Biosystem for the culture and electrical characterisation of epithelial cell tissues

The aim of this work was to develop a microchamber system for performing electrophysiological measurements on epithelial surface on culture membranes in the mm2 range. This miniaturised system permits the use of small quantities of epithelial cells, which in relatively short time grow into a tight layer that can be used in electrophysiological (ion transport) experiments. Availability of cells is an issue, for example when the cells originate from human biopsies or from (expensive!) transgenic mice. Apart from having reduced culture surfaces (for use with scarce biological tissues), there are tremendous advantages in having a cell culture mini-chamber. Our systems are with integrated electrical electrodes, micro-fluidic channels and feed-throughs, making them extremely compact and easy to use, thereby avoiding cell perturbation by manipulations. The structures facilitate control of the cell layer growth, the measurement of the cell layer resistance and the transport and diffusion of biological or pharmacological molecules through the cell layer. Moreover we have chosen cheap and easy-to-tool materials for the realisation of disposable devices. We have also fabricated modular devices, in which the cell culture membranes can be reversibly placed within or removed from the system, thereby offering flexibility and economic interest. These microsystems (for biological applications ("biosystems")) are realised using photolithography, various etching procedures (among which powder-blasting), thin film deposition, electrochemical deposition, polydimethylsiloxane (PDMS) moulding and gluing technologies. Both electrical and fluidic characterisation of the biosystems has been performed. Also, a specific study of microelectrode properties of different electrode materials, such as Pt, Ag and Ag/AgCl has been done using various electrochemical experiments and models. The devices are finally tested in real biological experiments. These experiments were carried on with the collaboration of three different biological academic work groups: the group of Prof. W. Hunziker from the Institute of Biochemistry at the University of Lausanne, the group of Prof. Van der Goot from the Department of Biochemistry at the University of Geneva and Prof. J.D. Horisberger from the Institute of Pharmacology and Toxicology at the University of Lausanne. The functionality of our devices has been tested and their potential for the study of transport and diffusion of biological or pharmacological molecules through the cell layer via accurate measurement of (bio-) chemically induced resistance variations, has been demonstrated.

Electrical and optical properties of Ta-Si-N thin films deposited by reactive magnetron sputtering

David Oezer, Rosendo Sanjines

The electrical and optical properties of TaxSiyNz thin films deposited by reactive magnetron sputtering from individual Ta and Si targets were studied in order to investigate the effects of nitrogen and silicon contents on both properties and their correlation to the film microstructure. Three sets of fcc-TaxSiyNz thin films were prepared: sub-stoichiometric TaxSiyN0.44, nearly stoichiometric TaxSiyN0.5, and over-stoichiometric TaxSiyN0.56. The optical properties were investigated by near-normal-incidence reflectivity and ellipsometric measurements in the optical energy range from 0.375 eV to 6.8 eV, while the d.c. electrical resistivity was measured in the van der Pauw configuration from 20 K to 300 K. The optical and electrical measurements were interpreted using the standard Drude-Lorentz model and the so-called grain boundary scattering model, respectively. The electronic properties were closely correlated with the compositional and structural modifications of the TaxSiyNz films due to variations in the stoichiometry of the fcc-TaNz system and the addition of Si atoms. According to the nitrogen and silicon contents, fcc-TaxSiyNz films can exhibit room temperature resistivity values ranging from 10(2) mu Omega cm to about 6 x 10(4) mu Omega cm. The interpretation of the experimental temperature-dependent resistivity data within the Grain Boundary Scattering model, combined with the results from optical investigations, showed that the mean electron transmission probability G and the free carriers concentration, N, are the main parameters that control the transport properties of these films. The results indicated that the correlation between electrical and optical measurements with the chemical composition and the nanostructure of the TaxSiyNz thin films provides a pertinent and consistent description of the evolution of the Ta-Si-N system from a solid solution to a nanocomposite material due to the addition of Si atoms. (C) 2012 American Institute of Physics. [http://dx.doi.org/10.1063/1.4766904]

