La scintillation organique correspond au phénomène d’émission de lumière par un matériau moléculaire à la suite de l’excitation de celui-ci par un rayonnement externe d’énergie donnée. Lors de l’interaction, le dépôt d’énergie induit des transitions électroniques peuplant des états dont la plupart se désexcite de manière non radiative, à l’exception d’une, entre le premier état électronique singulet et l’état fondamental de la molécule. Lors de cette relaxation, un photon de fluorescence est émis. Cette émission a deux origines : i) l’excitation directe par le rayonnement primaire et les électrons secondaires ; elle donne lieu à une émission dite rapide ou prompte ; ii) l’ionisation par le rayonnement primaire et les électrons secondaires ; elle donne lieu à une émission dite lente ou différée. Ce travail de recherche fondamentale, à la fois théorique et expérimental, fait l’analyse de toutes les étapes du processus, de l’interaction primaire à l’émission de fluorescence, de manière à relier la dose déposée à la quantité de lumière émise, à des fins d’applications en dosimétrie médicale. Il repose sur la mesure des déclins de fluorescence de deux molécules modèles, l’anthracène et le paraterphényle, excitées par un flux continu de rayons X, et la séparation des contributions rapide et lente de la lumière émise, aux énergies médicales. Une modélisation analytique des processus physiques conduisant à l’émission de lumière, au regard de la dose déposée, a ensuite été effectuée, faisant apparaître de nombreux résultats originaux. Dans un premier temps, un dispositif expérimental original a été développé, basé sur la technique TCSPC (Time-Correlated Single Photon Counting), afin de pouvoir mesurer des déclins temporels de fluorescence en résolution nanoseconde et sous flux d’irradiation continu. Dans un second temps, nous avons développé une nouvelle approche mathématique permettant d’extraire finement les composantes rapides et lentes du signal. L’analyse des résultats a montré, pour la première fois, l’existence d’un rapport R constant et uniquement fonction du matériau, entre les rendements d’excitation et d’ionisation. Le caractère constant de ce rapport ne peut être attribué qu’à un mécanisme d’autoionisation moléculaire au sein d’un matériau se comportant intrinsèquement comme une chambre d’ionisation proportionnelle pour l’ionisation secondaire de basse énergie. Ceci est en accord total avec la linéarité observée entre l’intensité totale de lumière différée (ionisation) et la dose mesurée par une chambre d’ionisation proportionnelle. Une étude plus approfondie des mécanismes d’excitation, au regard du rapport R, a également permis de montrer, pour la première fois, une proportionnalité directe entre l’intensité totale de la lumière prompte et le dépôt d’une dose que nous avons baptisé dose d’excitation. Cette dose a été observée comme étant de 4 à 14 fois supérieure à celle mesurée par une chambre d’ionisation. Ce résultat original majeur devra impérativement conduire à des études futures afin de mieux comprendre les dégâts infligés à la matière organique et biologique par les excitations. / Organic scintillation is the emission of light by an organic scintillator when irradiated by an external source of radiation depositing enough energy to excite the molecule. Electronic states are populated by the electronic transitions generated by the deposited energy. The states de-excite through radiationless transitions, except for one, the transition between the first electronic state and the ground state where a photon of fluorescence is emitted. This light has two different origins: i) direct excitation caused by primary radiation or secondary electrons which leads to an emission knows as prompt; ii) ionization caused by primary radiation or secondary electrons generate what is known as the delayed component. This fundamental research was based on both theoretical and experimental work. We studied all the different processes in organic scintillation, from the interaction between the incident radiation and matter to the emission of light in order to find the relationship between fluorescence and the deposited dose, to the application to medical dosimetry. Two well known organic scintillators, anthracene and p-terphenyl, were excited using an X-ray source set at typical medical imaging parameters. The light emitted was acquired and an analytical model was used to describe the different processes that led to light emission revealing interesting new results.An experimental setup, based on the Time Correlated Single Photon Counting (TCSPC) technique, was developed to acquire fluorescence decay curves with nanosecond resolution using a continuous X-ray source. Afterwards, these curves were analyzed using an innovative mathematical approach in order to determine the prompt and delayed components.Results showed the ratio, defined as R, between the prompt and delayed components of fluorescence was constant and independent of the energy of the incident X-rays and that the response of the delayed component of fluorescence was linear to an ionization chamber. These observations were explained by considering that the only process taking place within the molecule after excitation was autoionization. Hence, the response of organic scintillator was the same as the one of an ionization chamber. Furthermore, due to the constant ration R, the response of prompt component of fluorescence was linear to the ionization chamber as well. This was the first time this behavior was observed and we referred to it as excitation dose. This dose was between 4 and 14 times bigger than the one measured with the ionization chamber. These original results suggested that energy is deposited mainly through excitation processes, suggesting the need for further studies to better understand the damage caused by excitation to the living.

