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.

In dieser Arbeit wird die digital-holographisch-interferometrische Zwei-Wellenlängen-Formerfassung sehr schnell bewegter Objekte behandelt und dafür unter Nutzung einer Ultrakurzpuls-Laserquelle mit der digitalen Einzelimpuls-Mehr-Wellenlängen-Holographie ein neuartigen Ansatz der digital-holographischen Aufnahme und Auswertung entwickelt. Mit der Entwicklung spezieller Methoden zur Formung der spektralen Signatur einer Ultrakurzpuls-Laserquelle hoher Leistungsdichte wurde zum ersten Mal die Voraussetzung für eine Zwei-Wellenlängen-Formerfassung hochdynamischer Objekte geschaffen. Die intrinsisch kurze Belichtungszeit unter einer Pikosekunde macht das Verfahren absolut stabil gegenüber Umwelteinflüssen. Für die simultane Aufnahme werden die spektral verschiedenen Hologramme mit einem eigens entwickelten Prinzip der Polarisationskodierung räumlich getrennt und zum ersten Mal mit zwei synchron laufenden Kameras gespeichert. Mit den in der digital-holographischen Einzelimpuls-Mehr-Wellenlängen-Interferometrie zusammengefassten numerischen Routinen zur Rekonstruktion und Phasenauswertung wird eine Zwei-Wellenlängen-Formerfassung mit mehreren Kameras möglich. In Anwendung des neuartigen Verfahrens an verschiedenen dynamischen Mikrosystemen konnte eine Genauigkeit von einem Zwanzigstel der erzeugten synthetischen Wellenlänge, bei der Auswertung der spektralen Differenzphase an Objekten in Reflexion erreicht werden. In einer digital-holographischen Hochgeschwindigkeitsformerfassung in Transmission wurden erstmals Bildfolgefrequenz von mehr als 0,4 kHz erreicht und der interferometrische Eindeutigkeitsbereich auf mehr als das 60-fache der optischen Wellenlänge ausgedehnt. Es wurden die Voraussetzungen für eine digitale Vier-Wellenlängen-Holographie geschaffen. Zukünftig wird eine Formerfassung mit einer Genauigkeit von 10nm über einen eindeutigen interferometrischen Bereich einiger 10 μm und die Untersuchungen von Prozessen auf einer Pikosekunden-Zeitskala möglich sein. / This work deals with the digital holographic interferometric two-wavelength contouring of very fast moving objects and develops with the digital single pulse multiwavelength holography a novel approach of digital holographic recording and analysis, using an ultrashort pulse laser source. The development of several methods to shape the spektral signature of an high power ultrashort pulse laser source provides the precondition for a two-wavelength contouring of highly dynamic objects for the first time. The intrinsically short exposure time shorter than a picosecond makes the system stable regarding external impacts. For the simultaneous recording the spektral different holograms are spatially separated in novel interferometric setups by the especially developed principle of polarization encoding and stored with two synchronized cameras for the first time. The digital holographic single pulse multi-wavelength interferometry combines the numeric routines of reconstruction and phase evaluation that make a two-wavelength contouring possible using more than one camera. The novel approach is successfully demonstrated on several dynamic microsystems. Evaluating the spectral phase difference for objects in reflection an accuracy of 2 μm, which corresponds to the twentieth of the realized synthetic wavelength, could be achieved. In a digital holographic high speed contouring in transmission a frame rate higher than 0,4 kHz was achieved for the first time and the interferometric range of unambiguity was extended larger than sixty times the optical wavelength. Furthermore, the developed digital holographic single pulse multi-wavelength interferometry is not limited to the evaluation of two wavelength. The principles of the method allow to perform digital four-wavelength holography. Future a contouring with an accuracy of 10nm over the unambiguous interferometric range of several 10 μm and the investigation of processes on a picosecond time scale will be possible.

The in ovo sexing of chicken eggs is a current task and a prerequisite to overcome the mass killing of male day-old chicks from laying lines. Although various methods have been developed and tested in recent years, practicable methods for sex determination are still missing which can be applicated in poultry hatcheries before the chicken embryo is capable of nociception and pain sensation. Optical spectroscopic methods enable an early determination of the sex. In this study, a novel method based on two-wavelength in ovo fluorescence excitation is described. More than 1600 eggs were examined. In ovo fluorescence was sequentially excited at 532 nm and 785 nm. The fluorescence intensities of the spectral regions behave inversely with respect to sex. It is shown that the observed sex-related differences in the fluorescence intensities are based on the embryonic hemoglobin synthesis. The accuracy of sex determination is 96% for both sexes. The hatching rate is not reduced compared to an equivalent reference group.

