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<p>Understanding and predicting chemical phenomena is the main goal of computational chemistry. In this thesis I present my work on applying computational approaches to study chemical processes in biological and supramolecular systems.</p> <p>pH-responsive molecular tweezers have been proposed as an approach for targeting drug-delivery to tumors, which tend to have a lower pH than normal cells. In chapter 2 I present a computational study I performed on a pH-responsive molecular tweezer using <em>ab initio</em> quantum chemistry in the gas phase and molecular dynamics simulations in solution. The binding free energy in solution was calculated using Steered Molecular Dynamics. We observe, in atomistic detail, the pH-induced conformational switch of the tweezer and the resulting release of the drug molecule. Even when the tweezer opens, the drug molecule remains near a hydrophobic arm of the molecular tweezer. Drug release cannot occur, it seems, unless the tweezer is a hydrophobic environment with low pH.</p> <p>The protonation state of amino acid residues in proteins depends on their respective pK_a values. Computational methods are particularly important for estimating the pK_a values of buried and active site residues, where experimental data is scarce. In chapter 3 I used the cluster model approach to predict the pK_a of some challenging protein residues and for which methods based on the numerical solution of the Poisson-Boltzmann equation and empirical approaches fail. The ionizable residue and its close environment were treated quantum mechanically, while the rest of the protein was replaced by a uniform dielectric continuum. The approach was found to overestimate the electrostatic interaction leading to predicting lower pK_a values.</p> / Master of Science (MSc)

Ce travail de thèse s’inscrit dans les efforts actuellement réalisés, pour améliorer l’efficacité des pinces optiques conventionnelles qui permettent de manipuler sans contact des objets de quelques dizaines de nanomètres à quelques dizaines de micromètres avec une extrême précision et trouvent de nombreuses applications en biophysique et sciences de colloïdes.L’objectif de cette thèse a été d’explorer une nouvelle approche pour la réalisation de Nanopinces Optiques. Elle s’appuie sur l’utilisation de cavités à cristaux photoniques à modes de Bloch lents. Ces cavités peuvent être efficacement et facilement excitées par un faisceau Gaussien à incidence normale. Contrairement aux pinces optiques conventionnelles, des objectifs à faibles ouvertures numériques peuvent être utilisés. Les performances attendues en termes de piégeage vont bien au-delà de limitations imposées par la limite de diffraction pour les pinces conventionnelles. Ce travail démontre expérimentalement l’efficacité de l’approche. Cette thèse comporte deux parties principales. Dans un premier temps, il a fallu monter un banc expérimental pour mener nos études. Nous avons construit un banc optique, interfacé les instruments, et développé des applications logicielles pour analyser les données. Deux éléments importants ont présidé à sa construction : - Le développement d’un système optique permettant d’exciter les nanostructures photoniques - la conception d’un système d’imagerie pour suivre les nanoparticules. La seconde partie de ce travail a porté sur la mise en évidence du piégeage optique à l’aide de nanostructure à base de cristaux photonique. Nous avons d’abord montré que même des cavités possédant des coefficients de qualités modérés (quelques centaines) permettait d’obtenir des pièges optiques dont l’efficacité est d’un ordre de grandeur supérieur à celui de pinces conventionnels. Fort de ce résultat, nous avons exploré un nouveau type de cavité à cristaux photoniques s’appuyant sur une approche originale : des structures bi-périodiques. Nous avons montré qu’à l’aide de cette approche des facteurs de qualités de l’ordre de plusieurs milliers étaient facilement atteignable. A l’aide de ces nouvelles structures, nous sommes arrivés aux résultats le plus important de ce travail : le piégeage de nanoparticules de 250nm de rayon avec une puissance optique incidente de l’ordre du milliwatt. Une analyse fine du mouvement de la nanoparticule, nous a permis de trouver la signature du mode de Bloch lent. / This thesis aims at improving the efficiency of conventional optical tweezers (cOT). They allow to manipulate objects with dimension from a few tens of nanometer to a few tens of micrometers with a high accuracy and without contact. This has numerous applications in biophysics and colloidal science. This thesis investigates a new approach for optical nanotweezers. It uses a photonic crystal (PC) cavity which generates a slow Bloch mode. This cavity can be effectively and easily excited with a Gaussian beam at the normal incidence. Contrarily to cOT, objective with a small numerical aperture can be used. The expected performances in terms of trapping go well beyond the diffraction limit of cOT. This work demonstrates experimentally the efficacy of approach. This thesis is divided in two main sections. First, we had to set up an experimental bench to carry out to our study. We built the optical bench interface instruments and develop programs to analyze the data. Two essential elements have been considered: - The development of the optical system allowing the excitation of the photonics nanostructure. - The design an imaging system to track nanoparticles. Second, we have focus on the demonstration of the optical trapping. We started by with a low Q factor (few hundred) cavity. Trapping efficiency of an order of magnitude higher than cOT has been demonstrated. Then, we have explored a new king of PC cavity based on double period structure. We show that thanks to this approach high Q factor of several thousand are easily reached. With this structure, we managed to trap 250nm polystyrene beads, with an optical power of the order of a milliwatt. A deep analysis of the nanoparticle trajectories allowed us to find a slow Bloch mode signature.

