Toward an on-chip multiplexed nucleic acid hybridization assay using immobilized quantum dot-oligonucleotide conjugates and fluorescence resonance energy transfer

Anthony J. Tavares; M. Omair Noor; W. Russ Algar; Charles H. Vannoy; Lu Chen; Ulrich J. Krull

doi:10.1117/12.877326

11 February 2011 Toward an on-chip multiplexed nucleic acid hybridization assay using immobilized quantum dot-oligonucleotide conjugates and fluorescence resonance energy transfer

Anthony J. Tavares, M. Omair Noor, W. Russ Algar, Charles H. Vannoy, Lu Chen, Ulrich J. Krull

Author Affiliations +

Proceedings Volume 7909, Colloidal Quantum Dots/Nanocrystals for Biomedical Applications VI; 79090X (2011) https://doi.org/10.1117/12.877326
Event: SPIE BiOS, 2011, San Francisco, California, United States

Abstract

Semiconductor quantum dots (QD) are a class of NP with photophysical properties that are ideally suited for optical multiplexing and use as donors in fluorescence resonance energy transfer (FRET). A new strategy is presented for the development of multiplexed DNA hybridization assays using immobilized QDs in a microfluidic system. Green- or red-emitting QDs were immobilized via self-assembly with a multidentate-thiol-derivatized glass slide, and subsequently conjugated with amine-terminated probe oligonucleotides using carbodiimide activation. Immobilized QD-probe conjugates were then passivated with adsorbed non-complementary oligonucleotides to achieve selectivity in microfluidic assays. Target nucleic acid sequences hybridized with QD-probe conjugates and were labeled with Cy3 or Alexa Fluor 647 as acceptor dyes for the QD donors, where FRET-sensitized dye emission provided a signal for the detection of picomolar quantities of target. The simultaneous immobilization of green- and red-emitting QDs at different ratios within a microfluidic channel was demonstrated as a step toward multiplexed assays.

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Anthony J. Tavares, M. Omair Noor, W. Russ Algar, Charles H. Vannoy, Lu Chen, and Ulrich J. Krull "Toward an on-chip multiplexed nucleic acid hybridization assay using immobilized quantum dot-oligonucleotide conjugates and fluorescence resonance energy transfer", Proc. SPIE 7909, Colloidal Quantum Dots/Nanocrystals for Biomedical Applications VI, 79090X (11 February 2011); https://doi.org/10.1117/12.877326

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