The unique optical properties of quantum dots (QDs) have been widely used to develop bioassays based on Fluorescence Resonance Energy Transfer (FRET). The solid-phase assays using QDs as FRET donors have numerous practical advantages, including at least 10-fold enhancement in FRET efficiency, which is not immediately explained by theoretical predictions that model energy transfer processes of QDs in two-dimensional layers. Donor-acceptor separation distance, acceptor and donor concentrations were found to influence FRET efficiency in solid-phase assays. A novel immobilization strategy was implemented which made use of the high affinity of imidazole moieties to QD shells to build solid-phase QD bioassays. A 96-well polystyrene plate is presented as a platform suitable for rapid and convenient multiplexed detection. A typical microtiter plate reader was shown to be capable of discriminating different FRET pairs to picomol detection levels of target oligonucleotides. Furthermore, the QD-FRET bioassays provided for mismatch discrimination, and multiple cycles of regeneration were also demonstrated.
| Identifer | oai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/32487 |
| Date | 23 July 2012 |
| Creators | Petryayeva, Eleonora |
| Contributors | Krull, Ulrich Jorg |
| Source Sets | University of Toronto |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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