Particle analysis using multiwavelength UV-visible spectroscopy provides the potential for extracting quantitative red blood cell information, such as hemoglobin concentration, cell size, and cell count. However, if there is a significant presence of hypochromism as a result of the concentrated hemoglobin (physiological value of 33%), successful quantification of red cell values would require a correction. Hypochromism has been traditionally defined as a decrease in absorption relative to the values expected from the Beer-Lambert Law due to electronic interactions of chromophores residing in close proximity of one another. This phenomenon has been suggested to be present in macroscopic systems composed of strong chromophores such as nucleic acids, chlorophyll, and hemoglobin. The study presented in this dissertation examines the presence of hypochromism in red blood cells as a part of a larger goal to qualitatively and quantatively characterize red blood cells using multiwavelength UV-visible spectroscopy. The strategy of the study was three-fold: 1) to determine the instrumental configuration that would provide the most complete information in the acquired spectra, 2) to develop an experimental model system in which the hemoglobin content in red blood cells could be modified to various concentrations, and 3) to implement an interpretation model based on light scattering theory (which accounts for both the scattering and absorption components of the optical density spectrum) to provide quantitative information for the experimental system. By this process, hypochromicity was redefined into two categories with molecular hypochromicity representing the traditional definition and macroscopic hypochromicity being an attenuation of the absorption component due to a scattering-related effect. Successful simulations of experimental red cell spectra containing various amounts of hemoglobin were obtained using the theoretical model. Furthermore, successful quantitative interpretation of the red blood cell spectra was achieved in the context of corpuscular hemoglobin concentration, corpuscular volume, and cell count solely by accounting for the scattering and absorption effects of the particle, indicating that molecular hypochromicity was insignificant in this macroscopic system.

The determination of the atmospheric aerosol size distribution is an inverse illposed problem. The shape and the material composition of the air-carried particles are two substantial model parameters. Present evaluation algorithms only used an approximation with spherical homogeneous particles. In this paper we propose a new numerically efficient recursive algorithm for inhomogeneous multilayered coated and absorbing particles. Numerical results of real existing particles show that the influence of the two parameters on the model is very important and therefore cannot be ignored.

Analytical ultracentrifugation (AUC) has made an important contribution to polymer and particle characterization since its invention by Svedberg (Svedberg and Nichols 1923; Svedberg and Pederson 1940) in 1923. In 1926, Svedberg won the Nobel price for his scientific work on disperse systems including work with AUC. The first important discovery performed with AUC was to show the existence of macromolecules. Since that time AUC has become an important tool to study polymers in biophysics and biochemistry. AUC is an absolute technique that does not need any standard. Molar masses between 200 and 1014 g/mol and particle size between 1 and 5000 nm can be detected by AUC. Sample can be fractionated into its components due to its molar mass, particle size, structure or density without any stationary phase requirement as it is the case in chromatographic techniques. This very property of AUC earns it an important status in the analysis of polymers and particles. The distribution of molar mass, particle sizes and densities can be measured with the fractionation. Different types of experiments can give complementary physicochemical parameters. For example, sedimentation equilibrium experiments can lead to the study of pure thermodynamics. For complex mixtures, AUC is the main method that can analyze the system. Interactions between molecules can be studied at different concentrations without destroying the chemical equilibrium (Kim et al. 1977). Biologically relevant weak interactions can also be monitored (K ≈ 10-100 M-1). An analytical ultracentrifuge experiment can yield the following information: • Molecular weight of the sample • Number of the components in the sample if the sample is not a single component • Homogeneity of the sample • Molecular weight distribution if the sample is not a single component • Size and shape of macromolecules & particles • Aggregation & interaction of macromolecules • Conformational changes of macromolecules • Sedimentation coefficient and density distribution Such an extremely wide application area of AUC allows the investigation of