<p> 3D printing has established itself as a critical tool for manufacturing in all areas. It has evolved from a purely rapid prototyping technique into a feasible process for large-scale processing. A wide variety of 3D printing processes exist across an extreme range of size, from meters to nanometers. Much of the current technological advances come from pushing fabrication techniques to smaller and smaller scales. For 3D printing this has led to the rise of two-photon polymerization, a direct laser writing process with submicron structuring capabilities. Two-photon polymerization has proven its worth as a nanoscale 3D fabrication technique but is often considered slow and expensive, two undesirable qualities for high throughput manufacturing. Parallelization methods such as projection lithography are potential solutions to increasing the throughput capabilities of two-photon polymerization 3D printing. Additionally, the drive for further reducing the print size has inspired printing resolution enhancing strategies in two-photon polymerization printing by processes such as stimulated emission depletion (STED) and other STED-inspired pathways. This work will explore avenues for improving two-photon polymerization printing throughput and resolution.</p> <p> First, a two-photon polymerization printing system is constructed with a secondary laser for controlling polymerization inhibition. Through a STED process, a 65 nm wide printed line feature was achieved. Alongside this, a characterization and verification methodology for choosing new photoinitiator molecules for similar inhibition lithography processes is presented. Through implementation of tests such as Z-scan, fluorescence depletion, ultrafast transient spectroscopy and UV-Vis absorption and fluorescence measurements a promising new photoinitiator with 5-factor improvement in printing efficiency is found. </p> <p> Second, a projection lithography scheme is developed for rapid two-photon 3D printing. A digital micro-mirror device (DMD) is utilized for dynamic pattern generation and the effects of its dispersion properties are considered. Through a spatiotemporal focusing process, continuous 3D printing is achieved at vertical prints speeds of 1 mm s-1. Simulations performed representing this rapid printing process indicate a ~1 µm layer print feature size for large areas of exposure. Comparably, a printed vertical feature size of ~ 1 µm was achieved. Lateral feature sizes ~200 nm were also demonstrated in fabrication. A variety of complex 3D structures are printed for demonstration of the spatiotemporal focusing projection lithography process including millimeter scale objects with micrometer scale 3D features.</p> <p> Finally, resolution enhancing strategies are implemented into the continuous, projection two-photon lithography technique. An investigation of the inhibition properties of a variety of photoinitiator systems for inhibiting polymerization achieved with low repetition rate laser exposure is presented. A planar polymerization inhibiting region is generated by creating a light sheet propagating perpendicularly to the projection printing plane. </p>

In this thesis, advanced spectroscopy methods are discussed and applied to gain understanding of the physical properties of organic conjugated molecules, II-VI thin film semiconductors, and vertical cavity surface emitting lasers (VCSEL). Experiments include single photon and two-photon excitation with lasers, with subsequent measurements of the absorption and photoluminescence, as well as photocurrent measurements using tungsten and xenon lamps, measuring the direct current and the alternating current of the devices. The materials are investigated in dissolved form (conjugated polymers), thin films (polymers, II-VI semiconductors), and complex layer structures (hybrid device, VCSEL). The experiments are analyzed and interpreted by newly developed or applied theories for two-photon saturation processes in semiconductors, bandgap shrinkage due to optically induced electron hole pairs, and the principle of detailed balance to describe the photoluminescence in thin film cadmium sulfide. / Ph. D.

