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We report on in vitro wound-healing and cell-growth studies under the influence of radio-frequency (rf) cell stimuli. These stimuli are supplied either by piezoactive surface acoustic waves (SAWs) or by microelectrode-generated electric fields, both at frequencies around 100 MHz. Employing live-cell imaging, we studied the time- and power-dependent healing of artificial wounds on a piezoelectric chip for different cell lines. If the cell stimulation is mediated by piezomechanical SAWs, we observe a pronounced, significant maximum of the cell-growth rate at a specific SAW amplitude, resulting in an increase of the wound-healing speed of up to 135 ± 85% as compared to an internal reference. In contrast, cells being stimulated only by electrical fields of the same magnitude as the ones exposed to SAWs exhibit no significant effect. In this study, we investigate this effect for different wavelengths, amplitude modulation of the applied electrical rf signal, and different wave modes. Furthermore, to obtain insight into the biological response to the stimulus, we also determined both the cell-proliferation rate and the cellular stress levels. While the proliferation rate is significantly increased for a wide power range, cell stress remains low and within the normal range. Our findings demonstrate that SAW-based vibrational cell stimulation bears the potential for an alternative method to conventional ultrasound treatment, overcoming some of its limitations.
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Estimulação Acústica/métodos , Som/efeitos adversos , Vibração/uso terapêutico , Cicatrização/efeitos da radiação , Estimulação Acústica/efeitos adversos , Estimulação Acústica/instrumentação , Animais , Linhagem Celular , Linhagem Celular Tumoral , Movimento Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Terapia Combinada/efeitos adversos , Terapia Combinada/instrumentação , Terapia Combinada/métodos , Cães , Eletrodos , Humanos , Células Madin Darby de Rim Canino , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de OxigênioRESUMO
We report a comprehensive study of the impact of the structural properties in radial GaAs-Al0.3Ga0.7As nanowire-quantum well heterostructures on the optical recombination dynamics and electrical transport properties, emphasizing particularly the role of the commonly observed variations of the quantum well thickness at different facets. Typical thickness fluctuations of the radial quantum well observed by transmission electron microscopy lead to pronounced localization. Our optical data exhibit clear spectral shifts and a multipeak structure of the emission for such asymmetric ring structures resulting from spatially separated, yet interconnected quantum well systems. Charge carrier dynamics induced by a surface acoustic wave are resolved and prove efficient carrier exchange on native, subnanosecond time scales within the heterostructure. Experimental findings are corroborated by theoretical modeling, which unambiguously show that electrons and holes localize on facets where the quantum well is the thickest and that even minute deviations of the perfect hexagonal shape strongly perturb the commonly assumed 6-fold symmetric ground state.
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The separation of deuterium from hydrogen still remains a challenging and industrially relevant task. Compared to traditional cryogenic methods for separation, based on different boiling points of H2 and D2 , the use of ultramicroporous materials offers a more efficient alternative method. Due to their rigid structures, permanently high porosity, tunable pore sizes and adjustable internal surface properties, metal-organic frameworks (MOFs), a class of porous materials built through the coordination between organic linkers and metal ions/clusters, are more suitable for this approach than zeolites or carbon-based materials. Herein, dynamic gas flow studies on H2 /D2 quantum sieving in MFU-4, a metal-organic framework with ultra-narrow pores of 2.5â Å, are presented. A specially designed sensor with a very fast response based on surface acoustic waves is used. On-chip measurements of diffusion rates in the temperature range 27-207â K reveal a quantum sieving effect, with D2 diffusing faster than H2 below 64â K and the opposite selectivity above this temperature. The experimental results obtained are confirmed by molecular dynamic simulation regarding quantum sieving of H2 and D2 on MOFs for which a flexible framework approach was used for the first time.
