonance energy transfer", Proc. SPIE 7909, Colloidal Quantum Dots/Nanocrystals for Biomedical Applications VI, 79090C (11 February 2011); https://doi.org/10.1117/12.878344

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