Nanoscale materials have great applications in many areas. One of these applications is for manufacturing ultra-compact and efficient sensors for chemical and biological molecule detection. Noble metals, such as gold (Au) and silver (Ag), because of their distinguished optical property"localized surface plasmon resonances (LSPRs) that exhibit low loss, are ideal materials to fabricate these nanoscale plasmonic particles or structures. This work addresses the synthesis, characterization, and sensing applications of Au and Ag nanoparticles (NPs). The progress on certain subjects related to our work"NP synthesis, surface functionalization, Au sphere-film structure and two-photon fluorescence"are reviewed in Chapter 1. We also show the calculation results of LSPRs of Au nanosphere suspensions using Mie theory. The measured extinction spectra of Au nanosphere suspensions agree with the calculated results very well. Chapter 2 is a chapter describing the chemical synthesis of a variety of NPs, such as Ag prisms and cubes, Au spheres, rods, and bipyramids. These experiments involved different synthetic mechanisms and methods which enabled us to prepare NPs with desired shapes and optical properties. To put these NPs into application, it is desirable and sometimes necessary to functionalize their surfaces. In Chapter 3, we present the functionalization of Ag cubes with poly(allylamine hydrochloride) (PAH) and poly(allylamine hydrochloride)-dithiocarbamate (PAH-DTC), which follows our previous work on Au NPs. The purpose of studying Ag instead of Au is to use the stronger plasmonic enhancement in Ag when applied to two-photon imaging applications. However, we found that PAH-DTC shrank the Ag cubes. We also functionalized the cationic hexadecyltrimethylammonium bromide (CTAB)-stabilized Au NRs with anionic poly(sodium 4-styrenesulfonate) (PSS). Coated with the strong polyelectrolyte PSS, the NRs become more manageable and can be stable for over six months and are easily immobilized onto positively charged substrate. We put PSS-functionalized Au NPs into use and studied their adsorption process onto PAH-coated optical fiber tapers by monitoring the transmission light through the fiber. When the diameter of the fiber taper gets smaller, stronger coupling occurred between transmitted light inside the taper and the Au NPs on the taper surface (cylinder). This coupling resulted in a loss of the guided light at the plasmon resonance wavelength of the NPs. By monitoring this loss, we can study the adsorption rate of Au NPs onto the fiber. In Chapter 4, we used Au nanospheres to study the adsorption rate on substrates with different curvatures. We also established a theoretical model to explain this phenomenon for cylindrical surface as well as planar and spherical surfaces. Our results fit well with the theory, which predicts that particle adsorption rates depend strongly on surface geometry, and can exceed the planar surface deposition rate by over two orders of magnitude when the diffusion length of the particle is large compared to the surface curvature. In Chapter 5, we studied the optical properties of Au nanospheres separated from a thick Au film by a polyelectrolyte multilayer (PEM) film assembled from PAH and PSS under specific pH condition. The PEM film undergoes swelling and shrinking when the environmental pH is changed as a result of charging and discharging of the polyelectrolytes. Therefore, the PEM film provides an efficient means to tune the distance between Au spheres and Au film. The extinction peak blue-shifted as much as 100 nm when the pH of the water changed from pH 10 to pH 3 for 100 nm diameter Au spheres on a PEM film assembled at pH 9.5. Our preliminary estimates that the gap between sphere and surface can be as small as a few nm even though the film itself is tens of nm thick when it is not constrained by Au spheres. We studied two-photon excitation fluorescence (TPEF) from Ag triangles in Chapter 6. The triangles were fabricated by nanosphere lithography, which used convective self-assembly to make the nanosphere mask. The LSPRs of the nanotriangles were tuned to be in the 800--900 nm range to match with the Ti:Sapphire pulse laser at 880 nm. We found that certain spots on the fluorescence images gave rise to larger fluorescence intensity than rest of the area. SEM imaging reveals that the unusually bright spots seen on the surface were related to regions where the triangles transformed to spherical particles. The larger intensity is tentatively ascribed to the plasmon resonance of those spherical particles in ~400 nm range. / Ph. D.

As the boundaries of Moore's Law rapidly approach, research is increasingly turning to exotic materials and metamaterials to advance the capabilities of electronic and micromechanical systems. This thesis presents systems, models, and simulation techniques for novel microfabrication in the realms of silicon carbide (SiC) and metal mesh filters. Through the unification of femtosecond laser pulses, high numerical aperture (NA) objective lenses, and system power and motion control, a system capable of achieving arbitrary 3D features without line-of-sight with aspect ratios up to 109:1 in SiC is developed. Additionally, a model of a direct-write femtosecond-laser-ablation fabricated metal mesh filter (MMF) is simulated in ANSYS High Frequency Structure Simulator (HFSS) and validates the novel fabrication technique. Finally, a transmission-line-theory based MMF model is presented as an alternative to modelling multilayer filters in HFSS. This model produces comparable results to an HFSS simulation with dramatically reduced computational intensity.

