As demand for food increases, rapid testing methods are becoming increasingly important. In the past few years, yogurt has become popular. Yeast species are the most common spoilage organism, costing consumers and food companies money. A novel lateral flow assay has been developed to detect yeast oligonucleotide sequences. Gold nanoparticles were used as the standard reporter and fluorescent nanoparticles were developed as the novel reporter. The fluorescent nanoparticles were ruthenium-doped silica nanoparticles synthesized using the modified Stöber method. Visual analysis of assays using standard reporters showed the limit of detection to be 10 femtomoles of target sequence. Analysis of the fluorescent nanoparticles using a plate reader showed the limit of detection to be 0.027 femtomoles. The fluorescent reporter’s limit of detection is 1000 fold lower due to a sophisticated, more sensitive analysis method. Gold nanoparticles are appropriate for presence or absence testing, but fluorescent nanoparticles are best for obtaining quantitative data with low detection limits. Pathogens have been used as biological warfare for centuries. A brief review of common biowarfare agents is included. Yersinia pestis, the causative agent of the Plague, and Bacillus anthracis, the causative agent of Anthrax, are the focus. Additional work using gold nanoparticles as reporter in a sandwich assay is also included. The novel dye covered reporter was compared to the control, which was a single dye molecule linked to the reporter sequence. Repeated testing showed the novel reporter had a lower limit of detection and higher sensitivity due to increased ability to bind target.

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Due to enhanced properties such as large surface area-to-volume ratio, metal nanoparticles are often employed as catalysts for various applications. However, most studies involving nanoparticle catalysts have been conducted on collections of particles rather than single nanoparticles. Results obtained for ensemble systems can be difficult to interpret due to variations in particle loading and interparticle distance, which are often challenging to control and characterize. In this study, two immobilization strategies for incorporating gold nanoparticles (AuNPs) on carbon fiber ultramicroelectrodes (UMEs) were compared with the goal of extending these techniques to nanoelectrodes for studies of single AuNPs. Both layer-by-layer deposition of AuNPs on natural carbon fiber UMEs and direct reduction of AuNPs on nitrided carbon fiber UMEs were explored. Although both methods proved feasible, the direct reduction method seemed to be more effective and should better enable direct comparisons of bare and capped AuNPs.

The study of ferritin includes a rich history of discoveries and scientific progress. Initially, the composition of ferritin was determined. Soon, it was shown that ferritin is a spherical, hollow protein. Eventually, over several decades of research, the structure and some function of this interesting protein was elucidated. However, the ferritin field was not completely satisfied. Today, for example, researchers are interested in refining the details of ferritin function, in discovering the role of ferritin in a variety of diseases, and in using ferritin for materials chemistry applications. The work presented in this dissertation highlights the progress that we have made in each of these three areas: 1) Mechanistic studies: The buffer used during horse spleen ferritin iron loading significantly influences the mineralization process and the quantity of iron deposited in ferritin. The ferrihydrite core of ferritin is crystalline and ordered when iron is loaded into ferritin in the presence of imidazole buffer. On the other hand, when iron is loaded into ferritin in the presence of MOPS buffer, the ferrihydrite core is less crystalline and less ordered, and a smaller amount of total iron is loaded in ferritin. We also show that iron can be released from the ferritin core in a non-reductive manner. The rate of Fe3+ release from horse spleen ferritin was measured using the Fe3+-specific chelator desferoxamine. We show that iron release occurs by three kinetic events. 2) Disease studies: In order to better understand iron disruption during disease states, we performed in vitro assays that mimicked chronic kidney disease. We tested the hypothesis that elevated levels of serum phosphate interrupted normal iron binding by transferrin and ferritin. Results show that phosphate competes for iron, forming an iron(III)-phosphate complex that is inaccessible to either transferrin or ferritin. Ferritin samples separated from the iron(III)-phosphate complex shows that as the phosphate concentration increases, iron loading into ferritin decreases. 3) Materials chemistry studies: Anion sequestration during ferritin core reduction was studied. When the core of horse spleen ferritin is fully reduced using formamidine sulfinic acid, a variety of anions, including halides and oxoanions, cross the protein shell and enter the ferritin interior. Efforts have been made to use ferritin to control the concentration of anions for reactions. In addition, the native ferrihydrite mineral core of ferritin is a semi-conductor capable of catalyzing oxidation/reduction reactions. Light can photo-reduce AuCl4- to form gold nanoparticles (AuNPs) with ferritin as a photocatalyst. The mechanism of AuNP formation using ferritin as a photocatalyst was examined. From this work, we propose that the ferrihydrite core of ferritin photo-reduces; the mineral core dissolves into a soluble iron(II) mineral. The iron(II) then re-oxidizes, and a new mineral forms that appears to be the new photocatalyst, as the lag phase is significantly decreased with this new mineral form of ferritin.