Ein Ziel der Arbeit war die Entwicklung eines Verfahrens zur 3D-Positionierung auf Basis elektrooptischer Pseudostreckenmessung. Ein zweites Ziel war die Entwicklung eines Reflexgoniometers zur zweiachsigen Winkelmessung. Im Rahmen der Arbeit wurden die Grundlagen zur instrumentellen Umsetzung beider Verfahren erarbeitet, die Genauigkeitspotentiale ermittelt und mögliche Anwendungen für die einzelnen Schlüsseltechnologien und deren Kombination abgeleitet. In einer Prototyp-Entwicklung wurden Vorschläge für die wesentlichen Funktionselemente des räumlichen Weg- bzw. Winkelmesssystems gemacht. Hierzu gehören das kardanisch aufgehängte Etalon, die temperaturstabilisierte Laseroptik und die temperaturkalibrierte CCD der Winkelmesseinheit (Reflexgoniometer), die Systeme zur Erzeugung und Detektion ultrakurzer Laserpulse, eine elastische Optik, Möglichkeiten zur Formung des transversalen Strahlprofils, das TCSPC-System und die zur Auswertung und Kalibrierung notwendigen Algorithmen, wie z.B. die Autokollimation eines Lasers. ("Veröffentlicht von der Deutschen Geodätischen Kommission Reihe C (Dissertationen) unter der Nummer C 614 (München 2008; ISBN 3 7696 5053 0; 2; 144 S.") / One goal of the thesis was the development of a method for three-dimensional positioning based on electro-optical measurement of pseudo ranges. Another goal was the development of a reflex goniometer for biaxial angle measurement. Within the scope of this thesis the basics for the instrumental realisation of both methods were developed, the accuracy potentials were determined and possible applications for the separate key technologies and their combination were deduced. In a prototype development proposals were made for the main functional elements of the spatial distance and angle measurement systems. These include the gimbal mounted etalon, the temperature stabilised laser optics and the temperature calibrated CCD of the angle measurement device (reflex goniometer), the systems for creation and detection of ultrashort laser pulses, an elastic optical device, possibilities of transversal beam shaping, the TCSPC system and the algorithms which are necessary for analysis and calibration, e.g. the autocollimation of a laser.

