Determining mammalian cells state by fractal micromotion.

We applied four fractal dimension estimation algorithms on the temporal electrical impedance signal of normal MDCK type II cell cultures monitored by ECIS technique and showed that the fractal dimension due to micromotion allows discriminating processes not sensed by the spectral impedance of the culture. In this work we subjected cell cultures to electric current damage and drug exposure to analyze the changes in the fractal structure of the temporal signal. Among the changes presented and detected are the differentiation between a healthy monolayer and one exposed to a drug, as well as the distinction between a seeding process and a wound-healing process performed by electric current. The four algorithms used were validated by applying them on topological functions of known fractal dimension, a study that determined the necessary conditions for a correct estimation.

Electrical cell impedance spectral mesoscopic model applied to experimental data of variable size microelectrodes.

We apply the electric cell-substrate impedance sensing (ECIS) technique to monolayers of Madin-Darby canine kidney type II cells cultured on microelectrodes of different sizes. We analyze the effect of the microelectrode radius on the parameters provided by existing ECIS models. The cellular properties inferred from the models should be invariant to the change in the microelectrode radius used for the measurements, since these properties are inherent to the type of cells studied. The current standard model, the Giaever-Keese (GK) model, derived from electrical balances of a single cell extended to infinity by suitable boundary conditions, assumes an infinite microelectrode. The model is fitted to experimental data acquired with a large-radius microelectrode, which can be considered infinite for practical purposes. We compute the impedance of the other cell-covered microelectrodes from the parameters obtained with the GK model, resulting in values strongly discrepant with the experimental data for small microelectrodes. We repeat the process with the mean field (MF) model, an alternative model that depends on the microelectrode radius but not on the cell radius. In this paper we introduce the mesoscopic model, an analytical model that simultaneously includes the properties of an individual cell and the sizes of the microelectrode and the insulator (region between the microelectrode and the ground). The impedances calculated with the mesoscopic model are in excellent agreement with experimental data. Finally, the mesoscopic model reduces to the MF model when the insulator goes to infinity and to the GK model when it goes to zero.

Path suppression of strongly collapsing bubbles at finite and low Reynolds numbers.

We study, numerically and experimentally, three different methods to suppress the trajectories of strongly collapsing and sonoluminescent bubbles in a highly viscous sulfuric acid solution. A new numerical scheme based on the window method is proposed to account for the history force acting on a spherical bubble with variable radius. We could quantify the history force, which is not negligible in comparison with the primary Bjerknes force in this type of problem, and results are in agreement with the classical primary Bjerknes force trapping threshold analysis. Moreover, the present numerical implementation reproduces the spatial behavior associated with the positional and path instability of sonoluminescent argon bubbles in strongly gassed and highly degassed sulfuric acid solutions. Finally, the model allows us to demonstrate that spatially stationary bubbles driven by biharmonic excitation could be obtained with a different mode from the one used in previous reported experiments.

Effect of the Rayleigh-Taylor instability on maximum reachable temperatures in laser-induced bubbles.

Laser-induced bubbles provide an effective vehicle to achieve high-energy concentrations and maximum temperatures in bubble luminescence phenomena. One limitation to the temperatures that can be achieved is the development of the Rayleigh-Taylor instability (RTI) during the instants previous to the bubble maximum compression. For a given fluid, the control parameters of the experiment are: the bubble maximum radius, the bubble ambient radius, the initial perturbations of the bubble, and the liquid pressure at infinity. In this work, experiments using laser-induced bubbles in a highly viscous phosphoric acid were performed in order to determine the achievable parameters values in the phase space. The effect of R(max), R(0), a(2)(i), a(3)(i), and p(∞) on the maximum temperature achieved by the gas contents inside the bubble were numerically determined. The results show for each static pressure an optimum region for maximum temperatures of the gas contents bounded by the RTI.

A new experimental device to evaluate eye ulcers using a multispectral electrical impedance technique.

We present a novel experimental technique to determine eye ulcers in animals using a spectral electrical impedance technique. We expect that this technique will be useful in dry eye syndrome. We used a sensor that is basically a platinum (Pt) microelectrode electrically insulated by glass from a cylindrical stainless steel counter-electrode. This sensor was applied to the naked eye of New Zealand rabbits (2.0-3.5 kg in weight). Whereas half of the eyes were normal (control), we applied to the remainder a few drops of 20% (v/v) alcohol to produce an ulcer in the eye. Using a multispectral electrical impedance system we measured ulcerated and control eyes and observed significant difference between normal and pathological samples. We also investigated the effects of different applied pressures and natural degradation of initially normal eyes as a function of time. We believe that this technique could be sufficiently sensitive and repetitive to help diagnose ocular surface diseases such as dry eye syndrome.

