Thesis (PhD)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: See full text / AFRIKAANSE OPSOMMING: Sien volteks

Optical transfection is a promising technique for the delivery of foreign genetic material into cells by transiently changing the permeability of the cell membrane. Of the different optical light sources that have been used, femtosecond laser based transfection has been one of the most effective methods for optical transfection which is generally implemented using a free-space bulk optical setup. Here in this thesis, a few novel fabrication methods are devised to obtain easy and inexpensive fabrication of microlensed optical fibers, which can be used to replace traditional optical setup and perform femtosecond optical transfection. These fabrication methods offer the flexibility to fabricate a microlens which can focus femtosecond laser pulses at 800 nm to a small focal spot whilst keeping a relatively large working distance. In conventional optical transfection methods the foreign genetic material to be transfected is homogenously mixed in the medium. This thesis reports the first realization of an integrated optical transfection system which can achieve transfection along with localized drug delivery by combining lensed fiber based optical transfection system with a micro-capillary based microfluidic system. Finally, based on an imaging fiber (coherent optical fiber bundle), the first endoscope-like integrated system for optical transfection with subcellular resolution epifluorescence imaging was built. The transfection efficiency of these fiber based systems is comparable to that of a standard free-space transfection system. Also the use of integrated system for localized gene delivery resulted in a reduction of the required amount of genetic material for transfection. The miniaturized, integrated design opens a range of exciting experimental possibilities, such as the dosing of tissue slices to study neuron activities, targeted drug delivery, and in particular for using endoscope-like integrated systems for targeted in vivo optical microsurgery.

This dissertation is on the development of a laser system for the generation of femtosecond frequency combs in the vacuum-ultraviolet (VUV) via intracavity high-harmonic generation (HHG). The HHG process yields coherent vacuum ultraviolet (VUV) light resulting from the ionization of noble gases driven by intense near-IR femtosecond frequency combs in an optical enhancement cavity. An injection locked amplification cavity (fsAC) was developed in order to generate a high power femtosecond frequency combs based on a Ti:Sapphire oscillator. Detailed amplifier performance was investigated in order to evaluate the coherence of the pulse amplification process. A passive power enhancement cavity for fs pulses (fsEC) was designed for intracavity high harmonic generation. For maximum power enhancement and conversion efficiency, the intracavity dispersion was compensated and various design layouts tested. A careful analysis of the phase matching conditions was performed, taking into account the effect of reabsorption of the generated high harmonic light, to compare different cavity geometries and determine which would produce the most efficient harmonic yield. Numerical simulations were also performed to determine the level of intra-cavity ionization that could be sustained before disrupting the pulse enhancement process. Based on the results of these simulations and calculations, it was determined that for a xenon gas target, a moderate peak intensity of the order of ~ 5×10¹³W/cm² produces harmonics most efficiently.

An optical detection technique is developed to detect and measure the resonant vibration of the cellular membrane. Biological membranes are active components of living cells and play a complex and dynamic role in life processes. They are believed to have oscillation modes of frequencies in the range of 1 to 1000 GHz. To measure such a high-frequency vibration, a linear laser cavity is designed to produce a train of femtosecond pulses of adjustable repetition rate. The method is then directly applied to liposomes, "artificial membrane", stained with a liphophilic potential sensitive dye. The spectral behavior of a selection of potential sensitive dyes in the membrane is also studied.

