Nonreciprocal Dzyaloshinskii-Moriya Magnetoacoustic Waves.

We study the interaction of surface acoustic waves with spin waves in ultrathin CoFeB/Pt bilayers. Because of the interfacial Dzyaloshinskii-Moriya interaction (DMI), the spin wave dispersion is nondegenerate for oppositely propagating spin waves in CoFeB/Pt. In combination with the additional nonreciprocity of the magnetoacoustic coupling itself, which is independent of the DMI, highly nonreciprocal acoustic wave transmission through the magnetic film is observed. We systematically characterize the magnetoacoustic wave propagation in a thickness series of CoFeB(d)/Pt samples as a function of magnetic field magnitude and direction, and at frequencies up to 7 GHz. We quantitatively model our results to extract the strength of the DMI and magnetoacoustic driving fields.

Correlation of in vitro cell adhesion, local shear flow and cell density.

Investigating cell adhesion behavior on biocompatible surfaces under dynamic flow conditions is not only of scientific interest but also a principal step towards development of new medical implant materials. Driven by the improvement of the measurement technique for microfluidic flow fields (scanning particle image velocimetry, sPIV), a semi-automatic correlation of the local shear velocity and the cell detachment probability became possible. The functionality of customized software entitled 'PIVDAC' (Particle Image Velocimetry De-Adhesion Correlation) is demonstrated on the basis of detachment measurements using standard sand-blasted titanium implant material. A thermodynamic rate model is applied to describe the process of cell adhesion and detachment. A comparison of the model and our experimental findings, especially in a mild regime, where the shear flow does not simply tear away all cells from the substrate, demonstrates, as predicted, an increase of detachment rate with increasing shear force. Finally, we apply the method to compare experimentally obtained detachment rates under identical flow conditions as a function of cell density and find excellent agreement with previously reported model simulations that consider pure geometrical effects. The demonstrated method opens a wide field of applications to study various cell lines on novel substrates or in time dependent flow fields.

Acoustotaxis -in vitro stimulation in a wound healing assay employing surface acoustic waves.

A novel, ultrasound based approach for the dynamic stimulation and promotion of tissue healing processes employing surface acoustic waves (SAW) on a chip is presented for the example of osteoblast-like SaOs-2 cells. In our investigations, we directly irradiate cells with SAW on a SiO2 covered piezoelectric LiNbO3 substrate. Observing the temporal evolution of cell growth and migration and comparing non-irradiated to irradiated areas on the chip, we find that the SAW-treated cells exhibit a significantly increased migration as compared to the control samples. Apart from quantifying our experimental findings on the cell migration stimulation, we also demonstrate the full bio compatibility and bio functionality of our SAW technique by using LDH assays. We safely exclude parasitic side effects such as a SAW related increased substrate temperature or nutrient flow by thoroughly monitoring the temperature and the flow field using infrared microscopy and micro particle image velocimetry. Our results show that the SAW induced dynamic mechanical and electrical stimulation obviously directly promotes the cell growth. We conclude that this stimulation method offers a powerful platform for future medical treatment, e.g. being implemented as a implantable biochip with wireless extra-corporal power supply to treat deeper tissue.

Collective lipid bilayer dynamics excited by surface acoustic waves.

We use standing surface acoustic waves to induce coherent phonons in model lipid multilayers deposited on a piezoelectric surface. Probing the structure by phase-controlled stroboscopic x-ray pulses we find that the internal lipid bilayer electron density profile oscillates in response to the externally driven motion of the lipid film. The structural response to the well-controlled motion is a strong indication that bilayer structure and membrane fluctuations are intrinsically coupled, even though these structural changes are averaged out in equilibrium and time integrating measurements. Here the effects are revealed by a timing scheme with temporal resolution on the picosecond scale in combination with the sub-nm spatial resolution, enabled by high brilliance synchrotron x-ray reflectivity.

