This master thesis deals with conventional methods of bacterial cell analysis, polyhydroxyalkanoates, Raman spectroscopy and electron microscopy in the theoretical part. The production of polyhydroxybutyrate by selected thermophilic bacteria and their analysis by gas chromatography, cryogenic scanning electron microscopy and Raman spectroscopy is described in the experimental part. The chosen sample was analyzed by a transmission electron microscope. Comparing the results from previous mentioned methods it was found that the bacteria Schlegelella thermodepolymerans accumulated the highest amount of PHB. The lowest amount of PHB was obtained by bacteria Rubrobacter xylanophilus. The assumption that the PHB granules formed so-called needle-like plastic deformations during freeze-fracturing was affirmed by cryo-SEM photos analysis. Moreover, it was found that the bacterial cell characterization deduced from microscopic observation of samples corresponded to the description in the literature. TEM provided better resolution photos and in consequence the cells and PHB are more visible. The thesis is also focused on chemical fingerprint analysis of cells by Raman spectroscopy. Several biomolecules were identified by measured Raman spectra for the particular samples.

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A biosensor is an analytical device integrating a biological element and a physicochemical transducer that convert a biological response into a measurable signal. The advantages of biosensors include low cost, small size, quick, sensitivity and selectivity greater than the conventional instruments. Biosensors have a wide range of applications ranging from clinical diagnostics through to environmental monitoring, agriculture industry, et al. The different types of biosensors are classified based on the sensor device as well as the biological material. Biosensors can be broadly classified into (piezoelectric, etc.), electrochemical biosensors (potentiometric, amperometric, etc.), and optical types of biosensors (fiber optics, etc.). Here, we introduce a novel microfluidics-integrated biosensor platform system that can be flexibly adapted to form individual biosensors for different applications. In this dissertation, we present five examples of different emerging areas with this biosensor system including anti-cancer drug screening, glucose monitoring, heavy metal elements measurement, obesity healthcare, and waterborne pathogen DNA detection. These micro-biosensors have great potential to be further developed to emerging portable sensing devices especially for the uses in the developing and undeveloped world. At the last chapter, Raman spectroscopy applied to assess gestational status and the potential for pregnancy complications is presented and discussed. This technique could significantly benefit animal reproduction.

Les composants HEMTs (High Electron Mobility Transistors) à base d’AlGaN/GaN sont à ce jour les candidats les plus prometteurs pour des applications hyperfréquences de puissance, dû essentiellement à leur forte densité de porteurs et des mobilités électroniques élevées. Cependant, la température générée en condition réelle est un paramètre capital à mesurer, afin d’estimer précisément la fiabilité des composants et leur durée de vie. Pour ces raisons, nous avons comparé les méthodes de caractérisation thermique par thermoréflectance et par spectroscopie Raman car elles sont non destructives et avec une résolution spatiale submicronique. Ces techniques ont déjà prouvé leur faisabilité pour la caractérisation thermique des transistors, en modes de fonctionnement continu et pulsé. Nous comparons dans cette étude leurs adaptabilité et performance dans le cadre de la réalisation d’un banc d’essai thermique dédié. Ces méthodes sont reconnues pour ne caractériser que certaines catégories de matériaux : les métaux pour la thermoréflectance et les semiconducteurs pour la spectroscopie Raman, ce qui nous a conduit à l’éventualité de les combiner. Nous avons confronté des résultats obtenus par thermoréflectance à partir des équipements de deux fabricants commercialisant cette méthode, nous permettant ainsi de mettre en évidence des résultats originaux sur des aspects et inconvénients qui ne sont pas relayés dans la littérature. Avec la spectroscopie Raman, nous avons identifié les paramètres de métrologie qui permettent de réaliser un protocole de mesure thermique le plus répétable possible, et nous présentons également une technique innovante pour sonder les matériaux en surface, à l'aide du même équipement, et notamment les métaux. / At the moment, AlGaN/GaN HEMTs (High Electron Mobility Transistors) are the most promising for high-power hyperfrequency applications, essentially due to their large carrier density and a high electronic mobility. However, the temperature generating during operational conditions is a crucial parameter to measure, in order to estimate the reliability and durability of components. For these reasons, we compared thermoreflectance and Raman spectroscopy, that are non-destructive and possessing a submicronic spatial resolution. These techniques have already proven their feasibility as thermal characterization methods in both continuous wave and pulsed operational modes. We compare here their adaptability and performance to the conception of a thermal test bench. These methods are known for characterizing specific types of material: metals for thermoreflectance and semiconductors for Raman spectroscopy, leading us to the eventuality to combine them. We compared several results measured by thermoreflectance method with equipment from two different manufacturers that commercialize this technology, so we could highlight some aspects and drawbacks that are note relayed in the literature. With Raman spectroscopy, we identified metrology parameters allowing to realize a thermal measurement setup as reproducible as possible, and we also present an innovative method to probe surface material, especially metals.