Surface plasmon resonances of metal nanostructures have been studied intensely in recent years. The strong plasmon-mediated electric field enhancement and field confinement well beyond the diffraction limit has been demonstrated to improve the performance of optical devices including ultrasensitive sensors, light emitters, and optical absorbers. A plasmon resonance mode of particular recent interest is the gap plasmon resonance that occurs on closely spaced metallic structures. In contrast to plasmon resonances supported by isolated metal nanostructures, coupled nanostructures provide additional spectral and spatial control over the plasmon resonance response. For example, the resonance frequencies of metal nanoparticle dimers depend strongly on the gap size between the nanoparticles. Gap plasmons can produce local electric field enhancement factors that are several orders of magnitude stronger and more confined than surface plasmon resonances of isolated plasmonic nanospheres. The reliance of gap plasmons on few-nanometer separation between nanostructures makes it difficult to prepare gap-plasmon supporting structures with predictable resonance frequency and field enhancement. A structure that avoids this challenge is the film-coupled nanoparticle (NP). Similar to nanoparticle dimers, a nanoparticle on a supporting metallic film (or NP-on-a-mirror) can offer a strong coupling between the particle and its local environment, in this case the supporting film instead of adjacent nanoparticles, enabling strongly confined gap-plasmon modes. The NP-on-a-mirror geometry has been shown to produce reproducible gap plasmon resonances in a chemically and thermally robust, easy to fabricate structure. In this Thesis, we first present a scheme for controlling the gap plasmon resonance frequency of single gold nanoparticles using aluminum oxide coated metal films. We demonstrate experimentally and numerically that the gap-plasmon resonance of single gold nanoparticles can be tuned throughout the visible range by controlling the aluminum oxide thickness via anodization. In a separate study of Au NP on Al2O3 coated gold films it is shown that the oxide coating improves the stability of the structure under intense laser irradiation. An combined experimental and numerical analysis of the spectral response of Au NP on rough Au films shows that a film roughness of a few nanometer can affect the gap plasmon resonance in the absence of an oxide spacer layer. A photoluminescence study of single gold nanoparticles on an Al2O3 coated gold film shows that the gap-plasmon resonance of this type of plasmonic structure can increase gold photoluminescence by more than four orders of magnitude. Related numerical simulations reveal that the local photoluminescence enhancement of a gold nanoparticle on an Al2O3 coated gold film can be as high as one million near the particle-film junction. Finally, a new plasmonic sensing element was proposed based on our findings in the previous chapters. This proposed hole-in-one structure offers several attractive features including an easily optically accessible gap plasmon mode, while maintaining a relatively simple fabrication method. Taken together, the research presented in this Thesis demonstrates how the resonance frequency, field enhancement, mode polarization, structural stability, and structure reliability can be controlled at the nanoscale. The knowledge gained in the course of this work could lead to further development of nanophotonic devices that utilize extremely confined optical fields and precisely controlled resonance frequencies.

Localized surface plasmon resonance (LSPR) is induced in metal nanoparticles by resonance between incident photons and conduction electrons in nanoparticles. For noble metal nanoparticles, LSPR can lead to strong absorbance of ultraviolet-violet light. Although it is well known that LSPR depends on the size and shape of nanoparticles, the inter-particle spacing, the dielectric properties of metal and the surrounding medium, the temperature dependence of LSPR is not well understood. By thermally annealing gold nanoparticle arrays formed by nanosphere lithography, a shift of LSPR peak upon heating has been shown. The thermal characteristics of the plasmonic nanoparticles have been further used to detect chemicals such as explosive and mercury vapors, which allow direct visual observation of the presence of mercury vapor, as well as thermal desorption measurements

This dissertation demonstrated that the manipulation of substances at the molecular or nanometer level can lead to the discovery and development of new materials with interesting properties and important applications. Chapter 1 describes the development of a nanoscale molecular thin film material for corrosion protection. By using a self-assembled monolayer film with a thickness of only about 1 nanometer as a linkage, a covalent bonding was achieved between a polyurethane top coating and an aluminum alloy substrate. This covalent bonding between polymer top coating and the aluminum alloy substrate significantly improved the corrosion resistance of the substrate. Chapter 2 and Chapter 3 describe the development of a gold nanoparticle-polymer composite material in different forms with a number of applications. Gold nanoparticles are among one of the most extensively studied nanomaterials. When the size of gold is shrunk to the nanometer scale, many interesting and new physical properties start to appear from gold nanoparticles. The optical properties of gold nanoparticles, particularly the surface plasmon resonance absorption, have been investigated in this dissertation for the development of multifunctional nanocomposite materials. Chapter 2 presents the preparation of a gold nanoparticle/poly(methyl methacrylate) (PMMA) nanocomposite film and the application of such films for microstructure fabrication using a direct laser writing technique. Gold nanoparticles are excellent photon-thermal energy converters due to their large absorption cross section at the surface plasmon resonance region. Upon laser irradiation of the nanocomposite film, the thermal energy converted from the absorbed photon energy by gold nanopaticles induced a complete decomposition of PMMA, leading to the formation of various microstructures on the nanocomposite films. Chapter 3 reports the further development of a nanoparticle/polymer composite nanofiber material fabricated through an electrospinning process. The matrix of the nanofiber is made of two polyelectrolytes, poly(acrylic acid) (PAA) and poly(allylamine hydrochloride) (PAH). Three methods were developed to incorporate gold nanoparticles into the polymer matrix. The composite nanofiber materials developed in this study demonstrate multifunctional properties, including good electrical conductivity, photothermal response, and surface-enhanced IR absorption. This material may be used for many important applications including catalysis, chemical and biological sensors, and scaffold materials for tissue engineering. In Chapter 4, another most important nanomaterial, carbon naotubes (CNTs), were introduced as fillers to prepare polymer nanocomposites. A dispersion method for multi-walled carbon nanotubes (MWCNTs) using a conjugated conducting polymer, poly(3-hexylthiophene) (P3HT) as the third component and trifluoroacetic acid (TFA) as a co-solvent was developed. Due to the excellent dispersion of carbon nanotubes in PMMA and enhanced conductivity of the nanocomposites by the conjugated conducting polymers, the prepared composite materials has an extremely low percolation threshold of less than 0.006 wt% of MWCNT content. The potential use of MWCNT/conducting polymer composites for energy storage applications such as suppercapacitors was further investigated by Cyclic Voltammetry (CV), Electrochemical Impedance Spectroscopy (EIS) and charging-discharging cycles. Compared to pure carbon nanotubes, the nanocomposite materials have significantly improved properties and are promising for supercapacitor applications.