Integration and miniaturisation in electronics has undoubtedly revolutionised the modern world. In biotechnology, emerging lab-on-a-chip (LOC) methodologies promise all-integrated laboratory processes, to perform complete biochemical or medical synthesis and analysis encapsulated on small microchips. The integration of electrical, optical and physical sensors, and control devices, with fluid handling, is creating a new class of functional chip-based systems. Scaled down onto a chip, reagent and sample consumption is reduced, point-of-care or in-the-field usage is enabled through portability, costs are reduced, automation increases the ease of use, and favourable scaling laws can be exploited, such as improved fluid control. The capacity to manipulate single cells on-chip has applications across the life sciences, in biotechnology, pharmacology, medical diagnostics and drug discovery. This thesis explores multiple applications of optical manipulation within microfluidic chips. Used in combination with microfluidic systems, optics adds powerful functionalities to emerging LOC technologies. These include particle management such as immobilising, sorting, concentrating, and transportation of cell-sized objects, along with sensing, spectroscopic interrogation, and cell treatment. The work in this thesis brings several key applications of optical techniques for manipulating and porating cell-sized microscopic particles to within microfluidic chips. The fields of optical trapping, optical tweezers and optical sorting are reviewed in the context of lab-on-a-chip application, and the physics of the laminar fluid flow exhibited at this size scale is detailed. Microfluidic chip fabrication methods are presented, including a robust method for the introduction of optical fibres for laser beam delivery, which is demonstrated in a dual-beam optical trap chip and in optical chromatography using photonic crystal fibre. The use of a total internal reflection microscope objective lens is utilised in a novel demonstration of propelling particles within fluid flow. The size and refractive index dependency is modelled and experimentally characterised, before presenting continuous passive optical sorting of microparticles based on these intrinsic optical properties, in a microfluidic chip. Finally, a microfluidic system is utilised in the delivery of mammalian cells to a focused femtosecond laser beam for continuous, high throughput photoporation. The optical injection efficiency of inserting a fluorescent dye is determined and the cell viability is evaluated. This could form the basis for ultra-high throughput, efficient transfection of cells, with the advantages of single cell treatment and unrivalled viability using this optical technique.

Cette thèse traite de la manipulation d'atomes de rubidium 87 piégés dans un piège optique dipolaire microscopique. Les expériences sont réalisées dans divers régimes de chargement du piège allant d'un atome unique à quelques milliers d'atomes en moyenne.Le régime à un seul atome permet de calibrer le dispositif expérimental. Nous utilisons l'atome unique comme bit quantique dont nous pouvons préparer et lire l'état avec une efficacité de 99.97% et 98.6%, respectivement. Lorsque plusieurs atomes sont chargés dans le piège microscopique, nous observons une distribution sub-Poissonienne du nombre d'atomes, liée aux collisions assistées par la présence de lumière quasi résonante. Une étude de ces collisions dans notre cas particulier (piège microscopique) révèle des taux de pertes extrêmement élevés, proches de la limite théorique de Langevin. Enfin, nous montrons que le chargement du piège microscopique avec plusieurs atomes est plus efficace lorsque nous superposons sur ce piège un deuxième piège, macroscopique, qui joue le rôle de réservoir d'atomes. Ce réservoir permet de charger le micro-piège à partir du macro-piège en l'absence de lumière quasi résonante et donc d'éviter les collisions assistées par la lumière.Le chargement du micro-piège à partir du macro-piège conduit à des conditions initiales optimales pour l'évaporation forcée dans la perspective d'atteindre la condensation de Bose-Einstein avec seulement une dizaine d'atomes. Après évaporation du gaz nous atteignons des densités dans l'espace des phases proches du régime de dégénérescence. / This thesis focuses on the manipulation of rubidium 87 atoms in a microscopic optical dipole trap. The experiments are performed in various regimes where the number of atoms in the microscopic trap ranges from exactly one atom to several thousands on average.The single atom regime allows us to calibrate the experimental setup. We use it a quantum bit, which state we can prepare and read out with efficiencies of 99.97% and 98.6%, respectively. When several atoms are loaded in the microscopic trap we observe a sub-Poissonian distribution of the number of atoms due to light-assisted collisions in the presence of near-resonant light. A study of these collisions in our particular case (microscopic trap) reveals extremely high loss rates approaching the theoretical Langevin limit. Finally, we demonstrate that the loading of the microscopic trap is more efficient when we superimpose on this trap a second macroscopic trap, which we use as an atom reservoir. This reservoir allows us to load the micro trap from the macro trap in the absence of any near-resonant light, thus avoiding light-assisted collisions.The loading of the micro trap from the macro trap leads to optimal initial conditions for forced evaporation towards Bose-Einstein condensation with about ten atoms only. After evaporation we reach phase-space densities approaching the degenerate regime.