Circulating but not immobilized N-deglycosylated von Willebrand factor increases platelet adhesion under flow conditions.

The role of von Willebrand factor (VWF) as a shear stress activated platelet adhesive has been related to a coiled-elongated shape conformation. The forces dominating this transition have been suggested to be controlled by the proteins polymeric architecture. However, the fact that 20% of VWF molecular weight originates from glycan moieties has so far been neglected in these calculations. In this study, we present a systematic experimental investigation on the role of N-glycosylation for VWF mediated platelet adhesion under flow. A microfluidic flow chamber with a stenotic compartment that allows one to mimic various physiological flow conditions was designed for the efficient analysis of the adhesion spectrum. Surprisingly, we found an increase in platelet adhesion with elevated shear rate, both qualitatively and quantitatively fully conserved when N-deglycosylated VWF (N-deg-VWF) instead of VWF was immobilized in the microfluidic channel. This has been demonstrated consistently over four orders of magnitude in shear rate. In contrast, when N-deg-VWF was added to the supernatant, an increase in adhesion rate by a factor of two was detected compared to the addition of wild-type VWF. It appears that once immobilized, the role of glycans is at least modified if not-as found here for the case of adhesion-negated. These findings strengthen the physical impact of the circulating polymer on shear dependent platelet adhesion events. At present, there is no theoretical explanation for an increase in platelet adhesion to VWF in the absence of its N-glycans. However, our data indicate that the effective solubility of the protein and hence its shape or conformation may be altered by the degree of glycosylation and is therefore a good candidate for modifying the forces required to uncoil this biopolymer.

Propagation of 2D pressure pulses in lipid monolayers and its possible implications for biology.

The existence and propagation of acoustic pressure pulses on lipid monolayers at the air-water interface are directly observed by simple mechanical detection. The pulses are excited by small amounts of solvents added to the monolayer. Controlling the state of the lipid interface, we show that the pulses propagate at velocities c following the lateral compressibility κ. This is manifested by a pronounced minimum in c (∼0.3 m/s) within the transition regime. The role of interface density pulses in biology is discussed, in particular, in the context of communicating localized alterations in protein function (signaling) and nerve pulse propagation.

Chemical and mechanical impact of silica nanoparticles on the phase transition behavior of phospholipid membranes in theory and experiment.

The interaction of nanoparticles (NPs) with lipid membranes is an integral step in the interaction of NPs and living cells. During particle uptake, the membrane has to bend. Due to the nature of their phase diagram, the modulus of compression of these membranes can vary by more than one order of magnitude, and thus both the thermodynamic and mechanical aspects of the membrane have to be considered simultaneously. We demonstrate that silica NPs have at least two independent effects on the phase transition of phospholipid membranes: 1), a chemical effect resulting from the finite instability of the NPs in water; and 2), a mechanical effect that originates from a bending of the lipid membrane around the NPs. Here, we report on recent experiments that allowed us to clearly distinguish both effects, and present a thermodynamic model that includes the elastic energy of the membranes and correctly predicts our findings both quantitatively and qualitatively.

Simultaneously propagating voltage and pressure pulses in lipid monolayers of pork brain and synthetic lipids.

Hydrated interfaces are ubiquitous in biology and appear on all length scales from ions and individual molecules to membranes and cellular networks. In vivo, they comprise a high degree of self-organization and complex entanglement, which limits their experimental accessibility by smearing out the individual phenomenology. The Langmuir technique, however, allows the examination of defined interfaces, the controllable thermodynamic state of which enables one to explore the proper state diagrams. Here we demonstrate that voltage and pressure pulses simultaneously propagate along monolayers comprised of either native pork brain or synthetic lipids. The excitation of pulses is conducted by the application of small droplets of acetic acid and monitored subsequently employing time-resolved Wilhelmy plate and Kelvin probe measurements. The isothermal state diagrams of the monolayers for both lateral pressure and surface potential are experimentally recorded, enabling us to predict dynamic voltage pulse amplitudes of 0.1-3 mV based on the assumption of static mechanoelectrical coupling. We show that the underlying physics for such propagating pulses is the same for synthetic and natural extracted (pork brain) lipids and that the measured propagation velocities and pulse amplitudes depend on the compressibility of the interface. Given the ubiquitous presence of hydrated interfaces in biology, our experimental findings seem to support a fundamentally new mechanism for the propagation of signals and communication pathways in biology (signaling), which is based neither on protein-protein or receptor-ligand interaction nor diffusion.