Effects of osmolarity on human epithelial conjunctival cells using an electrical technique.

BACKGROUND: The purpose of this study is to report the effect of different media osmolarity on a cell line monolayer of normal human conjunctival epithelia (IOBA-NHC) using Electric Cell-substrate Impedance Sensing (ECIS). METHODS: We built our own ECIS system. We fabricated biocompatible microelectrodes. We used a monolayer of IOBA-NHC cells with media at different osmolarities (315, 360, 446, and 617 mOsm/l). RESULTS: When there is an increase in hyperosmolarity, there is a slight decrease in the measured resistance of the naked microelectrode (without cells), whereas its capacitance remained practically unchanged. The evaluation of resistance and capacitance of a microelectrode covered by a monolayer of IOBA-NHC in relation to a naked microelectrode showed no difference in the standard media (315 mOsm/l), a small difference with 360 mOsm/l, and significant differences with hyperosmolarities of 446 mOsm/l and 610 mOsm/l. The resistance with a confluent cell monolayer is up to three times greater compared to the value of the resistance of the naked electrode with standard media. CONCLUSIONS: Both resistance and capacitance measurements for the cell monolayer were sensitive to changes in osmolarity.

Analytical study of the acoustic field in a spherical resonator for single bubble sonoluminescence.

The acoustic field in the liquid within a spherical solid shell is calculated. The proposed model takes into account Stoke's wave equation in the viscous fluid, the membrane theory to describe the solid shell motion and the energy loss through the external couplings of the system. A point source at the resonator center is included to reproduce the acoustic emission of a sonoluminescence bubble. Particular calculations of the resulting acoustic field are performed for viscous liquids of interest in single bubble sonoluminescence. The model reveals that in case of radially symmetric modes of low frequency, the quality factor is mainly determined by the acoustic energy flowing through the mechanical coupling of the resonator. Alternatively, for high frequency modes the quality factor is mainly determined by the viscous dissipation in the liquid. Furthermore, the interaction between the bubble acoustic emission and the resonator modes is analyzed. It was found that the bubble acoustic emission produces local maxima in the resonator response. The calculated amplitudes and relative phases of the harmonics constituting the bubble acoustic environment can be used to improve multi-frequency driving in sonoluminescence.

Dynamics of sonoluminescing bubbles within a liquid hammer device.

We studied the dynamics of a single sonoluminescing bubble (SBSL) in a liquid hammer device. In particular, we investigated the phosphoric acid-xenon system, in which pulses up to four orders of magnitude brighter than SBSL in water systems (about 10;{12} photons per pulse) have been previously reported [Chakravarty, Phys. Rev. E 69, 066317 (2004)]. We used stroboscopic photography and a Mie scattering technique in order to measure the radius evolution of the bubbles. Under adequate conditions we may position a bubble at the bottom of the tube (cavity) and a second bubble trapped at the middle of the tube (upper bubble). During its collapse, the cavity produces the compression of the liquid column. This compression drives impulsively the dynamics of the upper bubble. Our measurements reveal that the observed light emissions produced by the upper bubble are generated at its second collapse. We employed a simple numerical model to investigate the conditions that occur during the upper bubble collapse. We found good agreement between numerical and experimental values for the light intensity (fluence) and light pulse widths. Results from the model show that the light emission is increased mainly due to an increase in noble gas ambient radius and not because the maximum temperature increases. Even for the brightest pulses obtained ( 2x10;{13} photons, about 20W of peak power) the maximum temperatures computed for the upper bubble are always lower than 20000K .

Experimental study of transient paths to the extinction in sonoluminescence.