Les atomes neutres sont des candidats prometteurs pour la réalisation et l’étude d’états intriqués à quelques dizaines de particules. Pour générer de tels états, une approche consiste à utiliser le mécanisme de blocage dipolaire résultant des fortes interactions dipôle-dipôle entre atomes de Rydberg.Suivant cette approche, cette thèse présente la conception et la caractérisation d’un dispositif expérimental permettant de manipuler des atomes de 87Rb individuels piégés dans des pinces op- tiques microscopiques, et à les exciter vers des états de Rydberg. Un environnement électrostatique stable et des électrodes de contrôle permettent une manipulation fine de ces états. Avec deux pinces optiques séparées de quelques microns, nous démontrons le blocage de Rydberg entre deux atomes, et nous observons leur excitation collective.Enfin, en opérant en régime de blocage partiel, nous développons une méthode permettant de mesurer l’interaction de van der Waals ∆E = C6 /R6 entre deux atomes séparés par une distance R contrôlée. Les coefficients C6 obtenus pour différents états de Rydberg sont en bon accord avec des calculs théoriques ab initio, et nous observons l’augmentation spectaculaire de l’interaction en fonction du nombre quantique principal n de l’état de Rydberg. / Neutral atoms are promising candidates for the realization of entangled states involving up to a few tens of particles. To generate such states, one approach consists in using the dipole blockade mechanism, which results from the strong dipole-dipole interactions between Rydberg atoms.Following this approach, this thesis describes the design and the characterization of an experimental apparatus allowing to manipulate single 87Rb atoms trapped in microscopic optical tweezers, and to excite them towards Rydberg states. A stable electrostatic environment and controlled electrodes enable the fine manipulation of these states. Using two optical tweezers separated by a few microns, we demonstrate the Rydberg blockade between two single atoms, and we observe their collective excitation.Finally, by operating in the partial blockade regime, we develop a method allowing to measure the van der Waals interaction ∆E = C6 /R6 between two atoms separated by a controlled distance R. The C6 coefficients obtained for various Rydberg states agree well with ab initio theoretical calculations, and we observe the dramatic increase of the interaction with the principal quantum number n of the Rydberg state.

This thesis describes an experimental study on fluctuations of a Brownian particle immersed in a fluid, confined by optical tweezers and subject to two different kinds of non-equilibrium conditions. We aim to gain a rather general understanding of the relation between spontaneous fluctuations, linear response and total entropy production for processes away from thermal equilibrium. The first part addresses the motion of a colloidal particle driven into a periodic non-equilibrium steady state by a nonconservative force and its response to an external perturbation. The dynamics of the system is analyzed in the context of several generalized fluctuation-dissipation relations derived from different theoretical approaches. We show that, when taking into account the role of currents due to the broken detailed balance, the theoretical relations are verified by the experimental data. The second part deals with fluctuations and response of a Brownian particle in two different aging baths relaxing towards thermal equilibrium: a Laponite colloidal glass and an aqueous gelatin solution. The experimental results show that heat fluxes from the particle to the bath during the relaxation process play the same role of steady state currents as a non-equilibrium correction of the fluctuation-dissipation theorem. Then, the present thesis provides evidence that the total entropy production constitutes a unifying concept which links the statistical properties of fluctuations and the linear response function for non-equilibrium systems either in stationary or non stationary states.

