The resonant absorption, emission, and scattering of light are the fundamental optical processes that have been used both to probe matter and to manipulate light itself. In the last decade there has been essential progress in coherent control of both linear and nonlinear optical responses based on resonant excitation of atomic coherence in multilevel quantum systems. Some interesting and useful phenomena, resulting from coherent control of absorption and the group index, such as electromagnetically induced transparency, lasing without inversion, and ultra-slow group velocity of light have been widely studied. This work is focused on coherent control of refractive index and resonant fluorescence in multilevel medium. We suggest two promising schemes for resonant enhancement of the refractive index with eliminated absorption and propose their implementation in transition element doped crystals with excited state absorption and in a cell of Rb atoms at natural abundance. We show how to use one of these schemes for spatial variation of the refractive index via its periodic resonant increase/decrease, remarkably keeping at the same time zero absorption/gain. It opens the way to production of transparent photonic structures (such as distributed Bragg reflectors, holey fibers, or photonic crystals) in a homogeneous resonant atomic media such as dielectrics with homogeneously distributed impurities, atomic, or molecular gases. These optically produced photonic structures could easily be controlled (including switching on/off, changing amplitude and period of modulation) and would be highly selective in frequency, naturally limited by the width of the optical resonance. We also derive the optical fluorescence spectra of a three-level medium driven by two coherent fields at the adjacent transitions in a general case when all three transitions are allowed. We show that coherent driving can efficiently control the distribution of intensities between the fluorescent channels. In particular, the total intensity of fluorescence at the transition which is not driven by the optical fields may essentially exceed the fluorescence intensity at the driven transitions under the condition of two-photon resonance. This counter-intuitive effect is due to depletion of the intermediate state via atomic interference.

Estuary is a complex region due to the mixing of fresh and sea water as well as the mixing of terrestrial and marine dissolved organic matters (DOM), so that the sorption behavior between hydrophobic organic compounds and dissolved organic matters is very complicated to estimate. By applying fluorescence quenching method, we investigated the influence of the mixing of binary dissolved organic matters from different sources on the binding constant (Kdoc) of pyrene and DOM. Results show that the kinetic of mixing of binary DOMs influences the binding constant, that means if there weren¡¦t enough standing time for mixing of binary DOMs, it would cause some bias in estimating the binding constants of pyrene and DOMs mixture. In this study, we also found that there exists a linear relationship on the binding constants between pyrene and DOMs mixture against mixing ratios. In addition, the mixing mechanism of SRNOM and LHA was dominated by diffusion process from the results of initial mixing efficiency. However, the diffusion process might not be the only mechanism of the mixing reaction of SHHA and LHA, other factors should be considered. Further investigations should be done to find out the dominating factors on the mixing of SHHA and LHA system in the future.

Recently, lifetime imaging has become a subject of intensive research. Lifetime is an important parameter to understand the dynamics of targeted objects and its applications ranges from fluorescence decay of biological objects to relaxation of semiconductor materials and devices. Many methods, such as time-correlated single photon counting (TCSPC) and phase detection in frequency domain, were developed to measure the characteristic lifetime. These methods are now rather matured and widely applied in various studies. However, these methods are only effective for lifetime longer than 100 picoseconds due to the bandwidth limitation of high-speed electronics. For even faster temporal resolution, novel techniques that do not rely on high-speed electronics will be required. In this study, we are integrating an autocorrelator with a galvo-based laser scanning microscope to enable imaging with very high temporal resolution. The principle and technique of pump-probe is implemented through the autocorrelator. In this way, imaging based pump-probe measurements can be realized. Specifically, we have applied the experimental setup so developed in measuring fluorescent dyes and semiconductor devices.