One of the most recent advancements in laser technology is the development of ultrashort pulsed femtosecond lasers (FSLs). FSLs are improving many fields due to their unique extreme precision, low energy and ablation characteristics. In the area of laser medicine, ophthalmic surgeries have seen very promising developments. Some of the most commonly performed surgical operations in the world, including laser-assisted in-situ keratomileusis (LASIK), lens replacement (cataract surgery), and keratoplasty (cornea transplant), now employ FSLs for their unique abilities that lead to improved clinical outcome and patient satisfaction. The application of FSLs in medical therapeutics is a recent development, and although they offer many benefits, FSLs also stimulate nonlinear optical effects (NOEs), many of which were insignificant with previously developed lasers. NOEs can change the laser characteristics during propagation through a medium, which can subsequently introduce unique safety concerns for the surrounding tissues. Traditional approaches for characterizing optical effects, laser performance, safety and efficacy do not properly account for NOEs, and there remains a lack of data that describe NOEs in clinically relevant procedures and tissues. As FSL technology continues to expand towards new applications, FSL induced NOEs need to be better understood in order to ensure safety as FSL medical devices and applications continue to evolve at a rapid pace. In order to improve the understanding of FSL-tissue interactions related to NOEs stimulated during laser beam propagation though corneal tissue, research investigations were conducted to evaluate corneal optical properties and determine how corneal tissue properties including corneal layer, collagen orientation and collagen crosslinking, and laser parameters including pulse energy, repetition rate and numerical aperture affect second and third-harmonic generation (HG) intensity, duration and efficiency. The results of these studies revealed that all laser parameters and tissue properties had a substantial influence on HG. The dynamic relationship between optical breakdown and HG was responsible for many observed changes in HG metrics. The results also demonstrated that the new generation of therapeutic FSLs has the potential to generate hazardous effects if not carefully controlled. Finally, recommendations are made to optimize current and guide future FSL applications.

La texturisation de matériaux par irradiation laser ultracourt est un procédé permettant de modifier les propriétés optiques et électriques de la matière en formant des nano et microstructures en surface, apparaissant au cours des irradiations successives. Le contrôle du procédé et le développement des applications nécessitent une compréhension des mécanismes mis en jeu. Les processus intervenant sont étudiés à l'aide de simulations numériques, et sont comparés à des résultats expérimentaux. L'étude est menée dans le cadre de l'augmentation du rendement des cellules photovoltaïques basées sur du silicium massif. / Ultrashort laser pulsed texturing is a process which allows to modify optical and electrical properties of matter, through formation of nano and micro structures on surface, appearing from pulse to pulse. Control of the process and developments of the potential applications need a good knowledge of the formation mechanisms. Processes occuring during the interaction are studied using numerical simulations and are compared to experimental results. The study aims to increase the efficiency of solar cells based on bulk silicon.

Doctor of Philosophy / Department of Industrial and Manufacturing Systems Engineering / Shuting Lei / Shuting Lei / Femtosecond (fs) laser ablation possesses unique characteristics for micromachining, notably non-thermal interaction with materials, high peak intensity, precision and flexibility. In this dissertation, the potential of fs laser ablation for machining polyurea aerogel and scribing thin film solar cell interconnection grooves is studied. In a preliminary background discussion, some key literature regarding the basic physics and mechanisms that govern ultrafast laser pulse interaction with materials and laser micromachining are summarized. First, the fs laser pulses are used to micromachine polyurea aerogel. The experimental results demonstrate that high quality machining surface can be obtained by tuning the laser fluence and beam scanning speed, which provides insights for micromachining polymers with porous structures. Second, a new fs laser micro-drilling technique is developed to drill micro-holes in stainless steel, in which a hollow core fiber is employed to transmit laser pulses to the target position. The coupling efficiency between the laser and the fiber is investigated and found to be strongly related to pulse energy and pulse duration. Third, the fs laser with various energy, pulse durations, and scanning speeds has been utilized to pattern Indium Tin Oxide (ITO) glass for thin film solar cells. The groove width decreases with increasing pulse duration due to the shorter the pulse duration the more effective of the energy used to material removal. In order to fully remove ITO without damaging the glass, the beam scanning speed need to precisely be controlled. Fourth, fs laser has been utilized to scribe Molybdenum thin film on Polyimide (PI) flexible substrate for Copper Indium Gallium Selenide (CIGS) thin film solar cells. The experimental parameters and results including ablation threshold, single- and multiple-pulse ablation shapes and ablation efficiency were discussed in details. In order to utilize the advantages of the fs lasers, the fabrication process has to be optimized for thin film patterning and structuring applications concerning both efficiency and quality. A predictive 3D Two Temperature Model (TTM) was proposed to predict ablation characteristics and help to understand the fs laser metal ablation mechanisms. 3D temperature field evolution for both electrons and lattice were demonstrated. The ablation model provides an insight to the physical processes occurring during fs laser excitation of metals. Desired processing fluence and process speed regime can be predicted by calculating the ablation threshold, ablation rate and ablation crater geometry using the developed model.