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BACKGROUND: Rosetting is associated with severe malaria and a primary cause of death in Plasmodium falciparum infections. Detailed understanding of this adhesive phenomenon may enable the development of new therapies interfering with rosette formation. For this, it is crucial to determine parameters such as rosetting and parasitaemia of laboratory strains or patient isolates, a bottleneck in malaria research due to the time consuming and error prone manual analysis of specimens. Here, the automated, free, stand-alone analysis software automated rosetting analyzer for micrographs (ARAM) to determine rosetting rate, rosette size distribution as well as parasitaemia with a convenient graphical user interface is presented. METHODS: Automated rosetting analyzer for micrographs is an executable with two operation modes for automated identification of objects on images. The default mode detects red blood cells and fluorescently labelled parasitized red blood cells by combining an intensity-gradient with a threshold filter. The second mode determines object location and size distribution from a single contrast method. The obtained results are compared with standardized manual analysis. Automated rosetting analyzer for micrographs calculates statistical confidence probabilities for rosetting rate and parasitaemia. RESULTS: Automated rosetting analyzer for micrographs analyses 25 cell objects per second reliably delivering identical results compared to manual analysis. For the first time rosette size distribution is determined in a precise and quantitative manner employing ARAM in combination with established inhibition tests. Additionally ARAM measures the essential observables parasitaemia, rosetting rate and size as well as location of all detected objects and provides confidence intervals for the determined observables. No other existing software solution offers this range of function. The second, non-malaria specific, analysis mode of ARAM offers the functionality to detect arbitrary objects. CONCLUSIONS: Automated rosetting analyzer for micrographs has the capability to push malaria research to a more quantitative and statistically significant level with increased reliability due to operator independence. As an installation file for Windows © 7, 8.1 and 10 is available for free, ARAM offers a novel open and easy-to-use platform for the malaria community to elucidate resetting.
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Interpretação de Imagem Assistida por Computador , Malária Falciparum/diagnóstico por imagem , Parasitemia/diagnóstico por imagem , Plasmodium falciparum/isolamento & purificação , Software , Eritrócitos/fisiologia , Malária Falciparum/sangue , Malária Falciparum/fisiopatologia , Microscopia/instrumentação , Parasitemia/parasitologia , Reprodutibilidade dos Testes , Formação de Roseta/instrumentaçãoRESUMO
A low cost version to enable high temperature MAS NMR experiments at temperatures of up to 700°C and spinning speeds of up to 10kHz is presented. The method relies on inductive heating using a metal coated rotor insert. The metal coating is accomplished via a two step process involving physical vapor deposition and galvanization.
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We probe and control the optical properties of emission centers forming in radial heterostructure GaAs-Al0.3Ga0.7As nanowires and show that these emitters, located in Al0.3Ga0.7As layers, can exhibit quantum-dot like characteristics. We employ a radio frequency surface acoustic wave to dynamically control their emission energy, and occupancy state on a nanosecond time scale. In the spectral oscillations, we identify unambiguous signatures arising from both the mechanical and electrical component of the surface acoustic wave. In addition, different emission lines of a single emission center exhibit pronounced anticorrelated intensity oscillations during the acoustic cycle. These arise from a dynamically triggered carrier extraction out of the emission center to a continuum in the radial heterostructure. Using finite element modeling and Wentzel-Kramers-Brillouin theory we identify quantum tunneling as the underlying mechanism. These simulation results quantitatively reproduce the observed switching and show that in our systems these emission centers are spatially separated from the continuum by >10.5 nm.
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BACKGROUND: Understanding of malaria pathogenesis caused by Plasmodium falciparum has been greatly deepened since the introduction of in vitro culture system, but the lack of a method to enrich ring-stage parasites remains a technical challenge. Here, a novel way to enrich red blood cells containing parasites in the early ring stage is described and demonstrated. METHODS: A simple, straight polydimethylsiloxane microchannel connected to two syringe pumps for sample injection and two height reservoirs for sample collection is used to enrich red blood cells containing parasites in the early ring stage (8-10 h p.i.). The separation is based on the non-inertial hydrodynamic lift effect, a repulsive cell-wall interaction that enables continuous and label-free separation with deformability as intrinsic marker. RESULTS: The possibility to enrich red blood cells containing P. falciparum parasites at ring stage with a throughput of ~12,000 cells per hour and an average enrichment factor of 4.3 ± 0.5 is demonstrated. CONCLUSION: The method allows for the enrichment of red blood cells early after the invasion by P. falciparum parasites continuously and without any need to label the cells. The approach promises new possibilities to increase the sensitivity of downstream analyses like genomic- or diagnostic tests. The device can be produced as a cheap, disposable chip with mass production technologies and works without expensive peripheral equipment. This makes the approach interesting for the development of new devices for field use in resource poor settings and environments, e.g. with the aim to increase the sensitivity of microscope malaria diagnosis.