Amer Inst Physics2012

Fundamental Applications of Nanopores: Controlled DNA Translocations to Nanofluidics

Sebastian James Davis

Nanopores are nanometer sized openings that are the connection between two electrolyte filled reservoirs. The measurement of the ion transport flowing through such a pore allows to probe physically or biologically interesting phenomena. These range from the passage of biological molecules, to the modulation of current due to multiple physical effects when the nanopore undergoes mechanical strain or pressure induced flow. Many types of nanopores exist: biological protein pores engineered to be able to sequence DNA, glass nanocapillaries easily interfaced with optical tools, or silicon nitride membrane pores which are a standard tool of recent nanotechnology. This thesis is split into two parts. The first focuses on the use of glass nanocapillaries which allow the facile combination of nanopore experiments with optical tweezers. Optical tweezers are a well established single molecule tool that allow precise force measurement on biologically relevant scales. They are used here for the control of DNA passing through the nanopore. Their ability to measure small scale forces allows the detailed investigation of DNA binding proteins, and attempts to measure the force of DNA passage through biological pores. Extensions of these experiments to the elastic behaviour of DNA during its passage through a nanopore reveal the effects of flow generated by the charged surface of nanopores themselves. This motivates attempts to control such flows as well as the second part of the thesis. The second part of the thesis focuses on nanofluidics, the role of fluid flow and ion transport at the nanoscale. Using a setup combining pressure with nanopores it is possible to probe, via precise measurements of the conduction of the nanopore, the wetting state of the pore. Contamination phenomena are shown to be abundant with such small systems and a description of their effects on standard measurements such as direct current current-voltage curves is given. Following this, pressure is applied to perfectly filled pores and, thanks to a new alternating current detection method, is shown to be able to discern the effect of pressure induced strain at the pore as well as the coupling of hydraulic flow with electrical properties of the pore. Finally, extensions beyond aqueous solvents are explored in both nanocapillaries and silicon nitride pores. For this, room-temperature ionic liquids are used. These liquids are known from previous studies to behave differently at surfaces and in nano-confinement. The nanopore system both with and without added pressure is shown to be a good tool for investigating such phenomena.

EPFL2020

Une première mesure lidar combinée d'ozone et de vent, à partir d'une instrumentation et d'une méthodologie coup par coup

During these last decades, air pollution has drawn the attention of scientists because of the deterioration of the human environment that it has caused. Unfortunately, the study of this dynamic phenomenon is very complex and requires the use of models that compute the evolution of the physico-chemical parameters of the atmosphere. Moreover, the predictions of the numerical simulations have to be validated by range- and time-resolved measurements, carried out continuously up to the top of the planetary boundary layer. Only the optical radar or lidar satisfies these specifications: for this reason the Laboratory for Air and Soil Pollution (LPAS) of the Swiss Federal Institute of Technology (EPFL) has developed such a system in order to provide the modellers with information complementary to that given by the ground measurements. This instrument, after mounting on a mobile platform, has determined the concentration of the key molecule ozone during several field campaigns, demonstrating its reliability and precision. Moreover, the EPFL system is a shot per shot lidar, able to record each detected signal and not only its average in a time interval. This feature is innovative to lidar determinations of tropospheric ozone and allows one to measure accurately the statistical error, to correct for the effect of systematic biases and to observe the dynamic behaviour of the atmosphere: the measurement of the statistical error has been used to build a model of the signal and of its noise, useful in the design of future lidar systems and for the comparison of the various data processing algorithms. the correction of the bias caused by the artificial deformation of the statistical distribution of the signal at long distance has led to a considerable increase of the range of the retrieved ozone profiles, the observation of the dynamic behaviour of the atmosphere has resulted in the first simultaneous measurement of a pollutant concentration and of the wind velocity with the same data set recorded by a lidar.

EPFL1996