Sensing lipid bilayer formation and expansion with a microfabricated cantilever array

We show that cantilever array sensors can sense the formation of supported phospholipid bilayers on their surface and that they can monitor changes in mechanical properties of lipid bilayers. Supported lipid bilayers were formed on top of microfabricated cantilevers by vesicle fusion. The formation of bilayers led to a bending of the cantilevers of 70-590 nm comparable to a surface stress of 27-224 mN/m. Physisorption of bilayers of DOPC and other bilayers on the silicon oxide surface of cantilevers led to a tensile bending of about 70 nm whereas formation of chemisorbed bilayers of mixed thiolated (DPPTE) and non-thiolated lipids (DOPC) on the gold side of cantilevers led to a compressive bending of nearly 600 nm which depended on the ratio of DPPTE to DOPC. First results on bending of bilayer-covered cantilevers due to their interaction with the pore-forming peptide melittin are shown. The results demonstrate that cantilever sensors with immobilized bilayers can be used as model systems to investigate mechanical properties of cellular membranes and may be used for screening of membrane processes involving modification, lateral expansion, or contraction of membranes.     

Evaporation of pure liquid sessile and spherical suspended drops: a review

A sessile drop is an isolated drop which has been deposited on a solid substrate where the wetted area is limited by a contact line and characterized by contact angle, contact radius and drop height. Diffusion-controlled evaporation of a sessile drop in an ambient gas is an important topic of interest because it plays a crucial role in many scientific applications such as controlling the deposition of particles on solid surfaces, in ink-jet printing, spraying of pesticides, micro/nano material fabrication, thin film coatings, biochemical assays, drop wise cooling, deposition of DNA/RNA micro-arrays, and manufacture of novel optical and electronic materials in the last decades. This paper presents a review of the published articles for a period of approximately 120 years related to the evaporation of both sessile drops and nearly spherical droplets suspended from thin fibers. After presenting a brief history of the subject, we discuss the basic theory comprising evaporation of micrometer and millimeter sized spherical drops, self cooling on the drop surface and evaporation rate of sessile drops on solids. The effects of drop cooling, resultant lateral evaporative flux and Marangoni flows on evaporation rate are also discussed. This review also has some special topics such as drop evaporation on superhydrophobic surfaces, determination of the receding contact angle from drop evaporation, substrate thermal conductivity effect on drop evaporation and the rate evaporation of water in liquid marbles.     

'Living cantilever arrays' for characterization of mass of single live cells in fluids

The size of a cell is a fundamental physiological property and is closely regulated by various environmental and genetic factors. Optical or confocal microscopy can be used to measure the dimensions of adherent cells, and Coulter counter or flow cytometry (forward scattering light intensity) can be used to estimate the volume of single cells in a flow. Although these methods could be used to obtain the mass of single live cells, no method suitable for directly measuring the mass of single adherent cells without detaching them from the surface is currently available. We report the design, fabrication, and testing of 'living cantilever arrays', an approach to measure the mass of single adherent live cells in fluid using silicon cantilever mass sensor. HeLa cells were injected into microfluidic channels with a linear array of functionalized silicon cantilevers and the cells were subsequently captured on the cantilevers with positive dielectrophoresis. The captured cells were then cultured on the cantilevers in a microfluidic environment and the resonant frequencies of the cantilevers were measured. The mass of a single HeLa cell was extracted from the resonance frequency shift of the cantilever and was found to be close to the mass value calculated from the cell density from the literature and the cell volume obtained from confocal microscopy. This approach can provide a new method for mass measurement of a single adherent cell in its physiological condition in a non-invasive manner, as well as optical observations of the same cell. We believe this technology would be very valuable for single cell time-course studies of adherent live cells.     

High sensitivity of diamond resonant microcantilevers for direct detection in liquids as probed by molecular electrostatic surface interactions