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Eritrócitos/parasitologia , Citometria de Fluxo/métodos , Malária Falciparum/sangue , Malária Falciparum/parasitologia , Técnicas Analíticas Microfluídicas/métodos , Plasmodium falciparum/isolamento & purificação , Citometria de Fluxo/instrumentação , Humanos , Técnicas Analíticas Microfluídicas/instrumentação , Parasitemia/sangue , Parasitemia/parasitologiaRESUMO
We quantify endocytosis-like nanoparticle (NP) uptake of model membranes as a function of temperature and, therefore, phase state. As model membranes, we use giant unilamellar vesicles (GUV) consisting of 1,2-dipentadecanoyl-sn-glycero-3-phosphocholine (15:0 PC). Time-series micrographs of the vesicle shrinkage show uptake rates that are a highly nonlinear function of temperature. A global maximum appears close to the main structural phase transition at T = Tm + 3 K = 37 °C and a minor peak at the pretransition T = Tp = 22 °C. The quality of linear fits to the shrinkage, and thus uptake kinetics, reveals a deviation from the linear trend at the vesicle shrinkage peaks. Taking values for the bending modulus as a function of temperature from literature and Helfrich's model allows us to draw qualitative conclusions on the membrane tension and the adhesion of the NP to the membrane as a function of temperature. These findings provide valuable insights into the dynamic interplay between temperature, membrane phase transitions, and NP uptake, shedding light on the complex behavior of biological membranes.
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Phase decomposition in lipid membranes has been the subject of numerous investigations by both experiment and theoretical simulation, yet quantitative comparisons of the simulated data to the experimental results are rare. In this work, we present a novel way of comparing the temporal development of liquid-ordered domains obtained from numerically solving the Cahn-Hilliard equation and by inducing a phase transition in giant unilamellar vesicles (GUVs). Quantitative comparison is done by calculating the structure factor of the domain pattern. It turns out that the decomposition takes place in three distinct regimes in both experiment and simulation. These regimes are characterized by different rates of growth of the mean domain diameter, and there is quantitative agreement between experiment and simulation as to the duration of each regime and the absolute rate of growth in each regime.
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Lipídeos de Membrana/química , Microscopia de Fluorescência , Lipossomas Unilamelares/químicaRESUMO
Acousto-electric charge conveyance induced by a surface acoustic wave (SAW) is employed to dissociate photogenerated excitons. Over macroscopic distances, both electrons and holes are injected sequentially into a remotely positioned, isolated and high quality quantum emitter, a self-assembled quantum post. This process is found to be highly efficient and to exhibit improved stability at high acoustic powers when compared to direct optical pumping at the position of the quantum post. These characteristics are attributed to the wide matrix quantum well in which charge conveyance occurs and to the larger number of carriers available for injection in the remote configuration, respectively. The emission of such pumped quantum posts is dominated by recombination of neutral excitons and fully directional when the propagation direction of the SAW and the position of the quantum post are reversed.
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We report on optical experiments performed on individual GaAs nanowires and the manipulation of their temporal emission characteristics using a surface acoustic wave. We find a pronounced, characteristic suppression of the emission intensity for the surface acoustic wave propagation aligned with the axis of the nanowire. Furthermore, we demonstrate that this quenching is dynamical as it shows a pronounced modulation as the local phase of the surface acoustic wave is tuned. These effects are strongly reduced for a surface acoustic wave applied in the direction perpendicular to the axis of the nanowire due to their inherent one-dimensional geometry. We resolve a fully dynamic modulation of the nanowire emission up to 678 MHz not limited by the physical properties of the nanowires.