all samples consisting of a solvent and a dispersed or dissolved substance including gels, micro gels, dispersions, emulsions and solutions. Another fact is that solvent or pH limitation does not exist for this method. A lot of new application areas are still flourishing, although the technique is 80 years old. In 1970s, 1500 AUC were operational throughout the world. At those times, due to the limitation in detection technologies, experimental results were obtained with photographic records. As time passed, faster techniques such as size exclusion chromatography (SEC), light scattering (LS) or SDS-gel electrophoresis occupied the same research fields with AUC. Due to these relatively new techniques, AUC began to loose its importance. In the 1980s, only a few AUC were in use throughout the world. In the beginning of the 1990s a modern AUC -the Optima XL-A - was released by Beckman Instruments (Giebeler 1992). The Optima XL-A was equipped with a modern computerized scanning absorption detector. The addition of Rayleigh Interference Optics is introduced which is called XL-I AUC. Furthermore, major development in computers made the analysis easier with the help of new analysis software. Today, about 400 XL-I AUC exist worldwide. It is usually applied in the industry of pharmacy, biopharmacy and polymer companies as well as in academic research fields such as biochemistry, biophysics, molecular biology and material science. About 350 core scientific publications which use analytical ultracentrifugation are published every year (source: SciFinder 2008 ) with an increasing number of references (436 reference in 2008). A tremendous progress has been made in method and analysis software after digitalization of experimental data with the release of XL-I. In comparison to the previous decade, data analysis became more efficient and reliable. Today, AUC labs can routinely use sophisticated data analysis methods for determination of sedimentation coefficient distributions (Demeler and van Holde 2004; Schuck 2000; Stafford 1992), molar mass distributions (Brookes and Demeler 2008; Brookes et al. 2006; Brown and Schuck 2006), interaction constants (Cao and Demeler 2008; Schuck 1998; Stafford and Sherwood 2004), particle size distributions with Angstrom resolution (Cölfen and Pauck 1997) and the simulations determination of size and shape distributions from sedimentation velocity experiments (Brookes and Demeler 2005; Brookes et al. 2006). These methods are also available in powerful software packages that combines various methods, such as, Ultrascan (Demeler 2005), Sedift/Sedphat (Schuck 1998; Vistica et al. 2004) and Sedanal (Stafford and Sherwood 2004). All these powerful packages are free of charge. Furthermore, Ultrascans source code is licensed under the GNU Public License (http://www.gnu.org/copyleft/gpl.html). Thus, Ultrascan can be further improved by any research group. Workshops are organized to support these software packages. Despite of the tremendous developments in data analysis, hardware for the system has not developed much. Although there are various user developed detectors in research laboratories, they are not commercially available. Since 1992, only one new optical system called “the fluorescence optics” (Schmidt and Reisner, 1992, MacGregor et al. 2004, MacGregor, 2006, Laue and Kroe, in press) has been commercialized. However, except that, there has been no commercially available improvement in the optical system. The interesting fact about the current hardware of the XL-I is that it is 20 years old, although there has been an enormous development in microelectronics, software and in optical systems in the last 20 years, which could be utilized for improved detectors. As examples of user developed detector, Bhattacharyya (Bhattacharyya 2006) described a Multiwavelength-Analytical Ultracentrifuge (MWL-AUC), a Raman detector and a small angle laser light scattering detector in his PhD thesis. MWL-AUC became operational, but a very high noise level prevented to work with real samples. Tests with the Raman detector were not successful due to the low light intensity and thus high integration time is required. The small angle laser light scattering detector could only detect latex particles but failed to detect smaller particles and molecules due to low sensitivity of the detector (a photodiode was used as detector). The primary motivation of this work is to construct a detector which can measure new physico-chemical properties with AUC with a nicely fractionated sample in the cell. The final goal is to obtain a multiwavelength detector for the AUC that measures complementary quantities. Instrument development is an option for a scientist only when there is a huge potential benefit but there is no available commercial enterprise developing appropriate equipment, or if there is not enough financial support to buy it. The first case was our motivation for developing detectors for AUC. Our aim is to use