<div><p><br></p></div><p>This study makes use of a Time-Resolved Raman Spectroscopy (TRRS) system developed in the Purdue Civil Engineering spectroscopy laboratory to advance technology critical to enable field deployment of Raman spectroscopic systems, with a primary focus on developing solutions to overcome two specific barriers to Raman analysis in the natural environment: (1) obtaining Raman spectra of chemical compounds at field-relevant concentrations, and (2) realizing economical spatial monitoring. To inform both streams of activity, this work first explores the role of component choice and apparatus design on Raman system output. A component-level Raman system transfer function is developed in terms of intensity, wavelength, and time which yields detailed insight into system performance that greatly exceeds traditional single “system factor” treatments of apparatus effects. The modelling frame provided by the transfer function is universally applicable in that it is inclusive of the majority of component choices that may be encountered in any open-path or closed-path Raman system, and is likely to be valuable in efforts to assess the performance benefits and limitations of system designs, modify or tailor apparatus layouts, facilitate experiment design, and compare results obtained on different systems. </p><p><br></p><p>The system characterization offered by the transfer function is then employed to develop a multi-photon counting algorithm realized through digital signal processing (DSP) which captures photon arrivals traditionally ignored in conventional counting methods. This approach increases acquired Raman intensity for any given analyte by using detector output voltage or a voltage-time product as an energy proxy – an approach that is likey broadly applicable to any spectroscopic techniques employing detectors that make use of the photoelectric effect. In experiments carried out on analytes (nitrate, isopropanol, and rhodamine 6G) in aqueous solutions, enhanced observations enabled by the multi-photon counting algorithm are shown to increase observed Raman intensities of low Raman-yield solutions 2.0-3.1-fold compared to single-threshold analysis, and also extend the upper observation limit of strong Raman-yield solutions that would traditionally saturate detectors using a binary photon counting scheme. Notably, the improved performance offered by the multi-photon counting algorithm is realized through comparison of multi-photon and conventional counting algorithms applied to the same data in a post-processing exercise, thus eliminating any effects of test-to-test variation on results, and highlighting the ability to employ the developed counting approach without modification of traditional systems.</p><p><br></p><p>Additional insights from the system transfer function are also used to inform exploration of a novel approach to enable spatial environmental monitoring via Raman spectroscopy by combining fiber optics, optical switch technology, and the Raman system prototype. Tests designed to evaluate the system configured as a multiplexed optically switched fiber optic network demonstrate the potential to deliver excitation and collect Raman scattering from different desired monitoring locations with a sole excitation source and a single detector over substantial distances. Using nitrate as an example compound of interest, it is demonstrated that the system has a detection limit of 5 ppm within approximately 1.5 meters, which increases to 15 ppm at 100 m, and 38 ppm at 200 m. Modelling informed using the developed system transfer function highlights that improving the prototype by eliminating fiber connectors and making use of commercially available visible-light optimized fiber can substantially extend the range of the system, offering a 15-ppm nitrate detection limit at 2100 m. As increases in laser power, testing time, and collection optic efficiency are all also straightforward and viable, the prototype demonstrates realistic potential to achieve field relevant detection sensitivity over great distance.</p><p><br></p><p>As a final demonstration of system potential, a set of experiments on aqueous nitrate solutions is performed to understand the influence of turbidity, fluorescence, optics size, and varied raw data integration lengths on Raman observations. Results demonstrate that cumulative advances in the TRRS system establish a new generation of Raman spectroscopic sensing amenable to long-term environmental monitoring over significant spatial extent in complex in-situ conditions. Specific advances made herein include enhanced power delivery and scattered light collection informed by the system transfer function, increases in sensitivity from multi-photon counting, and incorporation of optical multiplexing. Overall, the Time-Resolved Raman Spectroscopic System (TRRS) now offers a set of capabilities that bring in-field deployment within practical reach.</p>

