KyuHan Kim

Interfacial microrheology of DPPC monolayers at the air–water interface

We present systematic measurements of the surface rheology of monolayers of liquid-condensed (LC) dipalmitoylphosphatidylcholine (DPPC) at the air–water interface. Using microfabricated, ferromagnetic ‘microbuttons’ as new microrheological probes, we measure the linear viscoelastic moduli of LC DPPC monolayers as both surface pressure and frequency are varied. Visualization of this interface reveals that the interlocked liquid crystalline domains that comprise an LC-DPPC monolayer give rise to a viscoelastic solid response. Two distinct behaviors arise as surface pressure is increased: for low surface pressures (8 mN m−1 ≤ Π ≤ 12–14 mN m−1), the monolayer behaves like a two-dimensional emulsion, with a surface elastic modulus G ′s that is relatively constant, as would be expected from a line tension-mediated elasticity. The surface viscosity increases exponentially with Π, as would be expected for a condensed liquid monolayer. Above 12–14 mN m−1, however, both moduli increase exponentially with Π, albeit with a weaker slope—a response that would not be expected from line-tension-mediated elasticity. This transition would be consistent with a second-order phase transition between the LC and solid-condensed phase, as has been observed in other phospholipid monolayers. Finally, we employ a controlled-stress (creep) mode to find a stress-dependent viscosity bifurcation, and thus the yield stress of this monolayer.

Effect of cholesterol nanodomains on monolayer morphology and dynamics

Significance Replacement lung surfactants have dramatically reduced premature infant mortality owing to respiratory distress syndrome. However, clinical lung surfactant varies widely in composition, and even the existence of cholesterol in native lung surfactants remains controversial. Improving replacement surfactants will require an understanding of each molecular component’s role in a film’s static and dynamic properties. Here, we show that small cholesterol fractions reduce the viscosity of model lung surfactant interfaces by orders of magnitude while leaving compressibility and collapse unchanged, offering control over surfactant spreadability. Lipid–cholesterol nanodomain complexes are observed, which act as line-active sources of free area to reduce surface viscosity. At low mole fractions, cholesterol segregates into 10- to 100-nm-diameter nanodomains dispersed throughout primarily dipalmitoylphosphatidylcholine (DPPC) domains in mixed DPPC:cholesterol monolayers. The nanodomains consist of 6:1 DPPC:cholesterol “complexes” that decorate and lengthen DPPC domain boundaries, consistent with a reduced line tension, λ. The surface viscosity of the monolayer, ηs, decreases exponentially with the area fraction of the nanodomains at fixed surface pressure over the 0.1- to 10-Hz range of frequencies common to respiration. At fixed cholesterol fraction, the surface viscosity increases exponentially with surface pressure in similar ways for all cholesterol fractions. This increase can be explained with a free-area model that relates ηs to the pure DPPC monolayer compressibility and collapse pressure. The elastic modulus, G′, initially decreases with cholesterol fraction, consistent with the decrease in λ expected from the line-active nanodomains, in analogy to 3D emulsions. However, increasing cholesterol further causes a sharp increase in G′ between 4 and 5 mol% cholesterol owing to an evolution in the domain morphology, so that the monolayer is elastic rather than viscous over 0.1–10 Hz. Understanding the effects of small mole fractions of cholesterol should help resolve the controversial role cholesterol plays in human lung surfactants and may give clues as to how cholesterol influences raft formation in cell membranes.

Processable high internal phase Pickering emulsions using depletion attraction

High internal phase emulsions have been widely used as templates for various porous materials, but special strategies are required to form, in particular, particle-covered ones that have been more difficult to obtain. Here, we report a versatile strategy to produce a stable high internal phase Pickering emulsion by exploiting a depletion interaction between an emulsion droplet and a particle using water-soluble polymers as a depletant. This attractive interaction facilitating the adsorption of particles onto the droplet interface and simultaneously suppressing desorption once adsorbed. This technique can be universally applied to nearly any kind of particle to stabilize an interface with the help of various non- or weakly adsorbing polymers as a depletant, which can be solidified to provide porous materials for many applications.

Influence of molecular coherence on surface viscosity.

Adding small fractions of cholesterol decreases the interfacial viscosity of dipalmitoylphosphatidylcholine (DPPC) monolayers by an order of magnitude per wt %. Grazing incidence X-ray diffraction shows that cholesterol at these small fractions does not mix ideally with DPPC but rather induces nanophase separated structures of an ordered, primarily DPPC phase bordered by a line-active, disordered, mixed DPPC-cholesterol phase. We propose that the free area in the classic Cohen and Turnbull model of viscosity is inversely proportional to the number of molecules in the coherence area, or product of the two coherence lengths. Cholesterol significantly reduces the coherence area of the crystals as well as the interfacial viscosity. Using this free area collapses the surface viscosity data for all surface pressures and cholesterol fractions to a universal logarithmic relation. The extent of molecular coherence appears to be a fundamental factor in determining surface viscosity in ordered monolayers.