Das IceCube Neutrino-Observatorium ist ein am geografischen Südpol im Eis installierter Neutrinodetektor. In IceCube werden Neutrinos mit Tscherenkow-Strahlung von Sekundärteilchen aus Neutrino Interaktionen detektiert. Für den Nachfolgedetektor IceCube-Gen2, werden neue und verbesserte Lichtdetektoren gesucht. Die vorliegende Arbeit beschreibt die Entwicklung eines dieser Lichtdetektoren. Dieser basiert auf Wellenlängen schiebenden und Licht leitenden Technologien. Der Detektor mit dem Namen "Wavelength-shifting Optical Module" (WOM) verwendet eine transparente Röhre, mit wellenlängenschiebender Farbe, als passiver Photonendetektor. Das in der Wellenlänge verschobene Licht wird durch Totalreflexion, zu kleinen PMTs an beiden Enden geleitet. Die Auswahl dieses Designs reduziert die Kosten und verbessert das Signal-Rausch-Verhältnis wesentlich, möglicherweise können mit dieser Lösung extragalaktische Supernova in zukünftigen Detektoren beobachtet werden. Als eine Kernkomponente wird die wellenlängenschiebende Röhre ausführlich untersucht. Verschiedene Messaufbauten und Auswertungsmethoden werden entwickelt, um diese im Anschluss zu untersuchen und zu bewerten. Iterative Verbesserungen der Materialien und des Farbauftrageverfahren als auch Messmethoden, resultieren in einer kombinierten Einfang-, Wellenlängenschiebe- und Transporteffizienz von 28,1 +/- 5,4 % der Röhre. Ein Model zur Beschreibung des Lichtverhaltens in der Röhre wird entwickelt um eine Diskrepanz zwischen Theorie und Messung zu untersuchen. Die Kombination zwischen Messung und Model, bestätigt die Aussagekraft des Models und zeigt, dass ein Großteil der Verluste beim Lichttransport zustande kommen. Darüber hinaus werden die physikalischen Eigenschaften des WOM in die IceCube Simulationsumgebung eingebaut. Der Vergleich zu einem Konkurrenzmodul zeigt eine Überlegenheit des WOM um den Faktor 1,05 +/- 0,07. Es werden Vorschläge und Ausblicke für Verbesserungen der Leistungsfähigkeit des WOMs gegeben. / The IceCube Neutrino Observatory is an in ice neutrino detector located at the geographic South Pole. In IceCube neutrinos are detected via Cherenkov light produced by secondary particles in neutrino interactions. For the upgraded detector IceCube-Gen2, new and improved light detectors are sought-after. This work describes the development of one of those light detectors based on a novel combination of wavelength-shifting and light-guiding technology. The detector named the Wavelength-shifting Optical Module (WOM) utilizes a large transparent tube, coated with wavelength-shifting paint as a passive photon detector. The wavelength-shifted light is guided via total internal reflection towards small active light detectors, at each end of the tube. This design reduces costs and improves the signal to noise ratio significantly, thereby potentially enabling extragalactic supernova detections in future detectors. As a core component, the wavelength-shifting tube is extensively investigated. Different measurement setups and evaluation techniques are developed and investigated. Iterative improvement of materials and coating techniques as well as measurement methods currently result in a combined photon capture, shift and transport efficiency of 28.1 +/- 5.4 % for the tube. Those results contrast the theoretical maximum of 74.5 %. A model is developed to describe the light propagation and loss processes in the tube and to understand the discrepancies between theory and measurement. The combination of the measurements with the model, validate the descriptive qualities of the model and show that most of the light is lost during the light propagation in the tube. Additionally, the physical properties of the WOM are included in the IceCube simulation framework. A comparison to a competing module showed that the WOM outperforms by a factor of 1.05 +/- 0.07 in photon detection numbers. Where applicable, suggestions and outlooks are given to enhance the performance of the WOM.