Resonant microcantilevers have demonstrated that they can play an important role in the detection of chemical and biological agents. Molecular interactions with target species on the mechanical microtransducers surface generally induce a change of the beam's bending stiffness, resulting in a shift of the resonance frequency. In most biochemical sensor applications, cantilevers must operate in liquid, even though damping deteriorates the vibrational performances of the transducers. Here we focus on diamond-based microcantilevers since their transducing properties surpass those of other materials. In fact, among a wide range of remarkable features, diamond possesses exceptional mechanical properties enabling the fabrication of cantilever beams with higher resonant frequencies and Q-factors than when made from other conventional materials. Therefore, they appear as one of the top-ranked materials for designing cantilevers operating in liquid media. In this study, we evaluate the resonator sensitivity performances of our diamond microcantilevers using grafted carboxylated alkyl chains as a tool to investigate the subtle changes of surface stiffness as induced by electrostatic interactions. Here, caproic acid was immobilized on the hydrogen-terminated surface of resonant polycrystalline diamond cantilevers using a novel one-step grafting technique that could be also adapted to several other functionalizations. By varying the pH of the solution one could tune the -COO(-)/-COOH ratio of carboxylic acid moieties immobilized on the surface, thus enabling fine variations of the surface stress. We were able to probe the cantilevers resonance frequency evolution and correlate it with the ratio of -COO(-)/-COOH terminations on the functionalized diamond surface and consequently the evolution of the electrostatic potential over the cantilever surface. The approach successfully enabled one to probe variations in cantilevers bending stiffness from several tens to hundreds of millinewtons/meter, thus opening the way for diamond microcantilevers to direct sensing applications in liquids. The evolution of the diamond surface chemistry was also investigated using X-ray photoelectron spectroscopy.     

Contact line dynamics in the late-stage coalescence of diethylene glycol drops

We investigated the contact line dynamics of a composite drop formed as a result of the coalescence during the condensation of two diethylene glycol (DEG) drops at -4 degrees C on a silicon surface. The composite drop relaxes exponentially toward equilibrium with a typical relaxation time, tc, which depends on the equilibrium radius, R, of the composite drop. The value of tc is found to be in the range of 10-100 s for R approximately 1-4 microm. The relaxation dynamics is found to be larger by 6 orders of magnitude than that predicted by bulk hydrodynamics because of high dissipation in the contact line vicinity. Similar to low viscous liquids (water), this high dissipation can be attributed to an Arrhenius factor resulting from the phase change in the contact line vicinity and to the influence of surface defects that pin the contact line.     

Characterisation of alkyl amines at the water/air surface with the drop and bubble profile analysis tensiometry

Pendant drop and buoyant bubble methods have been used to study the surface characteristics of alkyl amines at the water/air surface. The investigated alkyl amines, triethylamine and octylamine, showed unusual changes in the surface tension as a function of time: an initially steep drop and a subsequent steady increase in the surface tension until a value close to the one of the pure water/air system was observed. This phenomenon is explained by the evaporation of the alkyl amines, for which several sets of experiments have been conducted with the pendant drop and buoyant bubble methods. Using an appropriate experimental protocol, the equilibrium adsorption behaviour of the two amines can be quantitatively measured.     

Water evaporation on highly viscoelastic polymer surfaces

Results are reported for a study on the evaporation of water droplets from a highly viscoelastic acrylic polymer surface. These are contrasted with those collected for the same measurements carried out on polydimethylsiloxane (PDMS). For PDMS, the evaporation process involves the expected multistep process including constant drop area, constant contact angle, and finally a combination of these steps until the liquid is gone. In contrast, water evaporation from the acrylic polymer shows a constant drop area mode throughout. Furthermore, during the evaporation process, the drop area actually expands on the acrylic polymer. The single mode evaporation process is consistent with formation of wetting structures, which cannot be propagated by the capillary forces. Expansion of the drop area is attributed to the influence of the drop capillary pressure. Furthermore, the rate of drop area expansion is shown to be dependent on the thickness of the polymer film.     

Static deflection measurements of cantilever arrays reveal polymer film expansion and contraction

An optical static method of detection is used to interpret surface stress induced bending related to cantilevers coated on one side with poly(vinyl alcohol), poly(vinyl butyral-co-vinyl alcohol-co-vinyl acetate), and poly(vinyl chloride-co-vinyl acetate-co-2-hydroxypropyl acrylate), or respectively, PVA, PVB, and PVC, and exposed to various solvent vapors. Results indicate that the adsorption and surface interactions of the different solvent vapors that cause polymer swelling and shrinking lead to rearrangements, which have been shown to change the elastic properties of the polymer film, and subsequently, the spring constant of the polymer coated cantilever. Static deflection measurements allow the direction of cantilever bending to be determined, which adds a new dimension of usefulness for surface functionalized cantilevers as transducers in the development of novel microelectromechanical systems (MEMS).     