today’s technological advances in microelectronics, programming, mechanics in order to develop new detectors for AUC and improve the existing MWL detector to routine operation mode. The project has multiple aspects which can be listed as mechanical, electronical, optical, software, hardware, chemical, industrial and biological. Hence, by its nature it is a multidisciplinary project. Again by its nature it contains the structural problem of its kind; the problem of determining the exact discipline to follow at each new step. It comprises the risk of becoming lost in some direction. Having that fact in mind, we have chosen the simplest possible solution to any optical, mechanical, electronic, software or hardware problem we have encountered and we have always tried to see the overall picture. In this research, we have designed CCD-C-AUC (CCD Camera UV/Vis absorption detector for AUC) and SLS-AUC (Static Light Scattering detector for AUC) and tested them. One of the SLS-AUC designs produced successful test results, but the design could not be brought to the operational stage. However, the operational state Multiwavelength Analytical Ultracentrifuge (MWL-AUC) AUC has been developed which is an important detector in the fields of chemistry, biology and industry. In this thesis, the operational state Multiwavelength Analytical Ultracentrifuge (MWL-AUC) AUC is to be introduced. Consequently, three different applications of MWL-AUC to the aforementioned disciplines shall be presented. First of all, application of MWL-AUC to a biological system which is a mixture of proteins lgG, aldolase and BSA is presented. An application of MWL-AUC to a mass-produced industrial sample (β-carotene gelatin composite particles) which is manufactured by BASF AG, is presented. Finally, it is shown how MWL-AUC will impact on nano-particle science by investigating the quantum size effect of CdTe and its growth mechanism. In this thesis, mainly the relation between new technological developments and detector development for AUC is investigated. Pioneering results are obtained that indicate the possible direction to be followed for the future of AUC. As an example, each MWL-AUC data contains thousands of wavelengths. MWL-AUC data also contains spectral information at each radial point. Data can be separated to its single wavelength files and can be analyzed classically with existing software packages. All the existing software packages including Ultrascan, Sedfit, Sedanal can analyze only single wavelength data, so new extraordinary software developments are needed. As a first attempt, Emre Brookes and Borries Demeler have developed mutliwavelength module in order to analyze the MWL-AUC data. This module analyzes each wavelength separately and independently. We appreciate Emre Brookes and Borries Demeler for their important contribution to the development of the software. Unfortunately, this module requires huge amount of computer power and does not take into account the spectral information during the analysis. New software algorithms are needed which take into account the spectral information and analyze all wavelengths accordingly. We would like also invite the programmers of Ultrascan, Sedfit, Sedanal and the other programs, to develop new algorithms in this direction. / Die analytische Chemie versucht die chemische Zusammensetzung, chemische und physikalische Eigenschaften von biologischen oder künstlichen Materialien zu bestimmen. Mit der Entwicklung deren Methoden können genauere Informationen über die Umweltverschmutzung, das Ozonloch, Proteinfunktionen und Wechselwirkungen im menschlichen Körper erlangt werden. Es sind eine Vielzahl von analytischen Techniken vorhanden, die durch Verbesserungen in der Mikroelektronik, Mechanik, Informatik und Nanotechnologie einer markanten Entwicklung unterworfen wurden. In dieser Arbeit wurde versucht die Detektionskapazität der analytischen Ultrazentrifuge zu erhöhen. Die analytische Ultrazentrifuge (AUZ) ist eine gut bekannte, sehr leistungsstarke Trennungsmethode. AUZ benutzt die Zentrifugalkraft zum Trennen von Stoffen. Die Probe kann für die Messung gelöst oder in einer Flüssigkeit dispergiert werden. Makromoleküle, Proteine und kolloidale Systeme in Lösung können in einer AUZ Zelle zwischen 1000-60000 Rotationen pro Minute zentrifugiert werden, wie beispielsweise in der kommerziellen Beckmann AUZ. Die Rotationsbeschleunigung entspricht 73-262mal der Erdschwerebeschleunigung (= 9.81 m s-2) für eine radiale Position von 6.5 Zentimeter. Diese Kraft ist der Schlüsselfaktor für die Fähigkeit der AUZ sogar kleine Moleküle und Ionen zu trennen. Die Experimente wurden bei kontrollierter Rotationsgeschwindigkeit und Temperatur ausgeführt. Drei verschiedene, neue Detektoren wurden im Rahmen dieser Arbeit konstruiert und getestet. Diese Detektoren haben die analytischen Informationen sehr verbessert. Dies wurde für Proteine, halbleitende Nanopartikel sowie auch für industrielle Produkte gezeigt.

