Protein Adsorption at the Air-Water Interface by a Charge Sensing Interferometric Technique.

Protein uptake at the interface of a millimeter-sized air bubble in water is investigated by a recently developed differential interferometric technique. The technique allows the study of capillary waves with amplitudes around 10-9 m, excited at the surface of the bubble by an electric field of intensity on the order of 10 V/cm. When one studies the resonant modes of the bubble (radial and shape modes), it is possible to assess variations of interfacial properties and, in particular, of the net surface charge as a function of bulk protein concentration. Sensing the interfacial charge, the technique enables us to follow the absorption process in conditions of low concentrations, not easily assessable by other methods. We focus on bovine serum albumin (BSA) and lysozyme as representatives of typical globular proteins. To provide comprehensive insight into the novelty of the technique, we also investigated the equilibrium adsorption of sodium dodecyl sulfate (SDS) ionic surfactant for bulk concentrations at hundreds of times lower than the Critical Micelle Concentration (CMC). Results unveil how the absorption of charged molecules affects the amplitudes of the bubble resonant modes even before affecting the frequencies in a transition-like fashion. Different adsorption models are proposed and developed. They are validated against the experimental findings by comparing frequency and amplitude data. By measuring the charging rate of the bubble interface, we have followed the absorption kinetics of BSA and lysozyme recognizing a slow, energy barrier limited phenomena with characteristic times in agreement with data in the literature. The evaluation of the surface excess concentration (Γ) of BSA and SDS at equilibrium is obtained by monitoring charge uptake. At the investigated low bulk concentrations, reliable comparisons with literature data from equilibrium surface tension isotherm models are reported.

Nanometric Surface Oscillation Spectroscopy of Water-Poor Microemulsions.

Selectively exchanging metal complexes between emulsified water-poor microemulsions and concentrated solutions of mixed electrolytes is the core technology for strategic metal recycling. Nanostructuration triggered by solutes present in the organic phase is understood, but little is known about fluctuations of the microemulsion-water interface. We use here a modified version of an optoelectric device initially designed for air bubbles, in order to evidence resonant electrically induced surface waves of an oily droplet suspended in an aqueous phase. Resonant waves of nanometer amplitude of a millimeter-sized microemulsion droplet containing a common ion-specific extractant diluted by dodecane and suspended in a solution of rare earth nitrate are evidenced for the first time with low excitation fields (5 V/cm). From variation of the surface wave spectrum with rare earth concentration, we evidence uptake of rare-earth ions at the interface and at higher concentration the formation of a thin "crust" of liquid crystal forming at unusually low concentration, indicative of a surface induced phase transition. The effect of the liquid crystal structure on the resonance spectrum is backed up by a model, which is used to estimate crust thickness.

Anomalous Behavior of Ultra-Low-Amplitude Capillary Waves. A Glimpse of the Viscoelastic Properties of Interfacial Water?

We investigate, both theoretically and by a differential interferometric technique, the behavior of large-wavelength capillary waves (of the order of 10-4 m) selectively excited at the surface of drops and bubbles with typical eigenfrequencies of the order of 102 Hz. The resonance peaks of gas bubbles or hydrocarbon drops in water (radius less than 1 mm) highlight anomalously small dissipation in the region of ultralow (sub-nanometric) oscillation amplitudes, reaching a plateau at higher amplitudes. This is in sharp contrast to the usual oscillating systems, where an anomalous behavior holds at large amplitudes alone. Dissipation is strongly dependent on the excited vibrational modes and, in spite of remarkable numerical differences, water-vapor and water-hydrocarbon interfaces exhibit the same overall trend. A phenomenological model was developed, based on the assumption that water possesses a threshold viscoelasticity, above which it behaves like a regular viscous fluid. The well-known Deborah number was then estimated within the anomalous region and found to lie in the range of viscoelastic fluids. In agreement with previous studies of nanohydrodynamics (e.g., atomic force microscopy measurements with sub-nanometric tip motions), the present one lends support to the idea that every self-aggregating fluid exhibits yield stress behavior, including classical Newtonian fluids like water. The essential requirement is that the applied perturbation lie below a critical threshold, above which viscous behavior is recovered. Our differential interferometric technique seems particularly suitable for this type of studies, as it allows measurement of long-wavelength capillary waves with sub-nanometric resolution on the oscillation amplitudes.

Oscillations of Bubble Shape Cause Anomalous Surfactant Diffusion: Experiments, Theory, and Simulations.