This work relates to the remote sensing technologies to determine wind velocity and its related phenomena. Laser-based pulsed systems with heterodyne detection allow a very sensitive measurement of particles’ velocity in the atmosphere. The heterodyne detection theory and the essential principles and properties of this technology are presented. An all-fiber coherent Doppler lidar system in master-oscillator power-amplifier design with a novel multi-wavelength channel configuration was developed and introduced, characterized, and validated in this work. The fiber amplifier is a crucial component of multi-channel all-fiber systems and is therefore discussed with special emphasis on non-linear effects and pulse distortions. A novel feedback controlled pulse-shaping unit was developed to control pulse energy and the shape of the amplified pulses of each channel. Each and every wavelength-channel creates an independent speckle pattern, which is demonstrated by hard targets with diffuse character and atmospheric single-shot measurements, and compared with theoretical results. Utilizing four channels, the precision of the measured wind velocity can be improved and this feature is demonstrated with measured atmospheric return signals. A correlation technique is presented, which enables the enhancement of the SNR at higher backscattered powers by utilizing multiple channels. The multi-wavelength system was designed to work with four wavelength-channels, which also allow measurements along the same line-of-sight with different channel configurations. These channels can be different in pulse shape, pulse length, and pulse repetition frequency. In this work, the resulting benefits of using multiple channels are shown with atmospheric measurements and scientific discussion of these is presented.:1. Introduction to the subject 2. Introduction to pulsed coherent Doppler lidar 3. Methodology: Multi-channel coherent Doppler lidar system 4. Erbium-doped fiber amplifier in a multi-channel lidar 5. Correlation properties of a multi-channel lidar system 6. Benefits of a multi-channel coherent Doppler lidar 7. Conclusions A. Wind measurement examples B. Range gate weighting function for different pulse shapes / Diese Arbeit befasst sich mit der Fernerkundungstechnologie zur Erfassung der Windgeschwindigkeit und den damit verbundenen Phänomenen. Die heterodyne Detektion der laserbasierten gepulsten Systeme ermöglicht eine sehr sensitive Messung von Partikelgeschwindigkeiten in der Atmosphäre. Sowohl die Theorie der heterodynen Detektion als auch die wesentlichen Eigenschaften und Grundsätze werden in dieser Arbeit diskutiert. Ein neuartiges faserbasiertes kohärentes Doppler Mehrkanal-Lidarsystem wurde entwickelt, vorgestellt, charakterisiert und validiert. Dabei ist der faserbasierte Verstärker eine wesentliche Komponente in diesem System und wird somit in Bezug auf Nichtlinearitäten und Impulsverzerrungen besonders analysiert. Eine neuartige geregelte Impulsformung wurde entwickelt, um die Impulsenergie und die Impulsform unabhängig für jeden Kanal zu stabilisieren. Jeder einzelne Kanal erzeugt ein unabhängiges Specklebild, welches durch Messungen am festen Ziel und durch atmosphärische Messungen präsentiert und mit theoretischen Berechnungen verglichen wird. Somit wird unter der Verwendung aller vier Kanäle des Systems die Geschwindigkeitsgenauigkeit erhöht, welches durch atmosphärische Messungen dargelegt wird. Zudem wird eine Korrelationstechnik vorgestellt, die das SNR bei höherer Rückstreuintensität weiter erhöht. Das Mehr-Wellenlängensystem wurde für vier Kanäle ausgelegt, welches eine Beobachtung der Atmosphäre entlang der Sichtlinie unter sonst gleichen Bedingungen ermöglicht. Diese Kanäle können sich sowohl in der Impulsform, in der Impulslänge als auch in der Pulsfolgefrequenz unterscheiden. Diese Arbeit zeigt die wesentlichen Vorteile eines solchen Mehrkanal-Lidarsystems mit atmosphärischen Messungen und wissenschaftlichen Ausarbeitungen.:1. Introduction to the subject 2. Introduction to pulsed coherent Doppler lidar 3. Methodology: Multi-channel coherent Doppler lidar system 4. Erbium-doped fiber amplifier in a multi-channel lidar 5. Correlation properties of a multi-channel lidar system 6. Benefits of a multi-channel coherent Doppler lidar 7. Conclusions A. Wind measurement examples B. Range gate weighting function for different pulse shapes

Wavelength-discriminating systems typically consist of heavy benchtop-based instruments, comprising diffractive optics, moving parts, and adjacent detectors. For simple wavelength measurements, such as lab-on-chip light source calibration or laser wavelength tracking, which do not require polychromatic analysis and cannot handle bulky spectroscopy instruments, lightweight, easy-to-process, and flexible single-pixel devices are attracting increasing attention. Here, a device is proposed for monotonously transforming wavelength information into the time domain with room-temperature phosphorescence at the heart of its functionality, which demonstrates a resolution down to 1 nm and below. It is solution-processed from a single host–guest system comprising organic room-temperature phosphors and colloidal quantum dots. The share of excited triplet states within the photoluminescent layer is dependent on the excitation wavelength and determines the afterglow intensity of the film, which is tracked by a simple photodetector. Finally, an all-organic thin-film wavelength sensor and two applications are demonstrated where this novel measurement concept successfully replaces a full spectrometer.