Generation of reproducible attosecond (10-18s) pulses is an exciting goal: in the same way as femtosecond pulses were used to make "movies" of the atomic motion in molecules, attosecond pulses could "uncover" the motion of electrons around nuclei. In this dissertation, we have suggested new ideas that will allow improving one scheme for obtaining such ultra-short pulses: the molecular modulation technique. In a theoretical proposal called Raman Additive technique, we have suggested a method that will allow (with a proper phase stabilization of generated sidebands) to obtain reproducible waveforms of arbitrary shape. An exciting range of possibilities could open up - not only for absolute phase control or sub-cycle shape control, but also for investigation of multiphoton ionization rates as a function of the sub-cycle shape. We have elaborated on the latter subject in another theoretical project, where we have exploited the unique feature of such ultrashort laser pulses, which is synchronization with molecular motion (rotational or vibrational), in order to investigate photoionization of molecules. From experimental point of view, a different construction of driving lasers than previously employed led to establishment of larger molecular coherences at higher operating pressure than in previous experiments. This resulted in simultaneous generation of rotational and vibrational sidebands with only two fields applied. In another experimental proposal using rotational transition in deuterium we have shown that employing a hollow waveguide instead of normal Raman cell improves the efficiency of the generation process. By optimizing gas pressure and waveguide geometry to compensate the dispersion, the method can be extended to efficiently generate Raman sidebands at a much lower energy of driving fields than previously employed. At the end, a very exciting possibility for controlling the molecular motion in a Raman driven system will be shown. Based on the interference effects (EITlike) that take place inside of a molecule, selectivity of different degrees of freedom can be achieved (for example switching from rotational-vibrational motion to pure rotational).

Amplificadores Ãticos amplificam a luz incidente atravÃs de emissÃo estimulada, o mesmo mecanismo que à usado pelos lasers. Com certeza, um amplificador Ãtico, nÃo à nada mais do que um laser sem realimentaÃÃo. Seu principal ingrediente à o ganho Ãtico que à percebido quando o amplificador à sujeito a um bombeio (oticamente ou eletricamente) para conseguir a inversÃo de populaÃÃo nos subniveis. O ganho Ãtico, em geral, depende nÃo somente da freqÃÃncia (ou comprimento de onda) do sinal incidente, mas tambÃm da intensidade do feixe local em qualquer ponto dentro do amplificador. Esse trabalho foi motivado por uma procura contÃnua do conhecimento e entendimento das caracterÃsticas e dos fenÃmenos envolvidos na amplificaÃÃo de regime de pulso curto que seriam relevantes como aplicaÃÃes para processos nos quais tais fenÃmenos nÃo podem ser negligenciados. Sem perda de generalidade, evitamos sistemas de vÃrios canais, consideramos aqui um Ãnico canal com relaÃÃo a outro, pelo fato de que o ganho e o Ãndice de refraÃÃo ambos dependem do nÃmero de canais envolvidos. Neste trabalho foi simulada inicialmente a amplificaÃÃo Ãptica onde o ganho era constante de modo a comparar com um novo modelo proposto aqui, aonde o ganho à periÃdico. Neste caso modelamos as parcelas de transferÃncia de energia do bombeio e do sinal em funÃÃes periÃdicas de onde foi simulado com diferentes parÃmetros das funÃÃes periÃdicas escolhidas. AlÃm do mais, os efeitos de dispersÃo, automodulaÃÃo de fase, pulso walkoff, efeito Raman e depleÃÃo de pulso foram considerados como fatores importantes para amplificaÃÃo Raman de pulsos curtos. Com relaÃÃo à forma dos pulsos de bombeio e a semente Raman para as simulaÃÃes toma um pulso Gaussiano e um sinal CW fraco respectivamente. O pulso de bombeio transfere energia para o sinal CW ao longo da fibra. Todas as simulaÃÃes foram realizadas usando um mÃtodo numÃrico espectral bem conhecido como Split-Step Fourier Method resolvendo as equaÃÃes acopladas nÃo lineares de SchrÃdinger. / Optical Amplifiers amplify incident light through stimulated emission, the same mechanism which is used by lasers. Indeed, an optical amplifier, it is not but a laser without feedback. Its main ingredient is optical gain which is realized when the amplifier is under pumping process (optically or electrically) in order to cause population inversion at electronic sublevels. In a long run, the optical gain will not only depend on frequency (wavelength) of incident signal, but it also depends on the local beam intensity of the optical gain that is entailed to the amplifier medium. This thesis was stimulated by the continuous pursue of knowledge and understanding of characteristics and phenomena involved in the Raman amplification process in the regime of short pulses which would be relevant as the appliance for processes in which such phenomena can not be neglected. Without loss of generality, we considered the case of where there is an only one channel to another one by the fact that the gain and the refractive index both depend on the number of channels. In this thesis, it has also been simulated the optical amplification where the gain was constant in order to comparing to the periodic gain presented in this thesis. In addition, the effects of dispersion, self phase modulation, pulse walk-off, Raman effects and pulse depletion were considered as important factors for Raman amplification of short pulses. That was also considered for our simulations a weak CW signal or a Raman seed to be amplified by an intense pump Gaussian pulse. All the simulations were achieved using a well-known spectral numerical method namely Split-Step Fourier Method for solving the coupled Nonlinear SchrÃdinger Equations.