Analysis of the molecular speciation of organic compounds in solution is essential for the understanding of ionic complexation. The Raman spectroscopic technique was chosen for this purpose because it allows the identification of compounds in different states and it can give information about the molecular geometry from the analysis of the vibrational spectra. In this research the ionisation steps of relevant pharmaceutical material have been studied by means of potentiometry coupled with Raman spectroscopy; the protonation and deprotonation behaviour of the molecules were studied in different pH regions. The abundance of the different species in the Raman spectra of aqueous salicylic acid, paracetamol, citric acid and salicylaldoxime have been identified, characterised and confirmed by numerical treatment of the observed spectral data using a multiwavelength curve-fitting program. The non-destructive nature of the Raman spectroscopic technique and the success of the application of the multiwavelength curve-fitting program demonstrated in this work have offered a new dimension for the rapid identification and characterisation of pharmaceuticals in solution and have indicated the direction of further research. The work also covers the formation of novel cocrystal systems with pharmaceutically relevant materials. The existence of new cocrystals of salicylic acid-nicotinic acid, DLphenylalanine , 6-hydroxynicotinic acid, and 3,4-dihydroxybenzoic acid with oxalic acid have been identified from stoichiometric mixtures using combined techniques of Raman spectroscopy (dispersive and transmission TRS), X-ray powder diffraction and thermal analysis. Raman spectroscopy has been used to demonstrate a number of important aspects regarding the nature of the molecular interactions in the cocrystal. Cocrystals of II salicylic acid ¿ benzamide, citric acid-paracetamol and citric acid -benzamide have been identified with similar analytical approaches and structurally characterised in detail with single crystal X-ray diffraction. From these studies the high selectivity and direct micro sampling of Raman spectroscopy make it possible to identify spectral contributions from each chemical constituent by a peak wavenumber comparison of single-component spectra (API and guest individually) and the two- component sample material (API/guest), thus allowing a direct assessment of cocrystal formation to be made. Correlation of information from Raman spectra have been made to the X-ray diffraction and thermal analysis results. Transmission Raman Spectroscopy has been applied to the study cocrystals for the first time. Identification of new phases of analysis of the low wavenumber Raman bands is demonstrated to be a key advantage of the TRS technique. / Libyan government and Misurata University

Within the framework of the present work, an aerosol typing methodology applicable to both ground-based and spaceborne lidar systems has been developed. The novel aerosol typing scheme was developed based on the optimal estimation method (OEM) and allows the identification of up to four different aerosol components present in an aerosol mixture as well as the quantification of their contribution to the aerosol mixture in terms of relative volume. The four aerosol components considered in this typing scheme represent the most commonly observed aerosol particles in nature and are assumed to be physically separated from each other and, therefore, can create external mixtures. Two components represent fine-mode particles, absorbing (FSA) and less absorbing (FSNA), and the remaining two aerosol components represent coarse-mode particles, spherical (CS) and non-spherical (CNS). These components can adequately represent the most frequently observed aerosol types in the atmosphere: combustion- and pollution-related aerosol, sea salt and desert dust, respectively. The lidar-derived optical parameters used in this typing scheme are the lidar ratio and the particle linear depolarization ratio at two distinctive wavelengths (355 and 532 nm), the backscatter-related color ratio (for the wavelength pair of 532/1064 nm) and the extinction-related Ångström exponent (for the wavelength pair of 355/532 nm). These intensive optical properties can be combined in different ways making the methodology flexible, allowing thus its application to lidar systems with different configurations (e.g., single wavelength or multiwavelength). The functionality of the typing scheme was demonstrated by its application to case studies of known aerosol conditions as well as to cases of non-characterized aerosol load. The algorithm was also applied to a long-term dataset to provide a seasonal characterization of the aerosol situation over Haifa, Israel. It was shown that the OEM is an effective methodology that can be also applied for aerosol typing purposes, and that it can be used to support the ground-based validation efforts of EarthCARE's products and algorithms.