Each organism has to maintain the integrity of its genetic code, which is stored in its DNA. This is achieved by strongly controlled and regulated cellular processes such as DNA replication, -repair and -recombination. An essential element of these processes is the unwinding of the duplex strands of the DNA helix. This biochemical reaction is catalyzed by helicases that use the energy of nucleoside triphophate (NTP) hydrolysis. Although all helicases comprise highly conserved domains in their amino acid sequence, they exhibit large variations regarding for example their structure, their function and their target nucleic acid structures. The main objective of this thesis is to obtain insight into the DNA unwinding mechanisms of three helicases from two different organisms. These helicase vary in their structures and are involved in different pathways of DNA metabolism. In particular the replicative, hexameric helicase Large Tumor-Antigen (T-Antigen) from Simian virus 40 and the DNA repair helicases RecQ2 and RecQ3 from Arabidopsis thaliana are studied. To observe DNA unwinding by these helicases in real-time on the single molecule level, a biophysical technique, called magnetic tweezers, was applied. This technique allows to stretch single DNA molecules attached to magnetic particles. Simultaneously one can measure the DNA end-to-end distance. Special DNA hairpin templates allowed to characterize different parameters of the DNA unwinding reaction such as the unwinding velocity, the length of unwound DNA (processivity) or the influence of forces. From this mechanistic models about the functions of the helicases could be obtained. T-Antigen is found to be one of the slowest and most processive helicases known so far. In contrast to prokaryotic helicases, the unwinding velocity of T-Antigen shows a weak dependence on the applied force. Since current physical models for the unwinding velocity fail to describe the data an alternative model is developed. The investigated RecQ helicases are found to unwind and close short stretches of DNA in a repetitive fashion. This activity is shown for the first time under external forces. The experiments revealed that the repetitive DNA unwinding is based on the ability of both enzymes to switch from one single DNA strand to the other. Although RecQ2 and RecQ3 perform repetitive DNA unwinding, both enzymes differ largely in the measured DNA unwinding properties. Most importantly, while RecQ2 is a classical helicase that unwinds DNA, RecQ3 mostly rewinds DNA duplexes. These different properties may reflect different specific tasks of the helicases during DNA repair processes. To obtain high spatial resolution in DNA unwinding experiments, the experimental methods were optimized. An improved and more stable magnetic tweezers setup with sub-nanometer resolution was built. Additionally, different methods to prepare various DNA templates for helicase experiments were developed. Furthermore, the torsional stability of magnetic particles within an external field was investigated. The results led to selection rules for DNA-microsphere constructs that allow high resolution measurements. / Jeder Organismus ist bestrebt, die genetischen Informationen intakt zu halten, die in seiner DNA gespeichert sind. Dies wird durch präzise gesteuerte zelluläre Prozesse wie DNA-Replikation, -Reparatur und -Rekombination verwirklicht. Ein wesentlicher Schritt ist dabei das Entwinden von DNA-Doppelsträngen zu Einzelsträngen. Diese chemische Reaktion wird von Helikasen durch die Hydrolyse von Nukleosidtriphosphaten katalysiert. Obwohl bei allen Helikasen bestimmte Aminosäuresequenzen hoch konserviert sind, können sie sich in Eigenschaften wie Struktur, Funktion oder DNA Substratspezifität stark unterscheiden. Gegenstand der vorliegenden Arbeit ist es, die Entwindungsmechanismen von drei verschieden Helikasen aus zwei unterschiedlichen Organismen zu untersuchen, die sich in ihrer Struktur sowie ihrer Funktion unterscheiden. Es handelt sich dabei um die replikative, hexamerische Helikase Large Tumor-Antigen (T-Antigen) vom Simian-Virus 40 und die DNA-Reparatur-Helikasen RecQ2 und RecQ3 der Pflanze Arabidopsis thaliana. Um DNA-Entwindung in Echtzeit zu untersuchen, wird eine biophysikalische Einzelmolekültechnik, die \"Magnetische Pinzette\", verwendet. Mit dieser Technik kann man ein DNA-Molekül, das an ein magnetisches Partikel gebunden ist, strecken und gleichzeitig dessen Gesamtlänge messen. Mit speziellen DNA-Konstrukten kann man so bestimmte Eigenschaften der Helikasen bei der DNA-Entwindung, wie z.B. Geschwindigkeit, Länge der entwundenen DNA (Prozessivität) oder den Einfluß von Kraft, ermitteln. Es wird gezeigt, dass T-Antigen eine der langsamsten und prozessivsten Helikasen ist. Im