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Immobilizations for an aluminum nitride (AlN) based biosensor and real-time polymerase chain reaction (RT-PCR) device

Posted on:2008-04-13Degree:Ph.DType:Dissertation
University:Wayne State UniversityCandidate:Hughes, ChantelleFull Text:PDF
GTID:1444390005463195Subject:Engineering
Abstract/Summary:
Aluminum nitride (AlN) has many characteristics that make it a desirable material for AW sensor development, such as its piezoelectric coupling properties and linear temperature coefficient. A novel AlN-based SAW/STW sensor has been developed, thereby, it can detect in gaseous and liquid mediums, respectively. This ability to operate in a liquid medium offers the potential for use as a biological detection system. An immobilization region (bound chemicals and/or proteins to the sensor surface) is necessary to transform it into a functional biosensor. The first part of this study examines the utilization of (3-aminopropyl)trimethoxysilane (APTMS) monolayers on AlN wafers, for the immobilization of antibodies and protein detection. The ability of this monolayer to act as a binding layer for antibodies was evaluated by XPS, enzyme-linked immunosorbent assay (ELISA), AFM, and fluorescence microscopy.;Working towards the goal of developing a portable, PCR-based device for the quantification of cellular damage due to radiation exposure in an individual, in human lymphocytes, 20+ dose-dependent genes and 6 dose-independent genes have been identified and prototype system for their analysis has been developed. The prototype device is capable of performing all genetic analysis with only a drop of blood. It consists of a few main components: (1) cell lyses region (genetic materials released from lymphocytes), (2) immobilization region for mRNA isolation, and (3) a real-time PCR portion, for gene amplification and analysis. The second part of this study focuses on the development of the immobilization region, for the Pyrex-based device. The device's design prompted silanization by a liquid method, with 10-(carbomethoxy)decyl-dimethylchlorosilane. Oligonucleotides were covalently immobilized to the silane, so mRNA could be captured and isolated in the device. The silane is commercially available and to our knowledge has not been utilized for surface-nucleic acid attachment. Analysis of the binding process for the mRNA isolation and preliminary results from the testing of the prototype device will be presented.
Keywords/Search Tags:Device, Aln, Sensor, Immobilization
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