Graphene research is nowadays one of the worldwide most prominent fields of interest in material science due to many extraordinary properties of graphene and related materials. However, the different techniques of synthesis and subsequent handling and/or treatment have a substantial impact on the properties of the graphene and thus a lot of efforts have been focused on developing of the advanced methods for graphene preparation and characterization. Graphene can be easily produced by oxidation and consequent exfoliation of the bulk graphite; however, resulting graphene oxide needs to be reduced back to graphene-like structure due to partial restoration of sp2 network. Herein, a detailed study of the structural evolution of the graphene oxide during electrochemical treatment has been performed using X-ray photoelectron, Raman and infrared spectroscopies and the results were compared with non-oxidized graphene nano-platelets. Additionally, graphene oxide in composite with LiFePO4 olivine material, which is electrochemically almost inactive in a freshly made state, has been tested by repeated electrochemical cycling. Using various electrochemical methods, the progressive electrochemical activity enhancement has been observed and spontaneous graphene reduction was identified as responsible for this...

There has been a notable rise in the alternative protein market in the recent years which promotes an interest in the research of both animal and plant proteins to establish better structure-function relationships. Over the years many analytical tools have been used to study proteins and compare them, however Raman Spectroscopy and Surface Enhanced Raman Spectroscopy (SERS) have not been as much used for this application. SERS consolidates Raman Spectroscopy that primarily measures molecular vibrations and nanostructures that enhance the weak Raman signals. The objective of this study is to explore the capability of the Raman instrumentation in combination with different substrates for spectroscopic analysis of 3 animal proteins viz. whey, k-casein and albumin from chicken egg white and 4 plant proteins namely mung bean, soy, pea and faba bean. Herein, we firstly established a method that could be applied to all proteins to detect characteristic peaks that are related to their structure. Of all the methods, SERS with silver dendrites was the most promising method that detected protein characteristic peaks, particularly the shifts around 700-900 cm-1 attributed to the CN stretch and tryptophan groups. Although different proteins exhibit similar spectral characteristics, they were discriminated using principal component analysis. Then we explored the optimal method to study the effect of different environmental conditions including pH and salt concentration on the protein spectroscopic analysis. The limitations of the substrates were better understood during this process as Ag dendrites failed to provide a spectrum in the high pH range but was compatible with different salt concentrations. The peaks in the Amide-I region were vi used as a marker to study the effect of change in pH and salt. Most proteins showing a shift in the band suggesting a transition from α-sheet to a random coil conformation. The acquired spectra and subsequent PCA results depicted that pea protein was the most susceptible to change in pH amongst other proteins whereas faba bean was susceptible to a change in salt concentration. Finally, these learnings were applied to analyze a real-world food product to compare its spectroscopic characteristic with the standards we have. In conclusion, we demonstrated that Raman Spectroscopy and SERS was able to provide distinct spectroscopic characteristics of plant and animal proteins that may be used to facilitate the quality control or product development of novel plant-based food products. Future work will investigate the relationship between the spectroscopic characteristics and the structural function of proteins.