Made available in DSpace on 2016-03-15T19:37:34Z (GMT). No. of bitstreams: 1 Claudia Barros dos Santos.pdf: 3294972 bytes, checksum: b848d69d9c6a8c824e5142a4e3659d37 (MD5) Previous issue date: 2011-01-21 / In this work, we inserted two Arrayed Waveguide Gratings (AWGs), in an Erbium doped fiber ring cavity laser, where a single gain medium at room temperature can emit laser in multiples wavelengths, simultaneous and individualy controlled. The setup allowed us to check different functions in the ring cavity. Here we show emission in CW regime, Passive Mode-Locking, using Carbon Nanotubes as saturable absorbers and finally Active mode-locking at 7GHz, simultaneously. / Neste trabalho, fez-se a inserção de duas grades de difração matriciais com guias de onda, ou AWGs (Arrayed Waveguide Gratings) em uma cavidade laser de fibra dopada com Érbio, onde um único meio de ganho em temperatura ambiente pode gerar ação laser em múltiplos comprimentos de onda, com emissões simultâneas e controladas individualmente. A configuração utilizada permitiu o teste com diferentes regimes de operação simultâneos dentro da cavidade. Mostramos a possibilidade de emissão laser em regime CW, em regime de mode-locking passivo, com o uso de nanotubos de carbono como absorvedor saturável e, por último, em regime de modelocking ativo com uma freqüência de modulação de 7 GHz.

Made available in DSpace on 2016-03-15T19:37:35Z (GMT). No. of bitstreams: 1 Claudia Barros dos Santos.pdf: 3294972 bytes, checksum: b848d69d9c6a8c824e5142a4e3659d37 (MD5) Previous issue date: 2011-01-21 / In this work, we inserted two Arrayed Waveguide Gratings (AWGs), in an Erbium doped fiber ring cavity laser, where a single gain medium at room temperature can emit laser in multiples wavelengths, simultaneous and individualy controlled. The setup allowed us to check different functions in the ring cavity. Here we show emission in CW regime, Passive Mode-Locking, using Carbon Nanotubes as saturable absorbers and finally Active mode-locking at 7GHz, simultaneously. / Neste trabalho, fez-se a inserção de duas grades de difração matriciais com guias de onda, ou AWGs (Arrayed Waveguide Gratings) em uma cavidade laser de fibra dopada com Érbio, onde um único meio de ganho em temperatura ambiente pode gerar ação laser em múltiplos comprimentos de onda, com emissões simultâneas e controladas individualmente. A configuração utilizada permitiu o teste com diferentes regimes de operação simultâneos dentro da cavidade. Mostramos a possibilidade de emissão laser em regime CW, em regime de mode-locking passivo, com o uso de nanotubos de carbono como absorvedor saturável e, por último, em regime de modelocking ativo com uma freqüência de modulação de 7 GHz.

L'Univers est continûement le théâtre d'événements explosifs capables de relâcher une énorme quantité d'énergie sur des courtes échelles de temps. Ces sources transitoires comme les sursauts gamma, les supernovae ou les noyaux actifs de galaxie sont souvent associées à des objets extrêmes comme des étoiles à neutrons ou des trous noirs. De manière générale, ces sources émettent des radiations électromagnétiques dans une large bande spectrale voire sur la totalité du spectre pour les cas les plus extrêmes. Dès lors, une analyse multi-longueur d'onde est vitale pour étudier et comprendre la physique complexe de ces objets. De plus, au voisinage de ces sources, des particules (rayons cosmiques, RC) pourraient être efficacement accélérées jusqu'à des énergies très elevées dans des processus de chocs violents. L'interaction de ces RCs avec l'environnement peut conduire à la production d'un nombre significatif de neutrinos de hautes énergies. Par conséquent, l'étude des objets transitoires par le biais de l'astronomie neutrino offre la possibilité d'identifier enfin la nature des puissants accélérateurs cosmiques.Cette thèse est dédiée à l'étude de deux sources transitoires parmi les plus extrêmes dans l'Univers : les sursauts gamma (en anglais, Gamma-Ray Bursts : GRBs) détectés il y a ~ 50 ans et les sursauts radio (en anglais, Fast Radio Bursts : FRBs) fraîchement découverts il y a ~ 15 ans. Ces sources sont caractérisées par l'émission "prompte" d'un flash gamma (keV-MeV) durant de quelques ms à plusieurs secondes dans le cadre des GRBs et d'un flash intense en radio (GHz) durant quelques