Polymer microcantilever biochemical sensors with integrated polymer composites for electrical detection

Micro fabricated sensors based on nanomechanical motion with piezoresistive electrical readout have become a promising biochemical sensing tool. The conventional microcantilever materials are mostly silicon-based. The sensitivity of the sensor depends on Young's modulus of the structural material, thickness of the cantilever as well as on the gauge factor of the piezoresistor. UV patternable polymers such as SU-8 have a very low Young's modulus compared to the silicon-based materials. Polymer cantilevers with a piezoresistive material having a large gauge factor and a lower Young's modulus are therefore highly suited for sensing applications. In this work, a spin coatable and photopatternable mixture of carbon black (CB) and SU-8, with proper dispersion characteristics, has been demonstrated as a piezoresistive thin film for polymer microcantilevers. Results on percolation experiments of SU-8/CB composite and fabrication of piezoresistive SU-8 microcantilevers using this composite are presented. With our controlled dispersion experiments, we could get a uniform piezoresistive thin film of thickness less than 1.2 μm and resistivity of 2.7 Ω cm using 10 wt% of CB in SU-8. The overall thickness of the SU-8/composite/SU-8 is approximately 3 μm. We further present results on the electromechanical characterization and biofunctionalization of the cantilever structures for biochemical sensing applications. These cantilevers show a deflection sensitivity of 0.55 ppm/nm. Since the surface stress sensitivity is 4.1 × 10⁻³ [N/m]⁻¹, these cantilevers can well be used for detection of protein markers for pathological applications.     

High internal phase emulsions: catastrophic phase inversion, stability, and triggered destabilization

We have investigated the formation, drop sizes, and stability of emulsions prepared by hand shaking in a closed vessel in which the emulsion is in contact with a single type of surface during its formation. The emulsions undergo catastrophic phase inversion from oil-in-water (o/w) to water-in-oil (w/o) as the oil volume fraction is increased. We find that the oil volume fraction required for catastrophic inversion exhibits a linear correlation with the oil-water-solid surface contact angle. W/o high internal phase emulsions (HIPEs) prepared in this way contain water drops of diameters in the range 10-100 μm; emulsion drop size depends on the surfactant concentration and method of preparation. W/o HIPEs with large water drops show water separation but w/o HIPEs with small water drops are stable with respect to water separation for more than 100 days. The destabilization of the w/o HIPEs can be triggered by either evaporation of the oil continuous phase or by contact the emulsion with a solid surface of the "wrong" wettability.     

Triple-line behavior and wettability controlled by nanocoated substrates: influence on sessile drop evaporation

In this article, we investigate the influence of the surface properties of substrates on the evaporation process. Using various nanocoatings, it is possible to modify the surface properties of substrates, such as the roughness and the surface energy, while maintaining constant thermal properties. Experiments are conducted under atmospheric conditions with five fluids (methanol, ethanol, propanol, toluene and water) and four coatings (PFC, PTFE, SiOC, and SiO(x)). The various combinations of these fluids and coatings allow for a wide range of drop evaporation properties to be studied: the dynamics of the triple line, the volatility of fluids, and a large range of wettabilities (from 17 to 135°). The experimental data are in very good quantitative agreement with existing models of quasi-steady, diffusion-driven evaporation. The experimental results show that the dynamics of the evaporative rate are proportional to the dynamics of the wetting radius. Thus, the models succeed in describing the evaporative dynamics throughout the evaporation process regardless of the behavior of the triple line. Moreover, the use of various liquids reveals the validity of the models regardless of their volatility. The results also confirm the recent finding of a universal relation for the time evolution of the drop mass, independent of the drop size and initial contact angle. Finally, this study highlights the separate and coupled roles of the triple line and the wettability on the sessile drop evaporation process. Data reveal that the more wet and pinned a drop, the shorter the evaporation time.     

Advanced tip design for liquid phase vibration mode atomic force microscopy

We have fabricated polymer tips for atomic force microscopy in order to elucidate the effects of tip length and shape on cantilever vibration damping in liquids. The vibration damping is investigated by measuring the vibration amplitude of cantilevers as a function of tip-sample distance. The cantilever with a short tip provides a higher damping effect over long tip-sample distances. When the vibration amplitude was rescaled to show the effect of the cantilever width on oscillation damping, the vibration amplitude of cantilevers with various tip lengths was similarly obtained in a long distance range over 50 μm. This similarity is explained by an acoustic damping model in which an acoustic wave is generated by the cantilever. Finally, the results indicate a cantilever with a sufficiently long tip compared to the cantilever width can dramatically reduce the long-range damping effect in a liquid environment.     