Thermomechanic-electrical coupling in phospholipid monolayers near the critical point.

Lipid monolayers have been shown to represent a powerful tool in studying mechanical and thermodynamic properties of lipid membranes as well as their interaction with proteins. Using Einstein's theory of fluctuations we here demonstrate that an experimentally derived linear relationship both between transition entropy S and area A as well as between transition entropy and charge q implies a linear relationships between compressibility κT, heat capacity cπ, thermal expansion coefficient αT, and electric capacity CT. We demonstrate that these couplings have strong predictive power as they allow calculating electrical and thermal properties from mechanical measurements. The precision of the prediction increases as the critical point TC is approached.

Acoustic driven flow and lattice Boltzmann simulations to study cell adhesion in biofunctionalized mu-fluidic channels with complex geometry.

Accurately mimicking the complexity of microvascular systems calls for a technology which can accommodate particularly small sample volumes while retaining a large degree of freedom in channel geometry and keeping the price considerably low to allow for high throughput experiments. Here, we demonstrate that the use of surface acoustic wave driven microfluidics systems successfully allows the study of the interrelation between melanoma cell adhesion, the matrix protein collagen type I, the blood clotting factor von Willebrand factor (vWF), and microfluidic channel geometry. The versatility of the tool presented enables us to examine cell adhesion under flow in straight and bifurcated microfluidic channels in the presence of different protein coatings. We show that the addition of vWF tremendously increases (up to tenfold) the adhesion of melanoma cells even under fairly low shear flow conditions. This effect is altered in the presence of bifurcated channels demonstrating the importance of an elaborate hydrodynamic analysis to differentiate between physical and biological effects. Therefore, computer simulations have been performed along with the experiments to reveal the entire flow profile in the channel. We conclude that a combination of theory and experiment will lead to a consistent explanation of cell adhesion, and will optimize the potential of microfluidic experiments to further unravel the relation between blood clotting factors, cell adhesion molecules, cancer cell spreading, and the hydrodynamic conditions in our microcirculatory system.

Transport, separation, and accumulation of proteins on supported lipid bilayers.

Transport, separation, and accumulation of proteins in their natural environment are central goals in protein biotechnology. Miniaturized assays of supported lipid bilayers (SLBs) have been proposed as promising candidates to realize such technology on a chip, but a modular system for the controlled transport of membrane proteins does not exist. In this letter, we demonstrate that standing surface acoustic waves drive the in-plane redistribution of proteins on planar SLBs over macroscopic distances (3.5 mm). Accumulation of proteins in periodic patterns of about 10-fold protein concentration difference is accomplished and shown to relax into the homogeneous state by diffusion. Different proteins separate in individual fractions from a homogeneous distribution and are transported and accumulated into clusters using beats. The modular planar setup has the potential of integrating other lab-on-a-chip tools, for monitoring the membrane-protein integrity or adding microfluidic features for blood screening or DNA analysis.

Surface acoustic wave actuated cell sorting (SAWACS).