Master of Science / Department of Physics / Matthias Kling / This thesis consists of two major parts. The first part concerns studies of the orientation dependence of the ionization of diatomic molecules in intense, femtosecond two-color laser fields. The second part is about studies on the ionization mechanisms of the C[subscript]6[subscript]0 molecule in femtosecond near-infrared and ultraviolet laser fields. In the first part, experimental and theoretical results on the asymmetric ion emission of the heteronuclear molecules CO and NO in two-color laser fields are discussed. The two-color fields, which can be tailored by a relative phase, are used to ionize and dissociate CO and NO molecules, both of which are molecules with small polarizabilities. The resulting C[superscript]+, C[superscript]2[superscript]+, N[superscript]+ and O[superscript]+ ions are detected by a velocity map imaging (VMI) setup. The photoelectrons from above-threshold ionization (ATI) of Xe are studied under such a two-color field to assign the phase. For both CO and NO we find that enhanced ionization occurs when the molecule is oriented with the electric field pointing from the C or N atom toward the O atom. This is in agreement with the molecular orbital Ammosov-Delone-Krainov (MO-ADK) theory and the Stark-corrected strong-field-approximation (SFA) calculations. The second part is devoted to the investigation of the ionization mechanism of neutral C[subscript]6[subscript]0 molecules with 30 fs laser pulses at about 800 nm and with 50 fs pulses at about 400 nm. The angular distributions of photoelectrons are measured utilizing VMI. Measurements under different intensities are carried out for the two wavelengths. In our work, thermal electron emission is highly suppressed by the use of short pulses. For near-infrared excitation, photoelectron angular distributions (PADs) that contain six lobes are observed for low energy electrons. This behavior is different from studies for longer pulses of about 120 fs [1]. Further analysis indicates that the PADs might originate from single photon ionization of a super atomic molecular orbital (SAMO), however, a detailed assignment requires further theoretical work. The PADs for the ultraviolet excitation show very similar structures to earlier results [1]. For the near-infrared excitation, we have carried out studies as a function of the chirp of the pulses and find effects on photoelectron spectra and on PADs, which are tentatively explained by sequential multiphoton ionization via “doorway” states.