Ein Ziel der Arbeit war die Entwicklung eines Verfahrens zur 3D-Positionierung auf Basis elektrooptischer Pseudostreckenmessung. Ein zweites Ziel war die Entwicklung eines Reflexgoniometers zur zweiachsigen Winkelmessung. Im Rahmen der Arbeit wurden die Grundlagen zur instrumentellen Umsetzung beider Verfahren erarbeitet, die Genauigkeitspotentiale ermittelt und mögliche Anwendungen für die einzelnen Schlüsseltechnologien und deren Kombination abgeleitet. In einer Prototyp-Entwicklung wurden Vorschläge für die wesentlichen Funktionselemente des räumlichen Weg- bzw. Winkelmesssystems gemacht. Hierzu gehören das kardanisch aufgehängte Etalon, die temperaturstabilisierte Laseroptik und die temperaturkalibrierte CCD der Winkelmesseinheit (Reflexgoniometer), die Systeme zur Erzeugung und Detektion ultrakurzer Laserpulse, eine elastische Optik, Möglichkeiten zur Formung des transversalen Strahlprofils, das TCSPC-System und die zur Auswertung und Kalibrierung notwendigen Algorithmen, wie z.B. die Autokollimation eines Lasers. (&amp;quot;Veröffentlicht von der Deutschen Geodätischen Kommission Reihe C (Dissertationen) unter der Nummer C 614 (München 2008; ISBN 3 7696 5053 0; 2; 144 S.&amp;quot;) / One goal of the thesis was the development of a method for three-dimensional positioning based on electro-optical measurement of pseudo ranges. Another goal was the development of a reflex goniometer for biaxial angle measurement. Within the scope of this thesis the basics for the instrumental realisation of both methods were developed, the accuracy potentials were determined and possible applications for the separate key technologies and their combination were deduced. In a prototype development proposals were made for the main functional elements of the spatial distance and angle measurement systems. These include the gimbal mounted etalon, the temperature stabilised laser optics and the temperature calibrated CCD of the angle measurement device (reflex goniometer), the systems for creation and detection of ultrashort laser pulses, an elastic optical device, possibilities of transversal beam shaping, the TCSPC system and the algorithms which are necessary for analysis and calibration, e.g. the autocollimation of a laser.

<p>We have studied fundamental aspects of time-resolved two-photon excited fluorescence depolarisation. The thesis presents experimental as well as theoretical progress. We show that a multi-photon induced instrumental response function obtained from a suspension of gold nanoparticles is appropriate for the analysis of two-photon excited fluorescence decays obtained using time-correlated single photon counting detection. Theoretical expressions have been derived for the fluorescence anisotropy decay obtained upon two-photon excitation of various molecular systems in liquid solutions: a) an anisotropic rigid rotor that undergoes rotational diffusion in the presence of ultrafast unresolved restricted reorientations, e.g. librations. b) a molecular group covalently attached to a stationary macromolecule, and undergoing local reorientation in a uniaxial ordering potential. A new approach to the analysis of two-photon excited fluorescence depolarisation experiments was developed, which combines data obtained by using linearly and circularly polarised excitation light, in a global manner. In the analysis, knowledge about unresolved reorientations was obtained from one-photon excitation studies of the corresponding systems. By means of this procedure it has been possible to obtain quantitative information about the molecular two-photon absorption tensor for perylene and two of its derivatives. Thereby the symmetry of the final excited and intermediate vibronic states could be assigned. The analysis reveals that the two-photon transition studied with the 800 nm laser exhibits mixed character. An important finding from the experiments was that the two-photon absorption tensor appears to be solvent dependent. Furthermore, the thesis presents the first theoretical treatment of two-photon excited donor-donor energy migration in the presence of molecular reorientation and which applies the extended Förster theory. Explicit expressions for molecules that belong to the point groups D_2h, D₂ and C_2v are given. Preliminary experiments are finally also reported on a two-photon excited donor-donor energy migration system consisting of a bisanthryl-bisteroid. </p>

This dissertation attempts to understand how energy transfer in a molecular wire and a spherical organic assembly are affected by molecular structure. The molecular wire is a DNA-hybrid structure composed of a strand of thymine bases appended by a cyanine dye. Hydrogen bonded to each base is a naphthalene-derivative molecule. Using time-integrated photoluminescence and time-correlated single photon counting measurements, energy transfer from the naphthalene donors to the cyanine acceptors was confirmed, and its dependence on temperature and DNA-template length investigated. Donor-thymine bonding was disrupted at temperatures above about 25 degrees Celcius resulting in poor donor template decoration and low rates of energy transfer. Increasing numbers of donors attach to the scaffold, forming an orderly array, as the template length increases due to the stabilising effects of the donor-donor pi-stacking interactions. Conversely, modelled energy transfer rates fall as the scaffold length increases because of the longer donor-acceptor distances involved. Therefore, the energy transfer rate was greatest for a template built from 30 thymines. The spherical organic assemblies (nanoparticles) are formed by fast injection of a small volume of molecularly dissolved fluorene-derivative amphiphilic molecules into a polar solvent. The amphiphilic molecules contained either a naphthalene (donor) or a benzothiadiazole (acceptor) core. The donor-acceptor mixed nanoparticles resemble an amorphous polymer film and were modelled as such using the Foerster resonance energy transfer theory. The Foerster radii extracted from the measurements depends intricately on the donor-acceptor spectral overlap and distance. The latter effect was controlled by the stacking interactions between the molecules. Altering the morphology of the structural units is the key to optimising energy transfer in molecular structures. To achieve efficient organic molecule-based devices, the importance of this property needs to be fully appreciated and effectively exploited.