La fluorescence excitée à deux photons est actuellement largement utilisée pour l’imagerie de tissus biologiques, mais la faible sensibilité des fluorophores utilisés en microscopie confocale (excitation à un photon) à une excitation à deux photons (ADP) rend nécessaire la conception et la synthèse de nouveaux fluorophores spécifiques pour la microscopie de fluorescence par excitation bi-photonique (MFEB). Mon travail de thèse a ainsi porté sur l’ingénierie moléculaire (conception, synthèse et caractérisations) de nouveaux fluorophores pour la MFEB. Nous nous sommes particulièrement intéressés à des systèmes unidimensionnels (1D) de petite taille comportant des systèmes π étendus autour d’un cœur rigide (dicétopyrrolopyrrole ou DPP) et entourés de différents systèmes électro-actifs. Nous avons modifié par la suite les fluorophores précédents de manière à pouvoir les conjuguer à des molécules d’intérêt biologique, comme des protéines. Ces fluorophores bio-conjugables ont été greffés sur un peptide du virus HIV étudié au laboratoire : TAT (Trans-Activator of Transcription). L’imagerie par microscopie biphotonique a été effectuée avec succès sur des cellules HeLa. Nous nous sommes ensuite tourné vers la mise au point de nouvelles sondes multimodales pour associer la MEBP à une autre modalité d’imagerie : la résonance magnétique nucléaire et la microscopie électronique (imagerie corrélative). Pour ce faire nous avons développé des colorants fluorescents par excitation bi-photonique comportant une entité paramagnétique ou dense aux électrons (nanoparticules de magnétite, ion gadolinium III ou atomes lourds comme le platine et l’or). / Two-photon induced fluorescence is nowadays widely used for the imaging of biological tissues. The classical fluorophores used in confocal microscopy exhibit low sensitivity to two-photon excitation for the two-photon excitation microscopy (TPEM), led the researchers towards the development of new fluorophores, specifically engineered for TPEM. This manuscript describes our work on conception, synthesis and characterizations of new one-dimensional fluorophores based on dicétopyrrolopyrrole (DPP) central core, surrounded by various electro-active systems through π conjugated systems. We also modified such fluorophores to be able to conjugate them to molecules of biological interest, such as proteins. These bio-conjugable fluorophores were grafted on a peptide of HIV virus studied in our laboratory: TAT (Trans-Activator of transcription). The imaging by TPEM was successfully performed on HeLa cells. In addition we developed new multimodal probes for the correlative light electronic microscopy and for the correlative imaging fluorescence microscopy/ Magnetic resonance imaging (MRI). Theses multimodal probes associate a fluorescent moiety based on the DPP core associated to a paramagnetic or electron dense entity (magnetite nanoparticles, gadolinium III or heavy atoms such as platinum or gold).