L’Industrie actuelle demande des produits à haute valeur ajoutée offrant des nouvelles fonctions à moindre coût. Parmi les fonctions on peut citer la coloration de surface, le noircissement de surface, la réduction des frottements, la génération de surface anti-réflexion, anti-bactérienne, superhydrophobe ou anti-formation de glace. Les surfaces fonctionnelles présentes dans la nature nous indiquent que ces propriétés uniques sont possibles par des texturations de surface à l’échelle micro et nanométrique adéquates.Parallèlement à cela, la technologie laser révolutionne le champ des possibles en termes de texturation de surface et permet de reproduire ces fonctions inspirées du monde du vivant en modifiant la morphologie et la chimie de surface. Néanmoins, le développement et le déploiement de telles techniques de texturation laser au niveau industriel nécessite la levée de trois verrous. Le premier est de connecter les propriétés macroscopiques (couleur, résistance mécanique, stabilité chimique, vieillissement) et la morphologie de surface aux échelles nano et microscopiques. Le second d’acquérir une parfaite maîtrise de la morphologie de surface à ces échelles. Le troisième est la transposition du procédé développé en laboratoire en procédé industriel adapté aux traitements de grandes surfaces avec des temps de cycles les plus courts possibles. Nous avons étudié plusieurs techniques de texturation de surface à l’échelle submicronique par laser femtoseconde. Ainsi des « ripples » de quelques dizaines de nanomètres ont être réalisées par laser UV. L’irradiation avec double impulsion apporte une capacité supplémentaire dans le contrôle de la morphologie de surface finale. Différents types de structures, avec différentes symétries, ont ainsi été produites en jouant sur le délai entre les deux impulsions. Des structures LIPSS homogènes triangulaires ou carrées ont été obtenues pour des délais inférieurs à 5 ps et 500 ps respectivement. Des paramètres opératoires, en particulier la fluence et la polarisation, ont été identifiés comme jouant un rôle majeur dans les caractéristiques de la morphologie de surface finale. Des expériences complémentaires ont montré que des résultats similaires peuvent être obtenus en utilisant des cristaux biréfringents pour générer des délais courts. Nous avons également exploré la possibilité d’utiliser des trains d’impulsions uniques pour produire des texturations de surface de dimensions caractéristiques supérieures allant de quelques microns à plusieurs dizaines de microns en faisant varier de manière systématique la fluence, la dose énergétique et le taux de répétition du laser. La comparaison de résultats expérimentaux avec ceux issus de simulation nous avons mis en évidence le rôle majeur de l’accumulation thermique sur les dimensions caractéristiques des structures générées par laser. Par ailleurs, nous avons démontré la capacité du procédé à produire de texturations sub-longueurs d’onde, homogènes, sur des surfaces supérieures à 1 cm², avec des lasers ayant des taux de répétitions allant jusqu’à 10 MHz et des systèmes de positionnement innovants. Des nano-rugosités de surface ainsi produites affichent des propriétés de super hydrophobicité. A titre d’exemple, nous avons atteint un temps de texturation de l’ordre de 1 min/cm², soit 60 fois inférieurs à ce que nous obtenions en début des travaux. Enfin, nous avons démontré un temps de 9 s/cm² pour le noircissement de surface.Ces travaux de recherche, mettant à profit des sources laser et des équipements de déflection optique de dernière génération, apportent une contribution significative dans la compréhension des mécanismes d’une part, et dans la capacité à contrôler et à produire de telles texturations sur des grandes surfaces d’autre part. Ils devraient favoriser une dissémination rapide de ces technologies de texturation laser dans l’industrie. / Current industrial markets demand highly value-added products offering new features at a low-cost. Among the most desired functionalities are surface colouring and blackening, anti-icing, anti-biofouling, wear reduction and anti-reflectivity. Laser surface processing holds a virtually endless potential in surface functionalization since it can generate versatile surface properties by modifying surface morphology and chemistry. Nevertheless, developing functional surfaces for implementation in the industry requires action on three levels. The first is to connect the macro-scale surface properties (colour, mechanical resistance, chemical stability, ageing) and the micro & nano-scale morphology. The second is to increase the level of control over the laser induced morphology in the near micron and submicron scale. The third is to upscale the lab-developed process both in terms of processed area and cycle time. Functional textures found in nature can be used as a guideline for connecting the surface texture with the surface property. It is well established that different textures can enable different functionalities. Nevertheless, the level of control of the laser induced morphology has to be improved significantly in order to allow one to mimic nature’s examples. Increase of control requires an in-deep understanding of the physical mechanisms that lead to nanostructure formation. To this end, we carry out a comprehensive parametric study of fs processing on stainless steel. The impact of wavelength, overlap, fluence, dose, repetition rate, polarization and interpulse delay in the induced morphology was investigated.We investigate several techniques to achieve controlled laser structuring in the submicron regime. Ripples of a few tens of nanometres were obtained with a UV laser. Double pulses were employed to further control the submicron structures. Structures of different size and symmetry were obtained in different delays underlining the key role of the interpulse delay (Δτ). Homogeneous triangular and square 2D-LIPSS were obtained for Δτ smaller than 5 ps and 500 ps, respectively. Process parameters and particularly fluence and polarization were found to play also a role in the laser induced feature characteristics. In a complementary set of experiments, we show that similar results can be obtained for small delays with a robust setup of birefringent crystals. In the above micron regime, trains of single pulses were employed for controlling the surface morphology. Fluence, dose and repetition rate, were varied to show a systematic variation of spikes in the range of tens of micrometers. Combining our experimental results with simulation data we underline the key role of heat accumulation on the structures size. Finally, we proposed an upscaling strategy showing the possibility to exploit repetition rates up to 10 MHz for laser texturing.In the upscaling part, areas much larger than the spot size were textured homogenously using high repetition rate laser and innovative laser positioning systems. Nanometric ripples induced by UV laser act as a subwavelength grating. Laser induced nano roughness exhibits superhydrophobic properties. Uniform distribution of well-defined, sub-wavelength, 2D-LIPSS was successfully generated over ~1 cm2. The final surface exhibits multiple axis iridescence giving a holographic effect. Employing a 10 MHz laser surface was textured at a rate of ~ 1 min/cm2 almost 60 times faster compared to our starting point. Lastly, surface blackening was achieved at a rate of ~ 9 sec/cm2.In conclusion, valuable data were provided both in surface functionalization, in understanding and controlling of laser induced structuring and in upscaling a lab developed process. We believe that our results open the way for exploiting fs laser texturing in everyday applications employing up to date laser sources and positioning systems.