ms pour les FRBs. Dans le cas des GRBs une émission rémanente dite "afterglow" est observée dans une large gamme spectrale (X, visible et radio) alors que jusqu'à présent aucune autre contrepartie électromagnétique provenant d'un FRB n'a été découverte. Ces dernières années des modèles d'émission multi-longueur d'onde et multi-messager ont été développés afin d'expliquer ces 2 phénomènes. L'objectif majeur de ce travail de thèse est de tester ces modèles d'émission afin de contraindre la physique et la nature de ces deux objets. Pour cela, une analyse détaillée des propriétés physiques de l'émission afterglow des GRBs a été menée grâce à un large échantillon de données collectées ces 20 dernières années par diverses télescopes. Cette étude a permis de mettre en évidence les lacunes et les réussites du modèle GRB dit "standard" mais aussi les liens physiques subtils existant entre l'émission prompte des GRBs et leurs rémanences. Une recherche de signal neutrino en coïncidence avec les GRBs/FRBs a aussi été réalisée avec le télescope à neutrinos ANTARES. Les résultats sont décrits dans cette thèse ainsi que les contraintes apportées sur les processus d'accélération des particules durant ces phénomènes transitoires. Enfin, ce manuscrit rend compte des différents programmes d'observations innovants qui ont été engagés sur les télescopes optiques TAROT et Zadko et le télescope à neutrinos ANTARES afin de contraindre la nature des progéniteurs des GRBs/FRBs. / The Universe is continuously the scene of explosive events capable of releasing a tremendous amount of energy in short time scales. These transients like Gamma-Ray Bursts, Supernovae or Active Galactic Nuclei are often associated with extreme objects such as neutron stars or black holes. Generally, these sources emit light in a large spectral energy range and sometimes in the whole electromagnetic spectrum for the most extreme cases. Thus, a multi-wavelength analysis is crucial to study and understand the complex physical processes at work. Furthermore, in the vicinity of these sources, particles (cosmic-rays, CRs) could be efficiently accelerated up to very high energies by violent shock mecanisms. The interaction of these CRs with the surrounding environment may lead to a substantial production of high-energy neutrinos. Therefore, the study of the high-energy transient objects through neutrino astronomy offer the possibility to finally identify the nature of the powerful cosmic accelerators a hundred year after the discovery of the cosmic-rays.This thesis is dedicated to the study of two transient sources among the most extreme ones observed in the Universe: the Gamma-Ray Bursts (GRBs) detected ~ 50 years ago and the Fast Radio Bursts (FRBs) newly discovered ~ 15 years ago. These sources are characterised by the "prompt" emission of a gamma-ray flash (keV-MeV) lasting few ms up to few seconds for GRBs and an intense pulse of radio light (GHz) lasting few ms for FRBs. In the case of GRBs a late broadband afterglow emission is observed in X-rays/optical/radio domain while up to now no other electromagnetic counterpart has ever been detected in coincidence with any FRBs. These last years, many models predicting a multi-wavelength and a multi-messenger emission from these two phenomena have been developped. The main goal of this thesis work is to test these models in order to constrain the physics and the nature of the GRBs/FRBs. To do so, a detailed analysis on the physical properties of the GRB afterglow emission was made thanks to a large set of data collected these last 20 years by various facilities. The study reveals the major problems but also the successes encountered with the so-called "standard" GRB model. Subtle connections between the prompt and the afterglow emission are also discussed. In addition, a search for a neutrino signal from GRBs/FRBs was realised with the ANTARES neutrino telescope. The results are described in this thesis as well as the constraints on the particle acceleration mecanisms occuring during these transient phenomena.At last, this manuscript presents the different innovative observational programs realised in the optical domain with the TAROT and Zadko telescopes and in the astroparticle side with the ANTARES neutrino telescope in order to probe the nature of the GRBs/FRBs progenitors.