La photothérapie dynamique (PTD) est un traitement alternatif du cancer qui nécessite l’utilisation de chromophores (photosensibilisateurs) capables d’induire la mort cellulaire suite à une irradiation lumineuse. Les nanoparticles d’or (AuNP), grâce à leur phénomène de résonance plasmon localisée, peuvent exalter les propriétés photophysiques des chromophores localisés à leur surface. L’utilisation d’une excitation biphotonique, dans le proche infrarouge, peut être utilisée pour améliorer l’action thérapeutique (PTD) ou diagnostique (imagerie de fluorescence) des chromophores en augmentant la profondeur de pénétration dans les tissus et la résolution tridimensionnelle de la microscopie biphotonique.Lors de ce travail, l’élaboration de nouvelles nanoparticules hybrides est proposée, présentant des applications potentielles en bioimagerie (sondes brillantes) et comme photosensibilisateurs pour la PTD. Ces nanoparticules sont composées d’un cœur d’or sur lequel sont greffés des conjugués polymère-chromophores biphotoniques. La stratégie de synthèse a consisté à : i) synthétiser des conjugués polymère-chromophores biphotoniques solubles dans l’eau ; ii) les greffer de manière orientée à la surface des AuNP. La synthèse des conjugués polymère-chromophores hydrosolubles on été synthétisés via le couplage efficace de chromophores hydrophobes en position latérale des copolymères P(NAM-co-NAS) bien-définis, obtenus par la techniques de polymérisation radicalaire contrôlée RAFT. Cette stratégie permet le contrôle à la fois de la longueur des chaînes polymère formées (2 000 g.mol-1 < Mn plus < 37 000 g.mol-1) et du nombre de chromophores couplés par chaîne (de 1 à 21). Le greffage orienté de ces conjugués à la surface des AuNP a été mis en évidence (TEM, ATG) et les densités de greffage se sont avérées élevées (~0.5chaîne/nm²). Un des rôles de la chaîne polymère étant de contrôler la distance entre les chromophores et la surface des AuNP. Il a été mis en évidence que l’augmentation de la longueur de chaine des conjugués induisait, outre une augmentation de la couronne greffée (par MET) sur les AuNP, une augmentation de l’émission de fluorescence de ces conjugués polymère-chromophores. Enfin, les propriétés biologiques des conjugués avant et après greffage sur les AuNP ont été évaluées in cellulo, mettant en valeur leur potentiel pour des applications thérapeutiques et diagnostiques. / Photodynamic therapy (PDT) is an alternative treatment of cancer requiring the use of chromophore molecules (photosensitizers), which can induce cell death after light excitation. Gold nanoparticles (AuNP), exhibiting localized Surface Plasmon Resonance, can enhance the photophysical response of chromophores located in their vicinity, and thus improve their therapeutic action. Moreover, the use of highly localized two-photon chromophores (photosensitizers and fluorophores), capable to undergo a localized excitation by light in the Near InfraRed region, should increase the penetration depth into tissues, thus improve the treatment efficiency (by PDT) and the imaging (by fluorescence microscopy) of cancer tissues.In this work, we describe the elaboration of water-soluble hybrid nano-objects for PDT and fluorescence bioimaging applications, composed of two-photon chromophore-polymer conjugates grafted onto gold nanoparticles. In order to obtain these nano-objects we follow a multistep strategy: i) the synthesis of a well-defined water-soluble chromophore-polymer conjugates; ii) the end-group oriented grafting of chromophore-polymer conjugates onto 20 nm AuNP. The coupling of hydrophobic two-photon chromophores on linear water-soluble copolymer chains (poly(N-acryloylmorpholine-co-N-acryloxysuccinimide)), obtained by controlled/living RAFT polymerization, resulted in well-defined water-soluble chromophore-polymer conjugates, with different polymer lengths (2 000 g.mol-1 < Mn < 37 000 g.mol-1) and architectures (random or block), and a controlled number of chromophores per chain (varying between 1 and 21). Their grafting onto 20 nm AuNP gave water-soluble hybrid nano-objects with high grafting densities (~0.5 chains/nm²). The role of the polymer chain being to tune the distance between chromophores and AuNP surface, we have evidenced the increase in the polymer corona thickness of grafted AuNP (estimated by TEM) with the increasing polymer Mn, corroborating with the corresponding distance-dependent fluorescence properties of those. Finally, the in cellulo biological properties of two-photon chromophore-polymer conjugates, before and after grafting onto AuNP, have been investigated, highlighting their potential for two-photon bioimaging and PDT applications.

The field of organic chemistry is divided into many subfields, which include polymer design and synthesis, transition metal catalysis and organocatalysis among a variety of others. Challenges in polymer design and synthesis can be highlighted pointedly in the use of photoresists for lithographic processing. Recent challenges in development of shorter wavelength sources has led to the need to develop new photoresist materials that can be exposed twice without any development steps in between. Two methods for addressing double exposure materials will be presented. Additionally, the areas of catalysis, whether transition metal or organic in nature, are important methods in organic synthesis. The mechanism of the addition of Gilman reagents to enones has been the subject of debate, and efforts to elucidate this mechanism will be presented. Finally, organocatalysis has expanded its scope into a variety of reactions previously only conducted with transition metal catalysts. Work towards an enantioselective allylic amination reaction using organocatalysis as well as absolute stereochemistry of the product will be explored. / text

Articular cartilage has been imaged using the following multi-photon modalities: Second Harmonic Generation (SHG), Two-photon Fluorescence (TPF) and Coherent Anti-Stokes Raman Scattering (CARS). A simple epi detection microscope was constructed for SHG and TPF imaging in the early stages of this research. Later the imaging was transferred to a new microscope system which allowed simultaneous forwards and epi detection and combined CARS imaging with TPF and SHG. Multiphoton spectroscopic studies were conducted on both intact tissue samples and the major components of the extracellular matrix, in order to identify sources of TPF. Fluorescence was detected from type II collagen, elastin and samples of purified collagen and elastin crosslinks. Age related glycation crosslinks of collagen may be a significant source of TPF. No fluorescence was detected from proteoglycans. In intact, unfixed healthy articular cartilage the cells were observed via CARS, surrounded in their pericellular matrix which is characterised by an increase in TPF. The collagen of the extra cellular matrix showed up clearly in the SHG images. Diseased cartilage was also imaged revealing microscopic lesion at the articular surface in early osteoarthritis and highly fibrous collagen structures and cell clusters in more advanced degeneration. In young healthy cartilage a network of elastin fibres were found lying parallel to the articular surface in the most superficial 50μm of the tissue. Regional variations in these fibres were also investigated. The fibres appeared mainly long and straight suggesting that they may be under tension, further work is needed to identify whether they have a mechanical function. The polarization sensitivity of the SHG from collagen has been investigated for both cartilage and tendon. In the most superficial tissue these measurements can be used directly to determine the collagen fibre orientation. However at increasing depths the effects of biattenuation and birefringence must be considered. Healthy cartilage has a characteristic pattern of polarization sensitivity with depth and this changes at lesions indicating a disruption of the normal collagen architecture. The methods developed in this thesis demonstrate the use of non-linear microscopy to visualise the structure of the extracellular matrix and cells in intact unstained tissue. They should also be appropriate in many areas of cell and matrix biology.