We describe a novel microfluidic cell sorter which operates in continuous flow at high sorting rates. The device is based on a surface acoustic wave cell-sorting scheme and combines many advantages of fluorescence activated cell sorting (FACS) and fluorescence activated droplet sorting (FADS) in microfluidic channels. It is fully integrated on a PDMS device, and allows fast electronic control of cell diversion. We direct cells by acoustic streaming excited by a surface acoustic wave which deflects the fluid independently of the contrast in material properties of deflected objects and the continuous phase; thus the device underlying principle works without additional enhancement of the sorting by prior labelling of the cells with responsive markers such as magnetic or polarizable beads. Single cells are sorted directly from bulk media at rates as fast as several kHz without prior encapsulation into liquid droplet compartments as in traditional FACS. We have successfully directed HaCaT cells (human keratinocytes), fibroblasts from mice and MV3 melanoma cells. The low shear forces of this sorting method ensure that cells survive after sorting.

Regulation of adhesion behavior of murine macrophage using supported lipid membranes displaying tunable mannose domains.

Highly uniform, strongly correlated domains of synthetically designed lipids can be incorporated into supported lipid membranes. The systematic characterization of membranes displaying a variety of domains revealed that the equilibrium size of domains significantly depends on the length of fluorocarbon chains, which can be quantitatively interpreted within the framework of an equivalent dipole model. A mono-dispersive, narrow size distribution of the domains enables us to treat the inter-domain correlations as two-dimensional colloidal crystallization and calculate the potentials of mean force. The obtained results demonstrated that both size and inter-domain correlation can precisely be controlled by the molecular structures. By coupling α-D-mannose to lipid head groups, we studied the adhesion behavior of the murine macrophage (J774A.1) on supported membranes. Specific adhesion and spreading of macrophages showed a clear dependence on the density of functional lipids. The obtained results suggest that such synthetic lipid domains can be used as a defined platform to study how cells sense the size and distribution of functional molecules during adhesion and spreading.

Wave propagation in lipid monolayers.

Sound waves are excited on lipid monolayers using a set of planar electrodes aligned in parallel with the excitable medium. By measuring the frequency-dependent change in the lateral pressure, we are able to extract the sound velocity for the entire monolayer phase diagram. We demonstrate that this velocity can also be directly derived from the lipid monolayer compressibility, and consequently displays a minimum in the phase transition regime. This minimum decreases from v(0) = 170 m/s for one-component lipid monolayers down to v(m) = 50 m/s for lipid mixtures. No significant attenuation can be detected confirming an adiabatic phenomenon. Finally, our data propose a relative lateral density oscillation of Deltarho/rho approximately 2%, implying a change in all area-dependent physical properties. Order-of-magnitude estimates from static couplings therefore predict propagating changes in surface potential of 1-50 mV, 1 unit in pH (electrochemical potential), and 0.01 K in temperature, and fall within the same order of magnitude as physical changes measured during nerve pulse propagation. These results therefore strongly support the idea of propagating adiabatic sound waves along nerves as first thoroughly described by Kaufmann in 1989 and recently by Heimburg and Jackson, but already claimed by Wilke in 1912.

Phase-state dependent current fluctuations in pure lipid membranes.

Current fluctuations in pure lipid membranes have been shown to occur under the influence of transmembrane electric fields (electroporation) as well as a result from structural rearrangements of the lipid bilayer during phase transition (soft perforation). We demonstrate that the ion permeability during lipid phase transition exhibits the same qualitative temperature dependence as the macroscopic heat capacity of a D15PC/DOPC vesicle suspension. Microscopic current fluctuations show distinct characteristics for each individual phase state. Although current fluctuations in the fluid phase show spikelike behavior of short timescales (approximately 2 ms) with a narrow amplitude distribution, the current fluctuations during lipid phase transition appear in distinct steps with timescales of approximately 20 ms. We propose a theoretical explanation for the origin of timescales and permeability based on a linear relationship between lipid membrane susceptibilities and relaxation times near the phase transition.

Thermodynamic relaxation drives expulsion in giant unilamellar vesicles.