Endonuclease-based logic gates and sensors using magnetic force-amplified readout of DNA scission on cantilevers

The endonuclease scission of magnetic particles functionalized with sequence-specific DNAs, which are associated on cantilevers, is followed by the magnetic force-amplified readout of the reactions by the nano-mechanical deflection/retraction of the cantilevers. The systems are employed to develop AND or OR logic gates and to detect single base mismatch specificity of the endonucleases. The two endonucleases EcoRI (E(A)) and AscI (E(B)) are used as inputs. The removal of magnetic particles linked to the cantilever by the duplexes 1/1a and 2/2a via the simultaneous cleavage of the DNAs by E(A) and E(B) leads to the retraction of the magnetically deflected cantilever and to the establishment of the "AND" gate. The removal of the magnetic particles linked to the cantilevers by the duplex 3/3a by either E(A) or E(B) leads to the retraction of the magnetically deflected cantilever and to the establishment of the "OR" gate. The magnetic force-amplified readout of endonuclease activities is also employed to reveal single base mismatch specificity of the biocatalysts.     

Spreading, evaporation, and contact line dynamics of surfactant-laden microdrops

An optical technique based on the reflectivity measurements of a thin film was used to experimentally study the spreading, evaporation, contact line motion, and thin film characteristics of drops consisting of a water-surfactant (polyalkyleneoxide-modified heptamethyltrisiloxane, called superspreader) solution on a fused silica surface. On the basis of the experimental observations, we concluded that the surfactant adsorbs primarily at the solid-liquid and liquid-vapor interfaces near the contact line region. At equilibrium, the completely wetting corner meniscus was associated with a flat adsorbed film having a thickness of approximately 31 nm. The calculated Hamaker constant, A = -4.47 x 10(-)(20) J, shows that this thin film was stable under equilibrium conditions. During a subsequent evaporation/condensation phase-change process, the thin film of the surfactant solution was unstable, and it broke into microdrops having a finite contact angle. The thickness of the adsorbed film associated with the drops was lower than that of the equilibrium meniscus. The drop profiles were experimentally measured and analyzed during the phase-change process as the contact line advanced and receded. The apparent contact angle, the maximum concave curvature near the contact line region, and the convex curvature of the drop increased as the drop grew during condensation, whereas these quantities decreased during evaporation. The position of the maximum concave curvature of the drop moved toward the center of the drop during condensation, whereas it moved away from the center during evaporation. The contact line velocity was correlated to the observed experimental results and was compared with the results of the drops of a pure alcohol. The experimentally obtained thickness profiles, contact angle profiles, and curvature profiles of the drops explain how the surfactant adsorption affects the contact line motion. We found that there was an abrupt change in the velocity of the contact line when the adsorbed film of the surfactant solution was just hydrated or desiccated during the phase-change processes. This result shows the effect of vesicles and aggregates of the surfactant on the shape evolution of the drops. For these surfactant-laden water drops, we found that the apparent contact angle increased during condensation and decreased during evaporation. However, for the drop of a pure liquid (n-butanol and 2-propanol) the apparent contact angle remained constant at a constant velocity during condensation and evaporation. The contact line was pinned during the evaporation and spreading of the surfactant-laden water drops, but it was not pinned for a drop of a pure alcohol (self-similar shape evolution).     

Optofluidic encapsulation of crystalline colloidal arrays into spherical membrane