In this dissertation, we construct a temperature controlled sample stage that is compatible with high numerical aperture objective optical microscope, and perform single molecule experiments under the system. Mixing dilute fluorophore (CdSe/ZnS quantum dot, DiI, Rhodamine B) into the liquid crystal matrix (5CB), we monitor the fluorescence dynamics of the individual fluorophore at various temperature. Different from the thermodynamic states of conventional materials, those specific class of materials which we called ¡§liquid crystals¡¨ are attracted for their existence of unique liquid crystal phase, which exhibits a solid-state like higher orientation ordering, and a liquid-state like liquidity. Probe individual fluorophore allows us to monitor the nanometer length scale local structural and dynamic heterogeneity in the solid, liquid crystal and liquid phases. The operating temperature of the platform covers more than 20 oC to 40 oC range with stability much better than 0.1 oC. Quantum dot in PMMA exhibits a clear on-off blinking behavior, and the single exponential fluorescence lifetime relaxation. While in the solid phase of the liquid crystal matrix, quantum dot exhibits similar behavior, which indicates the quantum dot is confined in the matrix. However, there exists slightly difference in decay lifetime. On the contrary, in the liquid crystalline phase as well as the liquid phase, quantum dot exhibits bi-exponential relaxation behavior. Besides a similar time scale relaxation dynamics, there exists additional fast decay behavior, which is from the feasible rotational rotation in the non-rigid matrix. In particular, the anisotropic decay dynamics in the liquid crystalline phase indicates the orientation preference of the liquid crystal molecules. Fluorescence Correlation Spectroscopy (FCS) provides the information of local dynamics of various time scales. FCS results exhibit an unclear transition that crossovers several decades in time scale, which indicates the highly heterogeneity of the liquid crystal. The results of DiI exhibits different rising time in the fluorescence lifetime measurement, which implies the forming of aggregation due to the limited solubility of the DiI molecules in the liquid crystal matrix. Results of Rhodamine B exhibit a clear rotational diffusion dynamics at ~ microsecond scale and the corresponding translational diffusion dynamics at ~ mini-second scale. Moreover, the transition time scale of translational diffusion exhibits a temperature dependence. At higher temperature, it shifts to a shorter time scale.

In this study, we measured the binding constant, KCOC, between several humic acids and benzo(h)quinoline, a nitrogen contained PAHs via using fluorescence quenching method. KCOC of humic acids and phenanthrene, a parent PAHs, is also studied in comparison. Moreover, pH and ionic strength effect on the KCOC were investigated. According to our results, the phenanthrene¡¦s KCOC decreases as the pH increases due to the lower hydrophobicity of humic acid in higher pH values. The variation of benzo(h)quinoline¡¦s KCOC with pH exhibits a more complicated trend, with a maximum value at pH close to the pKb of benzo(h)quinoline. For pH lower than pKb, benzo(h)quinoline is protonated to be benzo(h)quinolinium, a cation, so that the ionic exchange is the dominant prosess in sorption mechanism. Therefore, the binding sites of humic acid increase with pH such that the KCOC increases with pH. In contrast, different mechanism involved in the binding for pH higher than pKb, neutral benzo(h)qunoline becomes dominant and hydrophobic interaction controls the binding prosess in sorption mechanism. At last, the composition of different functional groups of humic acid is also found significant in the binding affinity of benzo(h)qunoline or phenanthrene. Moreover, the benzo(h)qunoline¡¦s KCOC exhibits decreasing trend with increasing magnesium ionic strength because of the reduction of molecular size as well as the benzo(h)qunoline binding sites of humic acid. Findings from this study could provide valuable information for numerical simulation of transport and fates of HOPs in aquatic environment.

The photoabsorption cross sections of NO have been measured in the wavelength range 150~200nm, and the cross sections deviation from Beer-Lambert law due to the insufficient resolution of light source is discussed qualitatively. From the steady state approximation of quenching reactions, the pressure dependence of foreign gas on the ratio of absorption intensity to the fluorescence intensity is derived. Furthermore, the non-Stern-Volmer behaviors of the C2£S(n=0) state of NO were discussed by assuming the non-predissociated (J<9/2) and the predissociated (J>9/2) rotational levels following different quenching mechanisms.