At Nouryon Stockvik the Expancel production site is located. Expandable microspheres areused in for many types of applications and is a technically challenging product. At ExpancelStockvik they are constantly striving to improve product properties in line with customerexpectations. To be able to do this, it is important to understand the properties of themicrospheres such as crystallinity and crystallite size.Films was prepared from microspheres dissolved in DMA and analyzed with High-resolutionSEM, Powder X-Ray Diffraction and Raman Spectroscopy. The aim was to develop a methodto investigate and determine crystalline ratios and crystallite size within the microsphere filmsand the microspheres before film preparation. The eventual correlation between morphologyand crystallinity was also studied. An attempt of finding an amorphous reference sample wasalso performed, this was done by grinding microspheres in liquid nitrogen, unfortunately, nosuccess was reached. Gauss-fitting was therefore performed to be able to find the amorphousregions of the XRD Diffractogram for the calculations of crystallinity and crystallite size. TheGauss-fitting was successfully performed with good R-square values.During the Raman analysis some fluorescence problems occurred, this problem will probablybe solved if a laser source with higher excitation frequency is used in future analysis. Evenwith fluorescence problems, Raman analysis could successfully be performed and giveinformation of the composition. The crystallite size was in general larger for the microspheresbefore they were prepared from dissolving them to make films. Generally, it seems as there isa correlation between the morphology, crystallinity, and crystallite size.

The novel method herein proposed, aims to study Deoxyribonucleic acid (DNA) and Rad51 repair protein in its resting state after their interaction by using a combination of biological preparation and physical measures. Rad51 is a highly conserved protein; it is involved in eukaryotes genome stability and can interact with single strand (ss) and double strands (ds) DNA. In our work, a droplet of the solution containing the dsDNA/Rad51 complexes was deposited on micro-fabricated super-hydrophobic substrates (SHS) to obtain self-organized and suspended fibers. The silicon-based SHS were designed to incorporate a regular circular array of pillars and to maintain a high contact angle with the drop. The samples were let dehydrate at controlled temperature and humidity conditions. At the end of the buffer evaporation process, non-suspended material and salt excess are concentrated on the top of a few micro-pillars in a limited area (drop residual) of the device while ordered and self-assembled DNA/Rad51 fibers are suspended between micro-pillars. To find the ideal conditions to obtain and suspend the nucleic acid/protein complexes, several parameters were investigated: saline buffer, DNA and protein concentrations were widely titrated and showed a significant effect on the biomolecule suspension on SHS. The samples were then preliminarily checked by microscopy techniques and then described by the Raman spectra acquired. Several techniques were used: optical microscopy, Energy Dispersive X-Ray Spectroscopy (EDAX), Scanning Electron Microscopy (SEM) and Raman Spectroscopy. Protein expressions, DNA suspension, micro-fabrication and characterization were all performed in KAUST Core Labs and Structural Molecular Imaging Light Enhanced Spectroscopies (SMILEs) Lab. The novel approach presented in this work is highly multidisciplinary and comprises physical measurements (Raman spectroscopy and EM imaging), chemistry and biology. In future the method can be used further expanded supporting the data with HRTEM direct imaging to elucidate the nucleic acids/proteins behavior in the multiple phases of the genome repair processes. Also, it and can serve as a fingerprint of the biological molecules involved in biological interactions, their localization and structural characterization, providing a new tool for structural analysis, screening and diagnostics.

The rational combination of dielectric and metallic nano particles brings novel optical properties to conventional subwavelength structures. This thesis introduces the optoplasmonic geometries demonstrating versatile ability in both far and near field modification within nano scale. Template-assisted self-assembly approaches are applied creating nano entities with titanium dioxide and gold nano spheres. A top-bottom mono hybrid unit and interdigitated array are developed. With the examination of the elastic and inelastic response of these hybrid materials, physical models are simulated to depict the scenario of varied geometry and combination of nano particles. In contrast to solely metal or dielectric particle arrays, this type of artificial material not only enhances the near electric field intensity within the metal nano cluster hot spots, but also expands the overall volume of enhanced electric field. Further study reveals that the additional enhancement and redistribution of near field are derived from the coupling between the nano gold cluster plasmon resonance and the in-plane diffractive mode of the dielectric array. The redirected emission profile of the fluorescent dyes within the hybrid array is explored.