We investigate, both theoretically and experimentally, the role played by the oscillations of the cell membrane on the capture rate of substances freely diffusing around the cell. To obtain quantitative results, we propose and build up a reproducible and tunable biomimetic experimental model system to simulate the phenomenon of an oscillation-enhanced (or depressed) capture rate (chemoreception) of a diffusant. The main advantage compared to real biological systems is that the different oscillation parameters (type of deformation, frequencies, and amplitudes) can be finely tuned. The model system that we use is an anchored gas drop submitted to a diffusive flow of charged surfactants. When the surfactant meets the surface of the bubble, it is reversibly adsorbed. Bubble oscillations of the order of a few nanometers are selectively excited, and surfactant transport is accurately measured. The surfactant concentration past the oscillating bubbles was detected by conductivity measurements. The results highlight the role of surface oscillations on the diffusant capture rate. Particularly unexpected is the onset of intense overshoots during the adsorption process. The phenomenon is particularly relevant when the bubbles are exposed to intense forced oscillations near resonance.

Trapping of Sodium Dodecyl Sulfate at the Air-Water Interface of Oscillating Bubbles.

We report that at very low initial bulk concentrations, a couple of hundred times below the critical micellar concentration (CMC), anionic surfactant sodium dodecyl sulfate (SDS) adsorbed at the air-water interface of a gas bubble cannot be removed, on the time scale of the experiment (hours), when the surrounding solution is gently replaced by pure water. Extremely sensitive interferometric measurements of the resonance frequency of the bubble-forced oscillations give precise access to the concentration of the surfactant monolayer. The bulk-interface dynamic exchange of SDS molecules is shown to be inhibited below a concentration which we believe refers to a kind of gas-liquid phase transition of the surface monolayer. Above this threshold we recover the expected concentration-dependent desorption. The experimental observations are interpreted within simple energetic considerations supported by molecular dynamics (MD) calculations.

Out of equilibrium divergence of dissipation in an oscillating bubble coated by surfactants.

We report measurements of the relaxation and resonance frequency of forced oscillating bubbles covered by a layer of surface-active molecules, the anionic surfactant sodium dodecyl sulfate (SDS). Less systematic investigations have been also carried out on neutral and cationic surfactants. A divergence of the viscous damping is observed at a very low bulk concentration. Subtle variations in the resonance peak are also measured. Bubble oscillations are driven by an electric field and measured with a sensitive interferometric technique. Results are interpreted with a model which takes care of the coupling between the dynamics of fluid surface oscillations and the properties of a surfactant monolayer in the vicinity of the phase transition from a gas-like distribution to a liquid-like assembly (the so-called gas-LE transition). Important charge effects are also considered. The basic assumptions of the model (cooperative adsorption of the surfactant at the air-water interface and coupling between the shape of the deformed surface and the local surfactant concentration) have been fully confirmed by extensive coarse-grained molecular dynamics simulations on model systems.

New interferometric technique to evaluate the electric charge of gas bubbles in liquids.

We report a new interferometric technique to measure the electric charge at the gas-liquid interface of a bubble in a liquid. The bubble rests by buoyancy against an electrode, and an alternating electric field excites its capillary oscillations. The oscillation amplitude of the quadrupolar mode frequency is measured by the interferometer, and it is used to evaluate the electric charge. The mode frequency scales with the square root of the interfacial tension and with a -(3)/(2) power law as a function of the bubble radius. For bubbles in the millimeter diameter range in pure water, the measured negative charge scales with the square of the radius, hence, giving a constant surface charge density on the order of 1.8 × 10(-5) C m(-2), which is rather consistent with the electrophoretic values reported in the literature.

Structural aspects of ganglioside-containing membranes.

The demand for understanding the physical role of gangliosides in membranes is pressing, due to the high number of diverse and crucial biological functions in which they are involved, needing a unifying thread. To this purpose, model systems including gangliosides have been subject of extensive structural studies. Although showing different levels of complication, all models share the need for simplicity, in order to allow for physico-chemical clarity, so they keep far from the extreme complexity of the true biological systems. Nonetheless, as widely agreed, they provide a basic hint on the structural contribution specific molecules can pay to the complex aggregate. This topic we address in the present review. Gangliosides are likely to play their physical role through metamorphism, cooperativity and demixing, that is, they tend to segregate and identify regions where they can dictate and modulate the geometry and the topology of the structure, and its mechanical properties. Strong three-dimensional organisation and cooperativity are exploited to scale up the local arrangement hierarchically from the nano- to the mesoscale, influencing the overall morphology of the structure.