Optical imaging methods have the potential to detect ovarian cancer at an early, curable stage. Optical imaging has the disadvantage that high resolution techniques require access to the tissue of interest, but miniature endoscopes that traverse the natural orifice of the reproductive tract, or access the ovaries and fallopian tubes through a small incision in the vagina wall, can provide a minimally-invasive solution. We have imaged both rodent and human ovaries and fallopian tubes with a variety of endoscope-compatible modalities. The recent development of fiber-coupled femtosecond lasers will enable endoscopic multiphoton microscopy (MPM). We demonstrated two-and three-photon excited fluorescence (2PEF, 3PEF), and second-and third-harmonic generation microscopy (SHG, THG) in human ovarian and fallopian tube tissue. A study was undertaken to understand the mechanisms of contrast in these images. Six patients (normal, cystadenoma, and ovarian adenocarcinoma) provided ovarian and fallopian tube biopsies. The tissue was imaged with three-dimensional optical coherence tomography, multiphoton microscopy, and frozen for histological sectioning. Tissue sections were stained with hematoxylin and eosin, Masson's trichrome, and Sudan black. Approximately 1 mu m resolution images were obtained with an excitation source at 1550 nm. 2PEF signal was absent. SHG signal was mainly from collagen. 3PEF and THG signal came from a variety of sources, including a strong signal from fatty connective tissue and red blood cells. Adenocarcinoma was characterized by loss of SHG signal, whereas cystic abnormalities showed strong SHG. There was limited overlap of two-and three-photon signals, suggesting that three-photon imaging can provide additional information for early diagnosis of ovarian cancer.

One-color resonance two-photo ionization (R2PI) spectra of mixed clusters of styrene molecule (S) with polar molecules [water (W), methanol (M), ethanol (E), and trifuoroethanol (T)] were measured through the S1←S0 transition of the styrene molecule. The spectra reveal a rapid increase in complexity with the number of polar molecules in the cluster, associated with van der Waal modes and isomeric forms. The spectral shifts of the cluster origins from the S1-S0 transition of the bare styrene molecule reflect the nature of the intermolecular interactions within the binary clusters. The obtained R2PI spectra xv were compared with the spectra of the analogous benzene containing clusters. The styrene-water and the styrene-methanol complexes exhibited very different spectral shifts and structures as compared to the benzene-water and benzene-methanol complexes, respectively. The favorable interactions of polar molecules with the olefin group of styrene may explain the strong inhibition effects of exerted by small concentrations of water and alcohols on the cationic polymerization of styrene. Size-specified intracluster proton transfer reactions were observed for mixed clusters of styrene dimer with water, methanol and ethanol molecules. It was proposed that the polar molecules tend to aggregate around the olefin center, which promotes the transfer of the charge from the propagating chain to the hydrogen-bonded polar molecules subcluster. The minimum number of polar molecules required for a proton transfer to take place exothermically depends on the proton affinity of the polar molecules subcluster. The estimated upper limit value for the proton affinity of styrene dimer radical was evaluated based on the energetic of the proton transfer reaction to be ≤ 220.4 kcal/mol. No intracluster reaction was observed for styrene-trifluoroethanol (STn) system. In order to provide a comparison between the styrene and benzene systems, the benzene-ethanol (BEn) and benzene-trifluoroethanol (BTn) clusters were studied by using the R2PI technique via the 6¹0 transition of the benzene molecule. Both dissociative electron transfer and dissociative proton transfer reactions were observed within the BEn clusters, where n = 2 and 3, respectively. Proton transfer reactions were observed following dissociative electron transfer reactions within the (BTn) clusters, where n = 4, to generate the protonated clusters (H+Tn).