Indiana University-Purdue University Indianapolis (IUPUI) / Background: Stimulated Raman spectroscopy (SRS) is a nondestructive tool for biochemical characterization of tissues. The aims were: 1) To evaluate the ability of SRS and Spontaneous Raman spectroscopy (SpRS) to differentiate among sound, demineralized and remineralized bovine enamel by phosphate and carbonate ratio (P/C-Ratio); and 2) To determine the correlation between the outcomes of transverse microradiography (TMR: Integrated mineral loss (ΔZ) and lesion depth) and P/C-Ratio. Material and Methods: Thirty, 5×5×2-mm ground and polished bovine enamel blocks were prepared. The surface was divided into 3 equal areas. Each area was chemically demineralized (demin) by Carbopol demineralized solution for 0 (Sound-Demin), 24 (24h-Demin), and 48h (48h-Demin), respectively. Then, specimens were sectioned for TMR analysis, and the remaining one part of each specimen was remineralized (remin) for 15days using a pH-cycling model (Sound-Remin, 24hD-Remin=24h-Demin and remineralization, 48hD-Remin = 48h-Demin and remineralization). Demin and remin groups were scanned to obtain P/C-Ratio by SpRS and SRS. SRS was further scanned from 0 (surface) up to 100 µm into the dentine at 10-µm intervals. Remineralized specimens were sectioned for TMR analysis. Wilcoxon signed-rank tests were used to compare between TMR and SpRS/SRS. Spearman correlation coefficients were used to correlate among TMR, SpRS, and SRS. A 5-percent significance level was used for each test. Results: As demin time increased, both ΔZ and lesion depth were increased. After remineralization, both values were decreased. There were significant differences between demine and remin groups and between demin times. For SpRS, Sound-Demin had significantly larger P/C-Ratio than 24h-Demin and 48h-Demin (p ≤ 0.001). The 24h-Demin had significantly larger values than 48h-Demin (p = 0.048). Sound-Remin had larger P/C-Ratio than 24hD-Remin (p = 0.316) and 48hD-Remin (p = 0.015). 24hD-Remin was larger than 48hD-Remin (p = 0.269). 24hD-Remin had significantly larger P/C-Ratio than 24h-Demin (p ≤ 0.001). 48hD-Remin had significantly larger P/C-Ratio than 48h-Demin (p ≤ 0.001). For SRS, at surface (0 µm), for demin group, Sound-Demin had significantly larger P/C-Ratio than 24h-Demin (p = 0.020) and 48h-Demin (p = 0.032). 24h-Demin had larger value than 48h-Demin; but no significant difference (p = 0.117). Among remin groups, Sound-Remin was not statistical significance different for 24hD-Remin (p = 0.172) and 48hD-Remin (p = 0.134). However, 24hD-Remin was smaller; but not statistical significance different from 48hD-Remin (p = 0.688). At deeper levels (10 µm to 100µm), it was found that 1) After demineralization, Sound-Demin had significantly larger P/C-Ratio than 24h-Demin and 48h-Demin at 0 µm to 20 µm, and 80 µm to 100µm; Sound-Demin had significantly larger P/C-Ratio than 48h-Demin; and no statistical significance differences were found among Sound-Demin and 24h-Demin, 24h-Demin and 48h-Demin. 2) After remineralization, no statistical significance differences were found among Sound-Remin, 24hD-Remin, and 48hD-Remin. 3) Sound-Demin had significantly larger P/C-Ratio than Sound-Remin at 0 µm ,10 µm, 20 µm; and no statistical significance differences were found at levels deeper than 30 µm. 4) No statistical significance differences were found between 24h-Demin and 24hD-Remin from 0 µm to 70µm; and 24hD-Remin had significantly larger P/C-Ratio than 24h-Demin from 80 µm to 100 µm. 5) No statistical significance differences were found between 48h-Demin and 48hD-Remin. For correlation, moderate correlation was found between SpRS demineralized/remineralized groups and ΔZ, and between SpRS demineralized groups and lesion depth. Conclusion: SpRS and SRS have the potential to quantify demineralization through calculation of the phosphate and carbonate ratio. In addition, SpRS can detect the change of remineralization. A nondestructive caries detection approach using SpRS and SRS would be beneficial in clinical practice.