An experimental study of the extinction threshold of single bubble sonoluminescence in an air-water system is presented. Different runs from 5% to 100% of air concentrations were performed at room pressure and temperature. The intensity of sonoluminescence (SL) and time of collapse (t(c)) with respect to the driving were measured while the acoustic pressure was linearly increased from the onset of SL until the bubble extinction. The experimental data were compared with theoretical predictions for shape and position instability thresholds. It was found that the extinction of the bubble is determined by different mechanisms depending on the air concentration. For concentrations greater than approximately 30%-40% with respect to the saturation, the parametric instability limits the maximum value of R(0) that can be reached. On the other hand, for lower concentrations, the extinction appears as a limitation in the time of collapse. Two different mechanisms emerge in this range, i.e., the Bjerknes force and the Rayleigh-Taylor instability. The bubble acoustic emission produces backreaction on the bubble itself. This effect occurs in both mechanisms and is essential for the correct prediction of the extinction threshold in the case of low air dissolved concentration.

Trapping an intensely bright, stable sonoluminescing bubble.

Previous works on single bubble sonoluminescence in sulfuric acid solutions have stressed the fact that the sonoluminescence (SL) emissions are the highest ever found, but at the same time the bubble moves in orbits. We have fixed the SL bubble spatially and at the same time we have reached higher SL emissions using another harmonic acoustic signal to produce the acoustic excitation. Multiple harmonic excitation produces up to a fourfold increase in SL emissions, reaching the peak value of about 40 microW for a moving bubble and 15 microW for a nonmoving bubble. The ability to have a bright stationary bubble also opens new research opportunities. In particular, we develop a new method to measure the absolute radius evolution of the bubble that exploits this stability.

Positional stability as the light emission limit in sonoluminescence with sulfuric acid.

We studied a single bubble sonoluminescence system consisting of an argon bubble in a sulfuric acid aq. solution. We experimentally determined the relevant variables of the system. We also measured the bubble position, extent of the bubble orbits, and light intensity as a function of acoustic pressure for different argon concentrations. We find that the Bjerknes force is responsible for the bubble mean position and this imposes a limitation in the maximum acoustic pressure that can be applied to the bubble. The Rayleigh-Taylor instability does not play a role in this system and, at a given gas concentration, the SL intensity depends more on the bubble time of collapse than any other investigated parameter.

Numerical and experimental study of dissociation in an air-water single-bubble sonoluminescence system.

We performed a comprehensive numerical and experimental analysis of dissociation effects in an air bubble in water acoustically levitated in a spherical resonator. Our numerical approach is based on suitable models for the different effects considered. We compared model predictions with experimental results obtained in our laboratory in the whole phase parameter space, for acoustic pressures from the bubble dissolution limit up to bubble extinction. The effects were taken into account simultaneously to consider the transition from nonsonoluminescence to sonoluminescence bubbles. The model includes (1) inside the bubble, transient and spatially nonuniform heat transfer using a collocation points method, dissociation of O2 and N2, and mass diffusion of vapor in the noncondensable gases; (2) at the bubble interface, nonequilibrium evaporation and condensation of water and a temperature jump due to the accommodation coefficient; (3) in the liquid, transient and spatially nonuniform heat transfer using a collocation points method, and mass diffusion of the gas in the liquid. The model is completed with a Rayleigh-Plesset equation with liquid compressible terms and vapor mass transfer. We computed the boundary for the shape instability based on the temporal evolution of the computed radius. The model is valid for an arbitrary number of dissociable gases dissolved in the liquid. We also obtained absolute measurements for R(t) using two photodetectors and Mie scattering calculations. The robust technique used allows the estimation of experimental results of absolute R0 and P(a). The technique is based on identifying the bubble dissolution limit coincident with the parametric instability in (P(a),R0) parameter space. We take advantage of the fact that this point can be determined experimentally with high precision and replicability. We computed the equilibrium concentration of the different gaseous species and water vapor during collapse as a function of P(a) and R0. The model obtains from first principles the result that in sonoluminescence the bubble is practically 100% argon for air dissolved in water. Therefore, the dissociation reactions in air bubbles must be taken into account for quantitative computations of maximum temperatures. The agreement found between the numerical and experimental data is very good in the whole parameter space explored. We do not fit any parameter in the model. We believe that we capture all the relevant physics with the model.

Proposed method to estimate the liquid-vapor accommodation coefficient based on experimental sonoluminescence data.

We used the temporal evolution of the bubble radius in single-bubble sonoluminescence to estimate the water liquid-vapor accommodation coefficient. The rapid changes in the bubble radius that occur during the bubble collapse and rebounds are a function of the actual value of the accommodation coefficient. We selected bubble radius measurements obtained from two different experimental techniques in conjunction with a robust parameter estimation strategy and we obtained that for water at room temperature the mass accommodation coefficient is in the confidence interval [0.217,0.329].