Lysosomen spielen entscheidende Rolle bei der zellulären Homöostase. Die Überwachung von Lysosomen, die Lipide ansammeln, ist eine erhebliche Herausforderung. Diese Arbeit konzentriert sich auf die Entwicklung der oberflächenverstärkten Raman-Streuung (SERS) als Methode zur Überwachung intakter Lysosomen, insbesondere hinsichtlich des Einflusses von Arzneimitteln, die den Lipidstoffwechsel stören. Um das Potenzial von SERS zur Untersuchung von Lysosomen in lebenden Zellen zu bewerten, wurden die Wechselwirkungen zwischen trizyklischen Antidepressiva und saurer Sphingomyelinase untersucht. Zunächst wurden Modellsysteme untersucht. Die Wechselwirkungen zwischen den Antidepressiva und Goldnanopartikeln wurden durch SERS charakterisiert. Die Daten zeigten, dass Moleküle mit den Nanopartikeln interagieren. Als Modellsystem der lipidreichen Umgebung wurden Komposite aus Liposomen und Goldnanopartikeln von SERS und Cryo-EM untersucht. Die SERS-Spektren sind charakteristisch für die Lipidzusammensetzung der Vesikel. Die Wechselwirkungen zwischen den Antidepressiva und den Lysosomen wurden in der Fibroblastenzelllinie 3T3 durch SERS und komplementäre Methoden untersucht. In Übereinstimmung mit den SERS-Spektren von Modellsystemen zeigen die SERS-Spektren lebender Zellen Signaturen sowohl der Antidepressiva als auch der Lipide. Um die Unterschiede in den Lysosomen zwischen behandelten und nicht behandelten Zellen aufzudecken, wurde ein zufälliger Waldansatz verwendet. Darüber hinaus wurde SERS verwendet, um die Lipidverteilung in Leishmania-infizierten Makrophagen zu untersuchen, von denen bekannt ist, dass sie Lipide akkumulieren. Die Ergebnisse zeigen, dass SERS verwendet werden kann, um die Lipidzusammensetzung in lebenden Zellen verschiedener Zelltypen zu untersuchen. Als neue methodische Entwicklung zeigt die Random-Forest-Analyse von SERS-Daten, dass Ansätze des maschinellen Lernens für ein besseres Verständnis von Daten aus biologischen Systemen nützlich sein können. / Lysosomes play a crucial role in cellular homeostasis. Monitoring lysosomes that accumulate lipids represents a considerable challenge. This thesis focuses on the development of surface-enhanced Raman scattering (SERS) as a method to monitor intact lysosomes, in particular regarding the influence of drugs that interfere with lipid metabolism. To evaluate the potential of SERS for studying lysosomes in live cells, the interactions between tricyclic antidepressants and acid sphingomyelinase were studied. First, model systems were investigated. The interactions between the antidepressants and gold nanoparticles were characterized by SERS. The data showed that molecules interact with the nanoparticles. As a model system of the lipid-rich environment, composites of liposome and gold nanoparticles were studied by SERS and cryo-EM. The SERS spectra are characteristic of the vesicles’ lipid composition. The interactions between the antidepressants and the lysosomes were studied in the fibroblast cell line 3T3 by SERS and complementary methods. In agreement with the SERS spectra of model systems, the SERS spectra of live cells show signatures of both, the antidepressants and the lipids. To reveal the differences in the lysosomes between treated and non-treated cells, a random forest approach was used. Moreover, SERS was used to study the lipid distribution in Leishmania-infected macrophages known to accumulate lipids. The results show that SERS can be used to investigate lipid composition in live cells of different cell types. As a new methodological development, the random forest analysis of SERS data shows that machine learning approaches can be useful for a better understanding of data from biological systems.