Ma thèse de doctorat s’inscrit à l’interface entre la physique expérimentale et la biologie. Ce travail a été développé au sein de l’équipe imXgam du CPPM, qui a construit un prototype de micro-tomographie pour le suivi non-invasif du petit animal, équipé d’une caméra à pixels hybrides XPAD3 fonctionnant en mode comptage de rayons X. Le comptage de rayons X rendu possible par la technologie des pixels hybrides, permet de s’affranchir du bruit électronique et d’augmenter ainsi la détectabilité des tissus faiblement contrastés. Elle présente de plus la capacité d'appliquer à chaque pixel un seuil de détection en énergie permettant d’accéder à l’information spectrale des rayons X détectés et ouvre la voie au développement d’une méthode d’imagerie spectrale dite au K-edge, qui permet de différencier des agents de contraste particuliers. La caméra XPAD3 développée avec un capteur en Si présente une efficacité de détection qui limite son utilisation pour l’imagerie du vivant. Une caméra XPAD3 avec une meilleure efficacité au delà de 25 keV a été assemblée avec des capteurs en CdTe. Dans un premier temps, nous avons effectué une comparaison des caméras XPAD3/Si et XPAD3/CdTe en imagerie d’absorption standard et en imagerie au K-edge. Nous avons ensuite, en collaboration avec des biologistes de l’IBDM, assuré le suivi quantitatif et in vivo sur plusieurs mois, du développement de tumeurs hépatiques chez un modèle spécifique de souris et de l’efficacité d’un traitement ciblant les cellules tumorales. Enfin, nous avons développé un protocole d’acquisition spectrale à faible dose pour réaliser une tomographie spectrale in vivo d’un foie de souris en exploitant le K-edge du baryum. / My PhD thesis is at the interface between experimental physics and biology. This work has been developed within the imXgam team at CPPM, which has built a micro-computed tomography prototype for the non-invasive longitudinal monitoring of small animal, equipped with the XPAD3 hybrid pixel camera that operates in X-ray photon counting mode. X-ray photon counting that has been made possible by hybrid pixels, allows to free images from the electronic noise and thus to increase detectability of weakly contrasted tissues. Moreover, it provides the possibility to set an energy threshold for each pixel that allows to accessing spectral information on the detected X-rays and paving the way to the development of a spectral imaging modality also named K-edge imaging, which allows to differentiate selected contrast agents. Actually, the XPAD3 camera developed with a Si sensor presents a low detective efficiency that limits its use for biomedical imaging. A XPAD3 camera with a better efficiency above 25 keV has been assembled with high-Z CdTe sensors. Firstly, we have performed a comparison of XPAD3/Si and XPAD3/CdTe cameras for standard absorption CT and K-edge imaging. Then, in collaboration with a team of biologists from IBDM, we have carried out the quantitative and in vivo follow-up of hepatic tumour development in a specific mouse model over several months, and of the effectiveness of a treatment targeting these tumour cells. Finally, we have developed a protocol for low dose acquisition of spectral data to realize an in vivo spectral tomography of a mouse liver using the barium spectral signature.

