| In the first part of the thesis our work on a surface plasmon resonance (SPR) biosensor will be presented. It will begin with understanding the working principle of SPR sensing technology and the basic concept of SPR biosensing. In SPR technology, there are different coupling schemes to excite surface plasmons such as prism coupler, grating coupler and waveguide coupler. Our setup will be based on the attenuated total reflection (ATR) prism coupling configuration. A gold sensing film is attached to one face of the prism. The samples are flowing over the gold surface and the light source is directed to the prism side. The reflected beam containing SPR information is collected and analyzed. SPR biosensors have become powerful tools in biological and chemical sensing application because of their capability of real-time monitoring and label-free sensing. Quantitative measurements such as the binding kinetics and the binding affinity between two biomolecules can be readily calculated from the SPR sensorgram. In our design, SPR phase will be monitored using photoelastic modulation (PEM) technique. The PEM is used to produce a modulation signal so that the phase quantity can be extracted by measuring the relative amplitudes of the harmonic signals. Since this system contains no moving component and only single beam and single detector are used, precise component alignment, which may be troublesome in making the setup compact and robust, can be eliminated. In order to demonstrate the operation of the proposed approach, two experiments were performed. The first one was to measure the refractive index change caused by varying the concentration of glycerin-water mixtures. The second one was to monitor the binding reactions between biotin and streptavidin--BSA complex at the sensor surface. Recently, the use of metallic nanoparticle on SPR platform has received great attention due to the capability of sensitivity enhancement. Although the mechanism of the enhancement is still not fully understand, three possible factors are concluded after systematic researches: (i) an increase of the absolute mass in each binding event, (ii) an increase in the bulk refractive index of the analyte, and (iii) coupling between the localized surface plasmon resonance (LSPR) of metallic nanoparticles and surface plasmon resonance (SPR) of the sensing film. Indeed, the role of plasmonic coupling in sensitivity enhancement is still an open question. In order to obtain a better understanding of this phenomenon, at the end of part I, extended studies were performed to investigate how the LSPR properties of metallic nanoparticle labels correlate with the enhancement factor. For this purpose, gold nanorods (Au-NRs) were chosen as the amplification labels because of the easy tunability of LSPR peak of Au-NR. After reading the "Result and Discussion" section, the readers will have better understanding of "plasmonic coupling" between the sensing film and the metallic labels with suitable operating laser source.;In the second part of the thesis, the bioimaging part, the application of nanostructure materials in live cancer cell imaging and small animal imaging were demonstrated. There are different types of imaging technique available in laboratories and clinics: optical imaging, computed tomography (CT), magnetic resonance imaging (MRI), positron emission tomography (PET), thermography and ultrasound imaging. Although such imaging techniques have been well developed and used over a decade, improving the sensitivity, enhancing the contrast, decreasing the acquisition time and reducing the toxicity of the contrast agent are highly desirable. For optical imaging, the scientists discovered that the use of near infrared fluorescence materials can assist the surgeon to locate the tumor, the nerve and the lymph node more accurately. For CT scan, the use of Au-NR as the contrast agent can improve the sensitivity. Iron oxide nanoparticle or gadolinium ion containing nanoparticle can greatly enhance the contrast of MRI. On the one hand, concrete effort has been concentrated on exploring the feasibilities of nanomaterials. However, on the contrary, the researchers also revealed the cytotoxicity of nanoparticles and the potential hazard for long term circulation in vivo. They argued that the long-term contact of the nanoparticles with biological fluids can result partial desorption of the hydrophilic moieties, thus exposing the bare surface to the biological system, with high chances of releasing toxic ions to the surrounding. This dissertation will focus on two nanomaterials, Au-NR and QD, using as nanoprobes for live pancreatic cancer cells imaging and small animal imaging. Different surface modification strategies and the biocompatibility will be discussed. The toxicities of the nanomaterials will also be evaluated by appropriate bio-assay. (Abstract shortened by UMI.)... |