A primeira etapa do projeto foi realizar testes para usinagem controlada e otimizada de vidro ótico de borosilicato (BK7) por laser de femtossegundos. Parâmetros como energia, pulsos sobrepostos e a variação da posição focal foram investigados para controle da taxa de remoção do material e extensão da cratera ablacionada. Especial atenção foi dada à condição física e topográfica da superfície resultante da usinagem para torná-la menos rugosa e evitar a retenção de reagentes que possam contaminar e alterar as reações pretendidas. Microcanais, microválvulas, microbombas, misturadores, microrreatores, aquecedores e outros componentes foram desenvolvidos para compor sistemas microfluídicos. Os microcanais construídos sobre a superfície de vidro BK7 vedados por uma lâmina de polidimetilsiloxano (PDMS) são a base dos sistemas microfluídicos. O controle de fluxo de reagentes é feito por miniválvulas pneumáticas controladas por um microcontrolador Arduino através de uma plataforma Labview. Este trabalho mostra os componentes desenvolvidos e dois sistemas microfluídicos criados. O primeiro contém um circuito capaz de replicar ensaios imunoenzimáticos (ELISA) com um custo muito menor de insumos. O segundo é um sistema para a produção de nanocristais fluorescentes de NaYF4 especialmente utilizados como marcadores em imagens de sistemas biológicos. / The first stage of the project was to perform tests for controlled and optimized machining of borosilicate optical glass (BK7) by femtosecond laser. Parameters such as energy, number of overlapped pulses, and the focal position variation were investigated for a better extraction of material. Microchannels, microvalves, micropumps, mixers, reactors, heaters and other components were developed to compose applied microfluidic systems. Microchannels built on the surface of BK7 glass sealed by a polydimethylsiloxane (PDMS) sheet form the basis of the microfluidic circuits. The reagents flow control is done by pneumatic mini-valves controlled by an Arduino microcontroller through a Labview platform. This work shows the components developed and two microfluidic systems created. The first contains a microfluidic circuit capable of replicating enzyme-linked immunosorbent assays (ELISA) with a much lower cost of materials. The second has a microfluidic circuit for the production of NaYF4 fluorescent nanocrystals specially used as markers in images of biologic systems.