La tomographie (CT) aux rayons X joue un rôle important dans l'imagerie non invasive depuis son introduction. Au cours des dernières années, de nombreuses avancées technologiques en tomographie par rayons X ont été observées, notamment la CT spectrale, qui utilise un détecteur à comptage de photons (PCD) pour discriminer les photons transmis correspondant à des bandes d'énergie sélectionnées afin d'obtenir une information spectrale. La CT spectrale permet de surmonter de nombreuses limitations des techniques précédentes et ouvre de nombreuses applications nouvelles, parmi lesquelles la décomposition quantitative des matériaux est le sujet le plus étudié. Un certain nombre de méthodes de décomposition des matériaux ont été rapportées et différents systèmes expérimentaux sont en cours de développement pour la CT spectrale. Selon le type de données sur lequel l'étape de décomposition fonctionne, nous avons les méthodes du domaine des projections (décomposition avant reconstruction) et les méthodes du domaine de l'image reconstruite (décomposition après reconstruction). La décomposition couramment utilisée est basée sur le critère des moindres carrés, nommée proj-LS et méthode ima-LS. Cependant, le problème inverse de la décomposition du matériau est généralement mal posé et les mesures du CT spectral aux rayons X souffrent de bruits de comptage de photons de Poisson. Le critère des moindres carrés peut conduire à un surajustement des données de mesure bruitées. Dans le présent travail, nous avons proposé un critère de moindre log-carré pour la méthode du domaine de projection afin de minimiser les erreurs sur le coefficient d'atténuation linéaire: méthode proj-LLS. De plus, pour réduire l'effet du bruit et lisser les images, nous avons proposé d'ajouter un terme de régularisation par patch pour pénaliser la somme des variations au carré dans chaque zone pour les décompositions des deux domaines, nommées proj-PR-LLS et ima -PR-LS méthode. Les performances des différentes méthodes ont été évaluées par des études de simulation avec des fantômes spécifiques pour différentes applications: (1) Application médicale: identification de l'iode et du calcium. Les résultats de la décomposition des méthodes proposées montrent que le calcium et l'iode peuvent être bien séparés et quantifiés par rapport aux tissus mous. (2) Application industrielle: tri des plastiques avec ou sans retardateur de flamme. Les résultats montrent que 3 types de matériaux ABS avec différents retardateurs de flamme peuvent être séparés lorsque l'épaisseur de l'échantillon est favorable. Enfin, nous avons simulé l'imagerie par CT spectrale avec un fantôme de PMMA rempli de solutions de Fe, Ca et K. Différents paramètres d'acquisition, c'est-à-dire le facteur d'exposition et le nombre de bandes d'énergie, ont été simulés pour étudier leur influence sur la performance de décomposition pour la détermination du fer. / X-ray computed tomography (X-ray CT) plays an important part in non-invasive imaging since its introduction. During the past few years, numerous technological advances in X-ray CT have been observed, including spectral CT, which uses photon counting detectors (PCDs) to discriminate transmitted photons corresponding to selected energy bins in order to obtain spectral information with one single acquisition. Spectral CT enables us to overcome many limitations of the conventional CT techniques and opens up many new application possibilities, among which quantitative material decomposition is the hottest topic. A number of material decomposition methods have been reported and different experimental systems are under development for spectral CT. According to the type of data on which the decomposition step operates, we have projection domain method (decomposition before reconstruction) and image domain method (decomposition after reconstruction). The commonly used decomposition is based on least square criterion, named proj-LS and ima-LS method. However, the inverse problem of material decomposition is usually ill-posed and the X-ray spectral CT measurements suffer from Poisson photon counting noise. The standard LS criterion can lead to overfitting to the noisy measurement data. In the present work, we have proposed a least log-squares criterion for projection domain method to minimize the errors on linear attenuation coefficient: proj-LLS method. Furthermore, to reduce the effect of noise and enforce smoothness, we have proposed to add a patchwise regularization term to penalize the sum of the square variations within each patch for both projection domain and image domain decomposition, named proj-PR-LLS and ima-PR-LS method. The performances of the different methods were evaluated by spectral CT simulation studies with specific phantoms for different applications: (1) Medical application: iodine and calcium identification. The decomposition results of the proposed methods show that calcium and iodine can be well separated and quantified from soft tissues. (2) Industrial application: ABS-flame retardants (FR) plastic sorting. Results show that 3 kinds of ABS materials with different flame retardants can be separated when the sample thickness is favorable. Meanwhile, we simulated spectral CT imaging with a PMMA phantom filled with Fe, Ca and K solutions. Different acquisition parameters, i.e. exposure factor and number of energy bins were simulated to investigate their influence on the performance of the proposed methods for iron determination.