Glycerol kinase (GK) from Escherichia coli is allosterically controlled by fructose 1,6-bisphosphate (FBP) and the glucose-specific phosphocarrier protein IIAGlc of the phosphotransferase system. These controls allow glucose to regulate glycerol utilization. Fluorescence spectroscopic and enzyme kinetic methods are applied to investigate these allosteric controls in this study. The linkage between FBP binding and GK tetramer assembly is solved by observation of homo-fluorescence energy transfer of the fluorophore Oregon Green (OG) attached specifically to an engineered surface cysteine in GK. FBP binds to tetramer GK with an affinity 4000-fold higher than to dimeric GK. A region named the coupling locus that plays essential roles in the allosteric signal transmission from the IIAGlc binding site to the active site was identified in GK. The relationship between the coupling locus sequence in Escherichia coli or Haemophilus influenzae GK variants and the local flexibility of the IIAGlc binding site is established by fluorescence anisotropy determinations of the OG attached to the engineered surface cysteine in each variant. The local flexibility of the IIAGlc binding site is influenced by the coupling locus sequence, and in turn affects the binding affinity for IIAGlc. Furthermore, the local dynamics of each residue in the IIAGlc binding site of GK is studied systematically by the fluorescence anisotropy measurements of OG individually attached to each position of the IIAGlc binding site. The fluorescence steady-state anisotropy measurement provides a valid estimation of the local flexibility and correlates well with the crystallographic B-factors. Steady-state kinetics of FBP inhibition shows that the data are best described by a model in which the partial inhibition and FBP binding stoichiometry are taken into account. Kinetic viscosity effects show that the product-release step is not the purely rate-limiting step in the GK-catalyzed reaction. Viscosity effects on FBP inhibition are also discussed.

Fluorescence resonance energy transfer (FRET)-based biosensors have been designed to fluorometrically detect everything from proteolytic activity to receptor-ligand interactions and structural changes in proteins. While a wide variety of fluorophores have demonstrated effectiveness in FRET probes, several potential sensor components are particularly notable. Semiconductor quantum dots (QDs) are attractive FRET donors because they are rather bright, exhibit high quantum yields, and their nanoparticulate structure enables the attachment of multiple acceptor molecules. Fluorescent proteins (FPs) are also of particular interest for fluorescent biosensors because design elements necessary for signal transduction, probe assembly, and device delivery and localization for intracellular applications can all be genetically incorporated into the FP polypeptide. The studies described in this thesis elucidate the important parameters for concerted QD-FP FRET probe design. Experimental results clarify issues of FRET pair selection, probe assembly, and donor-acceptor distance for the multivalent systems. Various analysis approaches are compared and guidelines asserted based on the results. To demonstrate the effectiveness of the QD-FP FRET probe platform, a ratiometric pH sensor is presented. The sensor, which uses the intrinsic pH-sensitivity of the FP mOrange to modulate the FP/QD emission ratio, exhibits a 20-fold change in its ratiometric measurement over a physiologically interesting pH range, making it a prime candidate for intracellular imaging applications.

The laser-induced fluorescence (LIF) technique is used to visualize and quantify the concentration field around a conductivity probe. The LIF data are compared to the signal collected by the conductivity probe. The objective is to compare the signal of the contact-sensor to the "ground-truth" measurement of the LIF data. Detailed comparison of the temporal response and the peak detection are presented. In addition, a proof-of-concept of a two-color LIF technique is presented using Rhodamine 6G paired with an Argon-ion laser and simultaneously Oxazine 725 paired with a Krypton-ion laser. Optical filters on two digital cameras isolate the emitted light from these respective laser/dye combinations. The objective is to provide detailed quantitation of two over-lapping (non-reactive)chemical plumes.

Thèse doctorat : Images et Systèmes : Villeurbanne, INSA : 2007. / Titre provenant de l'écran-titre. Bibliogr. p. 167-175.