Gegensatz zu prokaryotischen Helikasen ist die Entwindungsgeschwindigkeit von T-Antigen kaum kraftabhängig. Aktuelle Modelle sagen dieses Verhalten nicht vorraus, weshalb ein alternatives Modell entwickelt wird. Die untersuchten RecQ-Helikasen zeigen ein Entwindungsverhalten bei dem permanent kurze Abschnitte von DNA entwunden und wieder zusammengeführt werden. Dieses Verhalten wird hier zum ersten Mal unter dem Einfluß externer Kräfte gemessen. Es wird gezeigt, dass die permanente Entwindung auf die Fähigkeit beider Helikasen, von einem einzelen DNA-Strang auf den anderen zu wechseln, zurückzuführen ist. Obwohl RecQ2 und RecQ3 beide das Verhalten des permanenten Entwindens aufzeigen, unterscheiden sie sich stark in anderen Eigenschaften. Der gravierendste Unterschied ist, dass RecQ2 wie eine klassische Helikase die DNA entwindet, während RecQ3 eher bestrebt ist, die DNA-Einzelstränge wieder zusammenzuführen. Die unterschiedlichen Eigenschaften könnten die verschieden Aufgaben beider Helikasen während DNA-Reparaturprozessen widerspiegeln. Weiterhin werden die experimentellen Methoden optimiert, um möglichst hohe Auflösungen der Daten zu erreichen. Dazu zählen der Aufbau einer verbesserten und stabileren \"Magnetischen Pinzette\" mit sub-nanometer Auflösung und die Entwicklung neuer Methoden, um DNA Konstrukte herzustellen. Außerdem wird die Torsions\\-steifigkeit von magnetischen Partikeln in externen magnetischen Feldern untersucht. Dabei finden sich Auswahlkriterien für DNA-gebundene magnetische Partikel, durch die eine hohe Auflösung erreicht wird.

In the past decades, numerous single-molecule techniques have been developed to investigate individual bio-molecules and cellular machines. While a lot is known about the structure, localization, and interaction partners of such molecules, much less is known about their mechanical properties. To investigate the weak, non-covalent interactions that give rise to the mechanics of and between proteins, an instrument capable of resolving sub-nanometer displacements and piconewton forces is necessary. One of the most prominent biophysical tool with such capabilities is an optical tweezers. Optical tweezers is a non-invasive all-optical technique in which typically a dielectric microsphere is held by a tightly focused laser beam. This microsphere acts like a microscopic, three-dimensional spring and is used as a handle to study the biological molecule of interest. By interferometric detection methods, the resolution of optical tweezers can be in the picometer range on millisecond time scales. However, on a time scale of seconds—at which many biological reactions take place—instrumental noise such as thermal drift often limits the resolution to a few nanometers. Such a resolution is insufficient to resolve, for example, the ångstrom-level, stepwise translocation of DNA-binding enzymes corresponding to distances between single basepairs of their substrate. To reduce drift and noise, differential measurements, feedback-based drift stabilization techniques, and ‘levitated’ experiments have been developed. Such methods have the drawback of complicated and expensive experimental equipment often coupled to a reduced throughput of experiments due to a complex and serial assembly of the molecular components of the experiments. We developed a high-resolution optical tweezers apparatus capable of resolving distances on the ångstrom-level over a time range of milliseconds to 10s of seconds in surface-coupled assays. Surface-coupled assays allow for a higher throughput because the molecular components are assembled in a parallel fashion on many probes. The high resolution was a collective result of a number of simple, easy-to-implement, and cost-efficient noise reduction solutions. In particular, we reduced thermal drift by implementing a temperature feedback system with millikelvin precision—a convenient solution for biological experiments since it minimizes drift in addition to enabling the control and stabilization of the experiment’s temperature. Furthermore, we found that expanding the laser beam to a size smaller than the objective’s exit pupil optimized the amount of laser power utilized in generating the trapping forces. With lower powers, biological samples are less susceptible to photo-damage or, vice versa, with the same laser power, higher trapping forces can be achieved. With motorized and automated procedures, our instrument is optimized for high-resolution, high-throughput surface-coupled experiments probing the mechanics of individual biomolecules. In the future, the combination of this setup with single-molecule fluorescence, super-resolution microscopy or torque detection will open up new possibilities for investigating the nanomechanics of biomolecules.