Abstract The first hyperpolarizability(£]) of five charge-transfer molecules are determined using the hyper-Rayleigh scattering (HRS) technique at two excitation wavelengths : 1064nm and 1907nm. The 1064nm excitation wavelength is derived from a Nd: YAG pulsed laser, and the 1907nm excitation wavelength is obtained by shifting the 1064nm laser light by stimulated Raman scattering of pressurized H2 gas. Four of the five samples contains thiophene and thiazole are synthetized by Prof. Shu Ching-Fong at the National Chiao Tung University (NCTU) and the other sample is synthetized by Prof. Hong, Jin-Long at the National Sun Yat-sen University (NSYSU). The measured £] values are used to calculate the intrinsic molecular hyperpolarizabilities using the two-level model previously developed by Oudar and Chemla. The results are related to molecular structure.

This work reports on the construction of a 408 nm laser system designed to drive stimulated Raman transitions between the F = 4 and F = 5 2 S 1/2 states of 87 Sr + using the 2 P 3/2 state as the intermediate state. This laser system will be used as part of a 87 Sr + ion interferometer. This work also includes a discussion of relevant theory describing the interaction of the ions and laser, along with a calculation of the transition rates as a function of laser power and detuning.

This thesis presents the experimental work performed on the spectrin protein. The aim of the work was to study the direct interactions of spectrin, the cytoskeleton of RBCs, with membrane lipid to determine its effects on the mechanical properties of the lipid bilayer. Motivation for this work came from a lack of unanimity in the field of spectrin, and the hypothesized potential of the protein to perforate giant unilamellar vesicles. The work aimed to investigate and determine how spectrin-lipid interactions influence membrane mesoscopic morphology and biophysics in ways that could ultimately be important to cellular function. For this purpose, a protocol was implemented to take into account the different aspects of the binding. Direct visualisation of the spectrin-lipid interaction and distribution was achieved using confocal fluorescence microscopy. Changes in the mechanical properties of the membrane were investigated using the micropipette aspiration technique. Finally the thermodynamics of the interaction were considered with isothermal titration calorimetry experiments. This allowed evaluation of the protein-lipid interaction in a complete and coherent manner. Experiments were also performed on another elastic protein, alpha-elastin, for comparison. In addition to its similarities with spectrin (both possess hydrophobic domains and entropy elasticity), elastin is auto-fluorescent which makes it an attractive model protein. Elastin was also used as a sample model to implement new techniques using nonlinear optics microscopy.