An interferometric technique to study capillary waves.

We describe a new interferometric technique to study gas-liquid and liquid-liquid interfaces. Bubbles and drops are subjected to an alternating electric field which excites capillary oscillations at the interface, if charged. Bubble or drop deformation is detected by the change of the internal optical path of a laser beam crossing perpendicular to the oscillation axis. Due to the closed geometry, a discrete spectrum of stationary oscillation frequencies (normal modes) is excited. The interferometric nature of the measurement and the resonant nature of the oscillation modes concur in allowing for high sensitivity, in the sub-nanometric region. We present a detailed description of the experimental setup and examples of applications of the technique to the study of both gas-liquid and liquid-liquid interfaces, either naked or with adsorbed surfactant monolayers, for bubbles and drops with diameter~1mm. In particular, the resonance frequencies and the width of the resonance peaks depend on the surface tension and the viscous dampening, respectively. We show that, by this new technique, properties of the interface can be accessed with confidence at the sub-nanometer scale, and surface phenomena, like the monolayer phase transition or the peculiarities of adsorption/desorption processes, can be unraveled in concentration regimes which are too low for existing methods.

Multilevel structuring of ganglioside-containing aggregates: from simple micelles to complex biomimetic membranes.

We revisit the structural investigation we performed over the years on gangliosides, biological amphiphiles typically found in the cell membranes of the nervous system of mammalians. Their molecular features, a large and charged saccharidic headgroup connected to a sticky and extended ceramide double tail, strongly dictate their aggregation properties and place ganglioside aggregates at the borderline between the curved world and the flatland. All along we found that unexpected interesting behaviours were induced by the hierarchical propagation of such extreme monomer properties, from the aggregate scale to the mesoscopic phases. In fact, even small changes in the monomer geometry or hindrance result in dramatic aggregate reshaping, due to collective amplification. Surface packing optimization requires preferential mutual orientation of headgroups, giving rise to trapped solid-disordered configurations. The interplay between interparticle and intraparticle interactions gives rise to unexpected behaviours and counterintuitive phase's landscape. In situ modification of monomer properties, operated by enzymatic digestion of aggregated ganglioside headgroups, either causes collective rearrangement or is overwhelmed by collective trapping, depending on their surface density. This aspect is interesting as gangliosides are not evenly distributed in cell membranes, but only in the outer leaflet, where they p]articipate in rafts, functional microdomains enriched in special lipids including cholesterol. We recently found that ganglioside GM1 forces a preferential distribution of cholesterol, constituting a collective structural pair across the membrane. In summary, ganglioside assemblies, through cooperativity, reach a structural complexity comparable or even bigger and more adaptive than that of a protein.

Cooperative behavior of ganglioside molecules in model systems.

A concise discussion of the role of different geometrical conformational states in the process of self-assembling of gangliosides is given. The report focuses on the effects of the geometrical variations occurring in the head group region of gangliosides as reflected on the geometrical properties of the whole assembly. Collective phenomena happening at the water interfacial region are found to be coupled to the phase transition of the lipid moiety, that is, to the well-known order-disorder conformational transition involving the hydrophobic tails. The possible biological relevance of the head group bistability is envisaged.

Shape fluctuations of large unilamellar lipid vesicles observed by laser light scattering: influence of the small-scale structure.

In the present paper, we apply the dynamic laser light scattering technique to investigate the dependence of the characteristic times of thermally induced shape fluctuation of large unilamellar vesicles (LUVs) on bilayer composition. After addressing single-component LUVs made of two common phospholipids, dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC), we investigate the changes in vesicle shape fluctuation times due to the presence of cholesterol and gangliosides (GM1), added in small amounts. The experimental results show that the addition of a second component, even in small amount, to DMPC vesicles induces a change in membrane fluctuation times. Moreover, in the case of ganglioside addition, also the disposition of GM1 within the bilayer is of importance. Quite unexpectedly, the symmetric or asymmetric disposition of GM1 has opposite effects on bilayer dynamics, the first resulting in a "hardening" and the second in a "softening" of the membrane. Those results support that the small-scale structure of the bilayer is important in determining the overall dynamics of the vesicle. They also suggest that the physiological disposition of GM1 in the outer leaflet of real cells has a significative result in mechanical terms, positively affecting the dynamics of the membrane.