We investigated the thermodynamic relaxation of giant unilamellar vesicles (GUVs) which contained small vesicles within their interior. Quenching these vesicles from their fluid phase (T > T(m)) through the phase transition in the gel state (T < T(m)) drives the inner vesicles to be expelled from the larger mother vesicle via the accompanying decrease in the vesicle area by approximately 25% which forces a pore to open in the mother vesicle. We demonstrate that the proceeding time evolution of the resulting efflux follows the relaxation of the membrane area and describes the entire relaxation process using an Onsager-like non-equilibrium thermodynamics ansatz. As a consequence of the volume efflux, internal vesicles are expelled from the mother vesicle. Although complete sealing of the pore may occur during the expulsion, the global relaxation dynamics is conserved. Finally, comparison of these results to morphological relaxation phenomena found in earlier studies reveals a universal relaxation behaviour in GUVs. When quenched from the fluid to gel phase the typical time scale of relaxation shows little variation and ranges between 4 and 5 s.

Determination of the conversion factor for silymarin - a solution for the problems associated with new assay methods in the pharmacopoeia.

In pharmacopoeial monographs for herbal drugs and herbal preparations, conventional assay methods such as colorimetry or spectophotometric assays are often replaced by modern, more specific and reliable methods, e.g. liquid chromatography (LC). However, existing dosage recommendations in the monographs on efficacy and safety of herbal medicinal products which are an important basis for licensing procedures do not refer to the mandatory new methods but to the existing photometric methods. The laboratory comparison of the determination of silymarin of Milk Thistle extract shows that a conversion factor can be calculated which allows a correlation between the new and the existing method. It is suggested that this factor should be included in the Ph. Eur. monograph on Milk Thistle extract, allowing reference to dosages given in official monographs (e.g. ESCOP, HMPC). The solution to use a conversion factor should also be applied to other herbal drugs and herbal preparations, especially for standardised extracts.

Change of the effective dimensionality of an Nb/CuNi bilayer in an external magnetic field.

A dimensional crossover of superconducting fluctuations in an external magnetic field, applied parallel to the layers, has been found for superconductor/ferromagnet bilayers of Nb/Cu(41)Ni(59). By lowering the temperature, a reduction of the superconducting nuclei size occurs. As soon as the size of the nuclei becomes smaller than the thickness of the superconducting bilayer structure, the dimensionality changes. The temperature dependence of the fluctuation conductivity exhibits a 2D behaviour in zero and weak magnetic fields in the vicinity of the critical temperature, switching to a 3D behaviour in a strong magnetic field at low temperatures.

Relaxation of ultralarge VWF bundles in a microfluidic-AFM hybrid reactor.

The crucial role of the biopolymer "Von Willebrand factor" (VWF) in blood platelet binding is tightly regulated by the shear forces to which the protein is exposed in the blood flow. Under high-shear conditions, VWFs ability to immobilize blood platelets is strongly increased due to a change in conformation which at sufficient concentration is accompanied by the formation of ultra large VWF bundles (ULVWF). However, little is known about the dynamic and mechanical properties of such bundles. Combining a surface acoustic wave (SAW) based microfluidic reactor with an atomic force microscope (AFM) we were able to study the relaxation of stretched VWF bundles formed by hydrodynamic stress. We found that the dynamical response of the network is well characterized by stretched exponentials, indicating that the relaxation process proceeds through hopping events between a multitude of minima. This finding is in accordance with current ideas of VWF self-association. The longest relaxation time does not show a clear dependence on the length of the bundle, and is dominated by the internal conformations and effective friction within the bundle.

Theoretical and experimental investigations of Coulomb blockade in coupled quantum dot systems.

Two strongly coupled quantum dots are theoretically and experimentally investigated. In conductance measurements on a GaAs based low-dimensional system additional features to the Coulomb blockade have been detected at low temperatures. These regions of finite conductivity are compared with theoretical investigations of a strongly coupled quantum dot system and good agreement between the theoretical and the experimental results has been found.