Cette thèse porte sur les mélanges d’onde non linéaires qui peuvent avoir lieu dans le champ évanescent de nanofibres de silice. Nous nous sommes plus particulièrement intéressés à la diffusion Raman stimulée qui est obtenue par l’interaction du champ évanescent très intense et un liquide non linéaire dans lequel baigne la nanofibre. Afin de mettre en évidence la diffusion Raman stimulée« évanescente », nous avons développé un modèle de simulation non linéaire dont le but est de déterminer les caractéristiques des nanofibres à réaliser. Le gain Raman modal est calculé afin de trouver le rayon optimal des nanofibres pour chaque liquide ou mélange de liquides possible. En considérant la puissance critique et le seuil de dommage de nos nanofibres, nous avons déduit la longueur minimale d’interaction. Les conditions d’adiabacité des parties évasées menant à la nanofibre sont également discutées. Ces spécifications nous ont amenés à développer une plateforme de tirage de nanofibres spécifiquement dédiée à ces expériences de non-linéarités évanescentes. Cette palteforme nous permet de tirer des nanofibres de diamètre allant jusqu’à 200 nm sur des longueurs de 10 cm, avec plus de 90% de transmission. Avec ces nanofibres, nous avons mis en évidence le premier ordre Stokes de l’éthanol dans le champ évanescent d’une nanofibre, ainsi que les premier et second ordres Stokes du toluène. Ces premières expériences sont en très bon accord avec nos simulations et ouvrent la voie à de nombreuses expériences en optique non linéaire. / The present PhD thesis explored nonlinear wave mixing with the strong evanescent field of nanofibers. The focus has been on the effect of stimulated Raman scattering which is activated by the interaction between such a strong evanescent field and the nonlinear liquid surrounding the nanofiber. In order to observe the stimulated Raman scattering, we investigated the nonlinear modeling to determine the needed characteristics of the nanofibers. The modal Raman gain was calculated to determine the optimal radius of nanofibers for each possible liquid. Considering the critical power and the damage threshold of our nanofibers, we found the minimum required interaction length. The condition of adiabacity of the tapers was also described. These specifications of nanofibers guide us towards the design of a proper pulling system. Several pulling systems and techniques are investigated for the fabrication of our specific nanofibers. We now are able to fabricate low loss uniform nanofibers of up to 10 cm long, a diameter down to 200 nm, with two identical low loss tapers by using our own designed translation stage pulling platform and implemented with the “variable heat brush” technique. With the achieved nanofibers, the Raman effect induced in the evanescent field was observed in both pure (ethanol) and binary mixture (toluene in ethanol) liquids. These first measurements are in good agreement with our simulation even without any fitting parameters in the modeling.

[pt] Este trabalho visa estabelecer na prática a estatística do ganho dependente da polarização (PDG-Polarization Dependent Gain) em sistemas ópticos com amplificação Raman. A amplificação Raman depende fortemente da polarização relativa entre os fótons de bombeio e de sinal, que tem que ser paralelas para máximo ganho [1]. Portanto, a birrefringência é um importante limitador de desempenho de sistemas de longa distância amplificados por esta técnica visto que modifica os estados de polarização de forma diferente para cada comprimento de onda. A birrefringência varia aleatoriamente de acordo com as flutuações do ambiente onde está a fibra óptica, dando origem à dispersão dos modos de polarização, efeito este conhecido pela sigla PMD. Alguns experimentos recentes mostraram que o amplificador Raman não só depende do estado de polarização do sinal de entrada, mas também que o valor da dependência do ganho com a polarização (PDG- Polarization Dependent Gain) flutua devido a natureza aleatória da PMD [4,5]. É importante conhecer a estatística da PDG, sua relação com a PMD e como a PDG pode ser reduzida a níveis aceitáveis. Nesse trabalho será medida experimentalmente a distribuição estatística da PDG em fibras de dispersão deslocada e os resultados comparados com as previsões teóricas dadas por [2]. / [en] Raman amplifiers are very attractive because they provide a large and relatively flat gain over a wide bandwidth while maintaining a small noise figure, and they can be made using regular silica fiber. However, the Raman Gain coefficient is polarization sensitive and can be up to ten times higher when the signal and pump polarization states are parallel rather than perpendicular [1]. Usually fibers present some degree of residual asymmetry - because the fiber core is slightly out-of-round, or because of mechanical stress on the deployed fiber - and this causes polarization mode dispersion. The light traveling along one polarization axis moves slower or faster than the light polarized along the other axis. This effect distorts the signal and causes polarization fluctuations along the fiber. As the Raman gain is higher when the signal and pump polarization states are parallel these fluctuations of the relative polarization between signal and pump vary the instantaneous value of the Raman gain. So the Polarization Dependence Gain (PMG) is directly related to the PMD. Several experimental studies have shown not only that the gain of raman amplifiers depends on the state of polarization of the input signal but also that this polarization-dependent gain (PDG) fluctuates over a wide range because of the random nature of polarization mode dispersion (PMD) [4,5]. It is important to know the statistics of PDG, its relationship to the PMD, and how the PDG can be reduced to acceptable low levels. In this letter we will demonstrate experimentally the statistical distribution of the PDG given by [2]. This In this work the polarization dependent gain (PDG) fluctuations due to PMD in Raman amplified optical transmissions is experimentally demonstrated.