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Arsenicais/química , Gálio/química , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Acústica , Luz , Teste de Materiais , Tamanho da Partícula , Espalhamento de Radiação , VibraçãoRESUMO
With a view on adding to their use in trace gas sensing, we perform a combined experimental and theoretical study of the change of the conductivity of a metal organic framework (iron (1,2,3)-triazolate, Fe(ta)2) with the uptake of chemically inert gases. To align our first-principles calculations with experimental measurements, we perform an ensemble average over different microscopic arrangements of the gas molecules in the pores of the metal-organic framework (MOF). Up to the experimentally reachable limit of gas uptake, we find a good agreement between both approaches. Thus, we can employ theory to further interpret our experimental results in terms of changes to the parameters of the Bardeen-Shockley band theory, electron-phonon coupling (in the form of the deformation potential), bulk modulus, and carrier effective mass. We find the first of these to be most strongly influenced through the gas uptake. Furthermore, we find the changes to the deformation potential to strongly depend on the individual microscopic arrangements of molecules in the pores of the MOF. This hints at a possible synthetic engineering of the material, e.g., by closing off certain pores, for a stronger, more interpretable electric response upon gas sorption.
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Direct conversion of X-ray irradiation using a semiconductor material is an emerging technology in medical and material sciences. Existing technologies face problems, such as sensitivity or resilience. Here, we describe a novel class of X-ray sensors based on GaN thin film and GaN/AlGaN high-electron-mobility transistors (HEMTs), a promising enabling technology in the modern world of GaN devices for high power, high temperature, high frequency, optoelectronic, and military/space applications. The GaN/AlGaN HEMT-based X-ray sensors offer superior performance, as evidenced by higher sensitivity due to intensification of electrons in the two-dimensional electron gas (2DEG), by ionizing radiation. This increase in detector sensitivity, by a factor of 104 compared to GaN thin film, now offers the opportunity to reduce health risks associated with the steady increase in CT scans in today's medicine, and the associated increase in exposure to harmful ionizing radiation, by introducing GaN/AlGaN sensors into X-ray imaging devices, for the benefit of the patient.
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Spatially addressable arrays of molecules embedded in or anchored to supported lipid bilayers are important for on-chip screening and binding assays; however, methods to sort or accumulate components in a fluid membrane on demand are still limited. Here we apply in-plane surface acoustic shear waves (SAWs) to laterally accumulate double-stranded DNA segments electrostatically bound to a cationic supported lipid bilayer. The fluorescently labeled DNA segments are found to segregate into stripe patterns with a spatial frequency corresponding to the periodicity of the standing SAW wave (~10 µm). The DNA molecules are accumulated 10-fold in the regions of SAW antinodes. The superposition of two orthogonal sets of SAW sources creates checkerboard like arrays of DNA demonstrating the potential to generate arrayed fields dynamically. The pattern relaxation time of 0.58 s, which is independent of the segment length, indicates a sorting and relaxation mechanism dominated by lipid diffusion rather than DNA self-diffusion.
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DNA/metabolismo , Dispositivos Lab-On-A-Chip , Bicamadas Lipídicas/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos/métodos , DNA/química , Difusão , Fluorescência , Bicamadas Lipídicas/química , Análise de Sequência com Séries de Oligonucleotídeos/instrumentação , Som , Eletricidade Estática , Xantenos/análiseRESUMO
Individual self-assembled quantum dots and quantum posts are studied under the influence of a surface acoustic wave. In optical experiments we observe an acoustically induced switching of the occupancy of the nanostructures along with an overall increase of the emission intensity. For quantum posts, switching occurs continuously from predominantly charged excitons (dissimilar number of electrons and holes) to neutral excitons (same number of electrons and holes) and is independent of whether the surface acoustic wave amplitude is increased or decreased. For quantum dots, switching is nonmonotonic and shows a pronounced hysteresis on the amplitude sweep direction. Moreover, emission of positively charged and neutral excitons is observed at high surface acoustic wave amplitudes. These findings are explained by carrier trapping and localization in the thin and disordered two-dimensional wetting layer on top of which quantum dots nucleate. This limitation can be overcome for quantum posts where acoustically induced charge transport is highly efficient in a wide lateral matrix-quantum well.
