Immunofluorescence assays are capable of both detecting the amount of a protein and the location of the protein within a cell or tissue section. Unfortunately, the traditional technique is not capable of detecting concentrations on the nanoscale. Also, the technique suffers from non-specific attachment, which can cause false-positives, as well as photobleaching when detecting lower concentrations is attempted. There is also a time constraint problem since the technique can take from many hours to a few days in some cases.
In this work, metal-enhanced fluorescence (MEF) is used to lower the detection limit and reduce photobleaching. Unfortunately, MEF also increases the intensity of non-specifically bound proteins (NSBPs). Therefore, a surface acoustic wave (SAW) device is used to remove the more weakly bound NSBPs. Previously, this has been shown on lithium niobate, but it is used with a quartz substrate in this work. The SAW device is also used to cause micro-mixing which speeds the process up significantly.
In this research, it was found that silver nanocubes can lower the detection limit down to below 1 ng/mL. Quartz SAW devices are shown to remove NSBPs at a power of 10 mW applied for five minutes. Micro-mixing is shown to be improved by a factor of six at 10 mW for 10 minutes by saturating the antibody used in this research, which takes 1 hour without micro-mixing. Finally, all three components are combined. In this work, the whole device is used to detect 50 ng/mL. After micro-mixing, the intensity is the same as with MEF, and, after removal, it has been lowered by 7 a.u.
| Identifer | oai:union.ndltd.org:USF/oai:scholarcommons.usf.edu:etd-6277 |
| Date | 17 March 2014 |
| Creators | Morrill, Samuel |
| Publisher | Scholar Commons |
| Source Sets | University of South Flordia |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Graduate Theses and Dissertations |
| Rights | default |
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