DNA is packaged into the nucleus of a mammalian cell as a nucleoprotein complex called chromatin. Changes in chromatin structure occur during processes that are critical to an understanding of mammalian cell biology such as cell division. Existing fixed-cell techniques have provided insight into chromatin organization in the mammalian nucleus. In addition, fluorescence microscopy techniques have allowed us to study changes in chromatin structure in living cells. However, most of these fluorescence techniques cannot be used for tissue imaging or long-term imaging due to photobleaching. In this thesis, we demonstrate that a label-free technique called Stimulated Raman Scattering (SRS) microscopy can be used to solve these problems and study chromatin structure in living mammalian cells both in culture and in tissue. SRS is a vibrational microscopy technique that takes advantage of intrinsic contrast arising from specific chemical bonds in a molecule. Nucleic acids have specifc phosphate and CH vibrations that can be used to determine their cellular distributions. Imaging at specific phosphate peaks using fingerprint SRS microscopy allows the detection of polytene chromosomes in Drosophila salivary gland cells and condensed chromatin in metaphase mammalian cells. In addition, we develop a technique called multicolor SRS microscopy, in which we image at several wavelengths across the CH vibrational band, and then use linear combination to simultaneously determine the nucleic acid, lipid and protein distributions in living mammalian cells. This technique achieves greater contrast than imaging at the phosphate vibrational peak due to the stronger SRS signal in the high wavenumber CH band and so allows us to determine chromatin structure in interphase mammalian cells. This technique also allows long-term imaging of living mammalian cells and the imaging of tissue such as mouse skin. The technique is used to monitor mammalian cell division in culture and paves the way for similar studies in living tissue. This technique will provide insight into cell division, differentiation and apoptosis during development and in disease models such as cancer.

The incidence of gastrointestinal (GI) cancers has been steadily increasing in the UK since the mid 1970’s. These include cancers of the colon and oesophagus. Colon cancers have a high incidence rate, being the fourth most common cancer in the UK for both men and women. Oesophageal cancers in comparison are much rarer, however they have a poor survival rate primarily due to a late diagnosis. The key to improving survival for these cancers and many others is to detect and remove the disease at the early stages, to prevent the cancer from advancing. At present the ‘gold standard’ for diagnosis is a biopsy followed by histopathology. This technique is invasive, time consuming and highly subjective. It is therefore important to look towards non-invasive methods for early and rapid diagnosis. Optical techniques have begun to show such promise. By probing the interactions of tissues with light, diagnostic information is able to be obtained non-invasively. Techniques such as Raman spectroscopy utilise inherent molecular vibrations to extract biochemical information from tissues. Raman spectroscopy, however, is currently fundamentally limited by long acquisition times, due to the inherently weak signals produced. Using coherent Raman techniques such as coherent anti-Stokes Raman scattering (CARS) and stimulated Raman scattering (SRS), the molecular vibrations are coherently driven to provide an enhancement in signal. This thesis explored spectral signatures from snap frozen oesophageal sections in the fingerprint (450 cm-1 to 1850 cm-1) and high wavenumber (2800 cm-1 to 3050 cm-1) regions using spontaneous Raman and compared with spectra from hyperspectral SRS. The diagnostic potential for each technique was assessed for four major pathology groups, normal, Barrett’s oesophagus, dysplasia and adenocarcinoma. Samples were classified using a principal component fed linear discriminant analysis (PCA-LDA) approach with a leave-one-out cross validation. Comparisons were made to haematoxylin and eosin (H&E) stained sections. Raman in the fingerprint region was found to be the most promising for diagnosis. There were minimal changes in the high wavenumber region between pathology groups which was also reflected in the SRS spectra and proved to be insufficient for classification. Further comparisons were made between spontaneous and coherent Raman techniques using frozen colon sections. The morphological and structural information available was explored using a k-means cluster analysis. Both spontaneous and coherent Raman were able to distinguish important structural features in the colon, such as the epithelial cells that form the colonic glands and surrounding connective tissue. Both are important visual markers for cancer diagnosis in the current approach. SRS demonstrated higher spatial resolution and faster acquisition times in comparison to spontaneous Raman. This work has discussed the many advantages of using coherent Raman techniques for tissue applications, but has also highlighted some of the limitations for spectral measurements, arising from the complexity of the system.