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Membrane-associated enzymes have been found to behave differently qualitatively and quantitatively in terms of activity. These findings were highly debated in the 1970s and many general correlations and reaction specific models have been proposed, reviewed, and discarded. However, new biological applications brought up the need for clarification and elucidation. To address literature shortcomings, we chose the intrinsically water-soluble enzyme a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) and large unilamellar vesicles with a relative broad phase transition. We here present activity measurements of ADAMTS13 in the freely dissolved state and the membrane associated state for phosphocholine lipids with different acyl-chain lengths (13:0, 14:0 and 15:0) and thus main phase transition temperatures. While the freely dissolved enzyme shows a simple Arrhenius behavior, the activity of membrane associated ADAMTS13 in addition shows a peak. This peak temperature correlates with the main phase transition temperature of the used lipids. These findings support an alternative theory of catalysis. This theory predicts a correlation of the membrane associated activity and the heat capacity, as both are susceptibilities of the same surface Gibb's free energy, since the enzyme is attached to the membrane.
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Proteína ADAMTS13/química , Bicamadas Lipídicas/química , Fosfolipídeos/química , Proteína ADAMTS13/metabolismo , Humanos , Bicamadas Lipídicas/metabolismo , Transição de Fase , Fosfolipídeos/metabolismo , Solubilidade , Temperatura , Água/químicaRESUMO
A Love-wave based biosensor is introduced for analyzing a standardized wound healing assay by observing cell growth and quantifying cell detachment processes. Utilizing the piezoelectric material LiTaO3 36° XY-cut with a thin SiO2-cover layer, shear horizontal surface acoustic waves (SAW) are excited and detected by a set of Interdigital Transducers. Epithelial cells, being cultivated on the substrate and invading the sensors delay line cause a phase shift in the transmitted SAW signal. This phase shift correlates exactly with the surface coverage of the invading cells. After wound healing, emerging fluctuations in the phase shift signal provide information about the cell growth in a confluent cell layer. Additionally, the signal slope allows to quantify the cell detachment process induced by apoptosis, necrosis or cell lysis substances, respectively. Furthermore, culture conditions like temperature or osmolality can be simultaneously monitored by SAW. Based on a theoretical approach and using FEM simulations, we identified the acoustoelectric interaction as the main reason for the phase shift in various frequency- and time-dependent studies. Our model is validated by experimental data and allows predicting the phase change caused by variations in the cell-substrate distance or the volume ratio of the nucleus and the complete cell.
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For Plasmodium falciparum related malaria (B50), one of the outstanding host factors for the development of severe disease is the ABO blood group of malaria patients, where blood group O reduces the probability of severe disease as compared to individuals of groups A, B, or AB. In this report, we investigate the stability of rosette aggregates in malaria caused by Plasmodium falciparum in microflows. These flows are created in microfluidic channels with stenosis-like constrictions of different widths down to ones narrower as the rosette's diameter. High speed videos were recorded and analyzed by a MATLAB© based tracking software (SURF: SUrvival of Rosettes in Flow). We find a correlation of rosette size, channel diameter, and blood group regarding the mobility of the rosettes. Following the concept of a thermodynamic model, we find a critical width of the stenosis for rosette rupture during their passage. Our data reveal that under physiologically relevant conditions, rosettes in blood group A have a higher rosette frequency and stability as compared to blood group O (BG O), which constitutes a crucial factor promoting the observed protection in BG O individuals against severe malaria in non-O individuals.
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We direct the motion of droplets in microfluidic channels using a surface acoustic wave device. This method allows individual drops to be directed along separate microchannel paths at high volume flow rates, which is useful for droplet sorting.
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Superparamagnetic beads in giant unilamellar vesicles are used to facilitate magnetic manipulation, positioning, agitation and mixing of ultrasmall liquid volumes. Vesicles act as leakproof picoliter reaction vessels in an aqueous bulk solution and can be deliberately conveyed by an external magnetic field to a designated position. Upon application of an external magnetic field the beads align to form extended chains. In a rotating magnetic field chains break up into smaller fragments caused by the interplay of viscous friction and magnetic attraction. This process obeys a simple relationship and can be exploited to enhance mixing of the vesicle content and the outer solution or adjacent vesicle volumes exactly at the position of release.