<p>This project investigates a laser radar system. The system is based on the principles of time-correlated single photon counting, and by measuring the times-of-flight of reflected photons it can find range profiles and perform three-dimensional imaging of scenes. Because of the photon counting technique the resolution and precision that the system can achieve is very high compared to analog systems. These properties make the system interesting for many military applications. For example, the system can be used to interrogate non-cooperative targets at a safe distance in order to gather intelligence. However, signal processing is needed in order to extract the information from the data acquired by the system. This project focuses on the analysis of different signal processing methods.</p><p>The Wiener filter and the Richardson-Lucy algorithm are used to deconvolve the data acquired by the photon counting system. In order to find the positions of potential targets different approaches of non-linear least squares methods are tested, as well as a more unconventional method called ESPRIT. The methods are evaluated based on their ability to resolve two targets separated by some known distance and the accuracy with which they calculate the position of a single target, as well as their robustness to noise and their computational burden.</p><p>Results show that fitting a curve made of a linear combination of asymmetric super-Gaussians to the data by a method of non-linear least squares manages to accurately resolve targets separated by 1.75 cm, which is the best result of all the methods tested. The accuracy for finding the position of a single target is similar between the methods but ESPRIT has a much faster computation time.</p>

We have studied fundamental aspects of time-resolved two-photon excited fluorescence depolarisation. The thesis presents experimental as well as theoretical progress. We show that a multi-photon induced instrumental response function obtained from a suspension of gold nanoparticles is appropriate for the analysis of two-photon excited fluorescence decays obtained using time-correlated single photon counting detection. Theoretical expressions have been derived for the fluorescence anisotropy decay obtained upon two-photon excitation of various molecular systems in liquid solutions: a) an anisotropic rigid rotor that undergoes rotational diffusion in the presence of ultrafast unresolved restricted reorientations, e.g. librations. b) a molecular group covalently attached to a stationary macromolecule, and undergoing local reorientation in a uniaxial ordering potential. A new approach to the analysis of two-photon excited fluorescence depolarisation experiments was developed, which combines data obtained by using linearly and circularly polarised excitation light, in a global manner. In the analysis, knowledge about unresolved reorientations was obtained from one-photon excitation studies of the corresponding systems. By means of this procedure it has been possible to obtain quantitative information about the molecular two-photon absorption tensor for perylene and two of its derivatives. Thereby the symmetry of the final excited and intermediate vibronic states could be assigned. The analysis reveals that the two-photon transition studied with the 800 nm laser exhibits mixed character. An important finding from the experiments was that the two-photon absorption tensor appears to be solvent dependent. Furthermore, the thesis presents the first theoretical treatment of two-photon excited donor-donor energy migration in the presence of molecular reorientation and which applies the extended Förster theory. Explicit expressions for molecules that belong to the point groups D2h, D2 and C2v are given. Preliminary experiments are finally also reported on a two-photon excited donor-donor energy migration system consisting of a bisanthryl-bisteroid.