Volumetric optical microscopy has the advantages of quantitative and global measurement of three-dimensional (3D) biological specimens with high spatial resolution and minimum invasion. However, current volumetric imaging technologies based on light transmission, scattering or fluorescence cannot reveal specimen’s chemical distribution that brings insights to study the chemical events in organisms and their metabolism, functionality, and development. Stimulated Raman scattering (SRS) microscopy allowing visualization of chemical contents based on their intrinsic molecular vibrations is an emerging imaging technology to provide rapid label-free volumetric chemical imaging. This dissertation describes three methodologies for developing advanced volumetric SRS imaging technologies to address the challenges of imaging in vivo samples, imaging speed, and axial resolution. In the first methodology, SRS volumetric imaging is enabled by axially scanning the laser foci for sectioning different depth layers. In Chapter 2, we utilize a piezo objective positioner to drive the objective. Combining with the tissue clearance technique, we realize volumetric SRS imaging up to 500 µm depth in brain tissues showing the potential for 3D staining-free histology. The limitations of piezo scanning are slow speed and disturbance to in vivo samples while rapidly scanning the objective. To tackle the limitations, in Chapter 3, we develop a remote-focusing volumetric SRS microscope based on a deformable mirror and adaptive optics optimization, allowing focal scanning without physically moving the objective or sample. We demonstrate in vivo monitoring of chemical penetration in human sweat pores. In the second methodology, instead of axially scanning the laser foci, the SRS volumetric imaging is enabled by projection imaging with extended depth-of-focus (DOF) beams such as Bessel beams and low numerical-aperture beams. The extended DOF beams integrate SRS signals along the propagation direction to form projection images; thus, a single lateral scan obtains the volumetric chemical information, significantly increasing the volumetric imaging speed for measuring chemical content over a large volume. In Chapter 4, we describe a stimulated Raman projection microscope for fast quantitation of chemicals in a 3D volume. However, projection imaging intrinsically loses axial resolution. We addressed the limitation by developing SRS projection tomography. Mimicking computed tomography, the axial information is reconstructed by angle-dependent projection images obtained by sequentially rotating the sample in a capillary glass tube within the SRS focus. Nevertheless, sample rotation is complicated and not compatible with in vivo samples. To address the difficulty, in Chapter 5, we develop tilted-angle-illuminated stimulated Raman projection tomography which utilizes tilted-angle beams with a tilted angle respected to the optical axis of the objective to obtain angle-dependent projections. This scheme is free of sample rotation and enables fast projection scanning for pushing the imaging speed. The calibration approach and vector-field back-projection algorithm are developed for the multi-view tomographic reconstruction. In the third methodology, we improve the spatial resolution in miniature volumetric SRS imaging via the innovation of metasurface photonics. In developing an SRS endoscope for volumetric chemical imaging inside the human body, the axial resolution deteriorates due to chromatic and monochromatic aberrations induced by poorly made miniature objective lenses. In Chapter 6, we develop a silicon metasurface tailored for compensating the phase errors between the pump and Stokes wavelengths of a singlet refractive lens. Integrating the metasurface with the refractive lens, the hybrid achromatic metalens is compact and provides nearly diffraction-limit resolution, demonstrating a way for developing high resolution chemical imaging endoscopy.