Double emulsion droplets encapsulating crystalline colloidal arrays (CCAs) with a narrow size distribution were produced using an optofluidic device. The shell phase of the double emulsion was a photocurable resin that was photopolymerized downstream of the fluidic channel within 1 s after drop generation. The present optofluidic synthesis scheme was very effective for fabricating highly monodisperse spherical CCAs that were made structurally stable by in situ photopolymerization of the encapsulating shells. The shell thickness and the number of core emulsion drops could be controlled by varying the flow rates of the three coflowing streams in the dripping regime. The spherical CCAs confined in the shell exhibited distinct diffraction patterns in the visible range, in contrast to conventional film-type CCAs. As a result of their structure, the spherical CCAs exhibited photonic band gaps for normal incident light independent of the position on the spherical surface. This property was induced by heterogeneous nucleation at the smooth wall of the spherical emulsion drop during crystallization into a face-centered cubic (fcc) structure. On the other hand, the solidified shells did not permit the penetration of ionic species, enabling the CCAs to maintain their structure in a continuous aqueous phase of high ionic strength for at least 1 month. In addition, the evaporation of water molecules inside the shell was slowed considerably when the core-shell microparticles were exposed to air: It took approximately 6 h for a suspension encapsulated in a thick shell to evaporate completely, which is approximately 1000 times longer than the evaporation time for water droplets with the same volume. Finally, the spherical CCAs additionally exhibited enhanced stability against external electric fields. The spherical geometry and high dielectric constant of the suspension contributed to reducing the electric field inside the shell, thereby inhibiting the electrophoretic movement of the charged particles.     

Studies of the rate of water evaporation through adsorption layers using drop shape analysis tensiometry

With modified measuring procedure and measuring cell design in the drop profile tensiometer PAT, it became possible to study the rate of water evaporation through adsorbed or spread surface layers. This method was employed to measure the rate of water evaporation from drops covered by adsorbed layers of some proteins and surfactants, in particular n-dodecanol. It was shown that the formation of dense (double or condensed) adsorbed layers of protein and the formation of 2D-condensed n-dodecanol layer decrease the water evaporation rate by 20-25% as compared with pure water. At the same time, the adsorbed layers of ordinary surfactants (sodium dodecyl sulfate and nonionic ethoxylated surfactant C(14)EO(8)) do not affect the water evaporation rate remarkably.     

Nanomechanics of silicon surfaces with atomic force microscopy: an insight to the first stages of plastic deformation

The use of stiff cantilevers with diamond tips allows us to perform nanoindentations on hard covalent materials such as silicon with atomic force microscopy. Thanks to the high sensitivity in the force measurements together with the high resolution upon imaging the surface, we can study nanomechanical properties. At this scale, the surface deforms, following a simple non-Hertzian spring model. The plastic onset can be assessed from a discontinuity in the force-distance curves. Hardness measurements with penetration depths as small as 1 nm yield H= approximately 25 GPa, thus showing a drastic increase with penetration depths below 5 nm.     

Atomic force microscopy measurements of peptide-wrapped single-walled carbon nanotube diameters.

With a vertical resolution of 0.1 nm, atomic force microscopy (AFM) height measurements can be used to determine accurately the diameter of single-walled carbon nanotubes (SWNT) with the assumption that they have circular cross sections. The aim of this article is to draw attention to the need to optimize operating parameters in tapping mode for quantitative AFM height (diameter) analysis of SWNTs. Using silicon tip/cantilever assemblies with force constants ranging from 0.9 to 40 N m(-1), we examined the effect of applied force on the apparent diameter of SWNT wrapped with a 29-residue amphiphilic alpha-helical peptide. A decrease in apparent height (SWNT diameter) with increasing applied force was observed for the higher force constant cantilevers. Cantilevers having force constants of 0.9 and 3 N m(-1) demonstrated minimal vertical sample compression with increasing applied force. The effects of AFM image pixel density and scan speed on the measured height (diameter) of SWNTs were also assessed.     

The influence of gravity on the distribution of the deposit formed onto a substrate by sessile, hanging, and sandwiched hanging drop evaporation

The focus of the present article is the study of the influence of gravity on the particle deposition profiles on a solid substrate during the evaporation of sessile, hanging and sandwiched hanging drops of colloidal particle suspensions. For concentrations of nanoparticles in the colloidal solutions in the range 0.0001-1 wt.%, highly diluted suspensions will preferentially form rings while concentrated suspensions will preferentially form spots in both sessile and hanging drop evaporation. For intermediary concentrations, the particle deposition profiles will depend on the nanoparticle aggregation dynamics in the suspension during the evaporation process, gravity and on the detailed evaporation geometry. The evaporation of a drop of toluene/carbon nanoparticle suspension hanging from a pendant water drop will leave on the substrate a circular spot with no visible external ring. By contrast, a clear external ring is formed on the substrate by the sessile evaporation of a similar drop of suspension sandwiched between a water drop and the substrate. From the application viewpoint, these processes can be used to create preferential electrical conductive carbon networks and contacts for arrays of self-assembled nanostructures fabricated on solid substrates as well as on flexible polymeric substrates.