Les transitions de phase près d'un point critique - dites du second ordre - sont un sujet toujours d'actualité en raison des nombreux phénomènes critiques intéressants tels que la force de Casimir critique, les problèmes de confinements ou les phénomènes hors d'équilibre suivant une trempe au point critique. Cette thèse vise à étudier expérimentalement certains phénomènes engendrés près d'un point critique. La thèse est divisée en deux axes : le premier consiste à développer plusieurs systèmes expérimentaux qui permettront de mesurer essentiellement la viscosité, par l'intermédiaire des fluctuations thermiques à l'échelle micrométrique. Le deuxième axe consiste à trouver et caractériser des mélanges binaires présentant une transition de phase du second ordre dans lesquelles on souhaite faire des mesures. Les enjeux de ces systèmes expérimentaux sont d'avoir une régulation en température précise, une sonde de mesure sensible aux fluctuations thermiques et/ou à des forces de l'ordre du pN, et un échantillon fiable et reproductible présentant un point critique accessible expérimentalement. Nous avons ainsi monté à partir d'un microscope à force atomique (AFM) déjà présent au laboratoire, un système de mesure de viscosité à sonde AFM fibrée. Malgré sa faible efficacité en terme de sonde de mesure métrologique, nous avons pu décrire et développer un modèle de couplage de modes de vibration permettant de comprendre la mécanique de microleviers AFM fibrés. J'ai également développé au laboratoire la mesure de microscopie dynamique différentielle qui permet de faire des mesures à sondes multiples contrairement au premier montage. J'ai discuté de la précision de la mesure dans le cadre de notre objectif d'étude des fluctuations critiques. En ce qui concerne l'échantillon de mesure, nous avons étudié plusieurs mélanges binaires que nous avons caractérisés par des méthodes classiques de turbidité et diffusion statique de la lumière. Cette caractérisation nous a permis de connaître les mélanges binaires pour les utiliser dans un troisième système de mesure : billes micrométriques piégées dans des pinces optiques déjà monté au laboratoire. Nous y avons rajouté un système de régulation thermique fait maison pour être exploité avec les contraintes de la pince optique. Ces tests ont fait apparaître un phénomène inattendu d'oscillations de transition de phase induites par laser. Nous avons développé un modèle pour les décrire. Enfin, des expériences préliminaires - toujours avec les pinces optiques dans les mélanges binaires - nous ont permis d'observer qualitativement des effets de l'approche au point critique par des mesures de viscosité et d'interaction type force de Casimir critique. / Phase transitions near a critical point, or second order phase transitions, are still a recent object of studies because of the large amount of interesting critical phenomena as the critical Casimir force, confinements problems or out of equilibrium phenomena following a quench at the critical point. This thesis experimentally studies phenomena near a critical point. This manuscript is divided in two parts : the first one consists in building several experimental set-up which measure viscosity through thermal fluctuation at micrometric scale. The second part consists in finding and characterize binary mixtures which show a second order phase transition. Preliminary results have been done in these samples. One of the principal points of these experimental set-up are a well regulated temperature, a probe sensitive to thermal fluctuation and/or pN forces and a reproducible binary mixture which presents a critical point easy to reach experimentally. We mounted from an Atomic Force Microscope (AFM) already built in the laboratory, a hanging-fiber probe to measure viscosity of liquids. Despite its weak efficiency as a metrologic probe, we described and developed a mode coupling model which let us understand mechanics of hanging-fiber probes. I also developed in the lab the dynamic differential microscopy technique (DDM) which do measurements with several probes. I discussed about the measure precision with in mind the aim of studying critical fluctuations. For the choice of the sample, we studied several binary mixtures. We characterized them by classical methods as turbidity measurements and static light scattering. These characterizations let us learn about binary mixtures in order to use them in a third experimental set-up : beads trapped in an optical tweezers already built in the lab. We added to it a home-made thermal regulation which can be used with the constraints of optical tweezers. These tests showed an unexpected phenomenon of oscillating phase transition induce by laser. We developed a model to describe it. At last, preliminary experiments with optical tweezers in binary mixtures showed qualitative effects of an approach near a critical point on the viscosity and on interactions between beads as critical Casimir force.