Linear and nonlinear microrheometry of small samples and interfaces using microfabricated probes

We describe a microrheological strategy that enables sensitive surface shear rheology measurements of surfactant-laden interfaces, with the capacity to simultaneously visualize deforming interfaces. This technique utilizes a ferromagnetic microbutton probe pinned to a fluid-fluid interface, and actively torqued or forced with externally controlled electromagnets. Various modes of operation are possible: Small-amplitude oscillatory rotations, which provide frequency-dependent viscoelastic shear moduli; controlled torque (analogous to fixing shear stress); controlled rotation rate (analogous to fixing strain rate); and imposed force (analogous to active, translational microrheology). The circular shape of the probe ensures pure shear strains (when driven to rotate). We describe the experimental apparatus, its measurement limits and sources of error. We then highlight its versatility and capabilities with measurements on a variety of qualitatively distinct systems, including purely viscous monolayers, block-...

Surface charge regulation of carboxyl terminated polystyrene latex particles and their interactions at the oil/water interface.

We study electrostatic interactions of polystyrene particles at an oil/water interface controlled by a chemical reaction of carboxylate surface functional groups. By replacing the carboxyl functional groups with hydrocarbon chains using the well-known EDC (1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide) coupling reaction, the surface charge density decreases while the hydrophobicity of the colloid surface increases. Direct visualization of the particle-laden interface reveals that, depending on the extent of hydrocarbon coupling, the strength of the electrostatic repulsion can be regulated: the repulsive interaction increases with the reaction, removing aggregates, but rapidly decreases if the reaction proceeds too much, forming a large aggregation. This simple reaction, thus, dramatically changes the structures of the colloidal monolayers at the oil/water interface. We conclude that such structural change is the result of change of the repulsive interactions from the oil phase, although interactions in the water phase are also changed slightly.

Fluorescence Recovery after Merging a Surfactant-Covered Droplet: A Novel Technique to Measure the Diffusion of Phospholipid Monolayers at Fluid/Fluid Interfaces

We present a novel technique to measure diffusion coefficients of insoluble surfactant monolayers. We merge a surfactant-coated droplet with a fluorescently labeled planar monolayer. During the merging process, a monolayer on a droplet displaces the existing planar monolayer, leaving a dark area when viewed under a fluorescence microscope. We measure fractional intensities as the dyes recover, which allows diffusion coefficients to be computed. We validate this technique with the two most common phospholipid monolayers (DPPC and DOPC) and study the diffusion of their mixtures. The proposed technique has several advantages over the FRAP technique and is potentially capable of measuring the diffusion of any soluble/insoluble surfactant monolayers.

Static and Dynamic Permeability Assay for Hydrophilic Small Molecules Using a Planar Droplet Interface Bilayer

Because numerous drugs are administered through an oral route and primarily absorbed at the intestine, the prediction of drug permeability across an intestinal epithelial cell membrane has been a crucial issue in drug discovery. Thus, various in vitro permeability assays have been developed such as the Caco-2 assay, the parallel artificial membrane permeability assay (PAMPA), the phospholipid vesicle-based permeation assays (PVPA) and Permeapad. However, because of the time-consuming and quite expensive process for culturing cells in the Caco-2 assay and the unknown microscopic membrane structures of the other assays, a simpler yet more accurate and versatile technique is still required. Accordingly, we developed a new platform to measure the permeability of small molecules across a planar freestanding lipid bilayer with a well-defined area and structure. The lipid bilayer was constructed within a conventional UV spectrometer cell, and the transport of drug molecules across the bilayer was recorded by UV absorbance over time. We then computed the permeability from the time-dependent diffusion equation. We tested this assay for five exemplary hydrophilic drugs and compared their values with previously reported ones. We found that our assay has a much higher permeability compared to the other techniques, and this higher permeability is related to the thickness of the lipid bilayer. Also we were able to measure the dynamic permeability upon the addition of a membrane-disrupting surfactant demonstrating that our assay has the capability to detect real-time changes in permeability across the lipid bilayer.

Fluorescence Recovery after Merging a Droplet to Measure the Two- dimensional Diffusion of a Phospholipid Monolayer

We introduce a new method to measure the lateral diffusivity of a surfactant monolayer at the fluid-fluid interface, called fluorescence recovery after merging (FRAM). FRAM adopts the same principles as the fluorescence recovery after photobleaching (FRAP) technique, especially for measuring fluorescence recovery after bleaching a specific area, but FRAM uses a drop coalescence instead of photobleaching dye molecules to induce a chemical potential gradient of dye molecules. Our technique has several advantages over FRAP: it only requires a fluorescence microscope rather than a confocal microscope equipped with high power lasers; it is essentially free from the selection of fluorescence dyes; and it has far more freedom to define the measured diffusion area. Furthermore, FRAM potentially provides a route for studying the mixing or inter-diffusion of two different surfactants, when the monolayers at a surface of droplet and at a flat air/water interface are prepared with different species, independently.