Evidence for Enhanced Tracer Diffusion in Densely Packed Interfacial Assemblies of Hairy Nanoparticles.

Nearly monodisperse nanoparticle (NP) spheres attached to a nonvolatile ionic liquid surface were tracked by in situ scanning electron microscopy to obtain the tracer diffusion coefficient Dtr as a function of the areal fraction ϕ. The in situ technique resolved both tracer (gold) and background (silica) particles for ∼1-2 min, highlighting their mechanisms of diffusion, which were strongly dependent on ϕ. Structure and dynamics at low and moderate ϕ paralleled those reported for larger colloidal spheres, showing an increase in order and a decrease in Dtr by over 4 orders of magnitude. However, ligand interactions were more important near jamming, leading to different caging and jamming dynamics for smaller NPs. The normalized Dtr at ultrahigh ϕ depended on particle diameter and ligand molecular weight. Increasing the PEG molecular weight by a factor of 4 increased Dtr by 2 orders of magnitude at ultrahigh ϕ, indicating stronger ligand lubrication for smaller particles.

Wetting, meniscus structure, and capillary interactions of microspheres bound to a cylindrical liquid interface.

Wetting, meniscus structure, and capillary interactions for polystyrene microspheres deposited on constant curvature cylindrical liquid interfaces, constructed from nonvolatile ionic or oligomeric liquids, were studied by optical interferometry and optical microscopy. The liquid interface curvature resulted from the preferential wetting of finite width lines patterned onto planar silicon substrates. Key variables included sphere diameter, nominal (or average) contact angle, and deviatoric interfacial curvature. Menisci adopted the quadrupolar symmetry anticipated by theory, with interfacial deformation closely following predicted dependences on sphere diameter and nominal contact angle. Unexpectedly, the contact angle was not constant locally around the contact line, the nominal contact angle varied among seemingly identical spheres, and the maximum interface deviation did not follow the predicted dependence on deviatoric interfacial curvature. Instead, this deviation was up to an order-of-magnitude larger than predicted. Trajectories of neighboring microspheres visually manifested quadrupole-quadrupole interactions, eventually producing square sphere packings that foreshadow interfacial assembly as a potential route to hierarchical 2D particle structures.

Lysozyme Solubility and Conformation in Neat Ionic Liquids and Their Mixtures with Water.

The room temperature solubility of a number of model proteins is assessed for a diverse set of neat ionic liquids (ILs). For two soluble protein-IL pairs, lysozyme in [C2MIM][EtSO4] (1-ethyl-3-methylimidazolium ethylsulfate) and in [C2,4,4,4P][Et2PO4] (tributyl(ethyl)phosphonium diethylphosphate), protein solubility and structure at various temperatures are probed by dynamic light scattering (assessing dissolved molecular size), turbidimetry (reflecting degree of solubility), and Fourier transform infrared spectroscopy (uncovering helical secondary structure). As compared to aqueous environments, [C2,4,4,4P][Et2PO4] thermally stabilizes protein size and secondary structure while [C2MIM][EtSO4] does the opposite. Lysozyme denatured in [C2MIM][EtSO4] does not aggregate, presumably due to an absence of hydrophobic interactions, and the denaturation appears thermally reversible. Both ILs at room temperature are miscible with water in all proportions, but to create the corresponding ternary mixtures with protein, the order of mixing is important. Mixed to avoid additions of water to IL-dissolved protein, stable solutions are obtained with [C2MIM][EtSO4] at all solvent compositions. When water is added to IL-rich solutions, liquid-liquid demixing is noted.

Interfacial rheology of polymer/carbon nanotube films co-assembled at the oil/water interface.

At appropriate conditions, water-dispersed acid-functionalized single-walled carbon nanotubes (SWCNTs) co-assemble at the oil/water interface with toluene-dissolved amine-terminated polystyrene (PS-NH2) to form composite thin films displaying pronounced interfacial viscoelasticity. To probe this viscoelasticity, the films were examined under dilatational deformations of pendant drop tensiometry/rheometry, with storage and loss moduli recorded against frequency ω (0.003 < ω < 3 Hz) and time-dependent relaxation modulus recorded against time t (0.2 < t < 2000 s). Without the SWCNTs, PS-NH2-decorated interfaces have little dilatational stiffness, i.e., low storage modulus, but their stiffness grows as SWCNTs are added, reaching 50-100 mN m-1 at large ω. Two characteristic relaxation processes are identified in the composite films: a fast process (ω ∼ 0.1-0.2 Hz) attributable to local structural relaxation of confined PS-NH2 and a slow process (t ∼ 300-2000 s) attributable to component adsorption/desorption (or attachment/detachment). Among the variables that affect positions and strengths of these relaxations are SWCNT and PS-NH2 bulk concentrations as well as water phase pH. In frequency or timescale ranges intermediate between the two relaxations, the co-assembled films display "soft-glass" behavior, with the storage and loss moduli characterized by nearly equal power-law exponents. The relaxation modulus, better able to probe terminal behavior, eventually decays to zero, revealing that the films are fundamentally fluid-like due to the slow relaxation, and in support of this conclusion, large strain compression-induced film wrinkles disappear at large t.

Nonfreezing Water Structuration in Heteroprotein Coacervates.

Surface-bound water in protein solutions has been identified with a reduction in its freezing point. We studied the presence of such nonfreezing water (NFW) in various protein-polyelectrolyte, micelle-polyelectrolyte, and protein-protein (heteroprotein) coacervates, along with appropriate concentrated solutions of macromolecules alone, finding up to 15% w/w NFW for the heteroprotein coacervate of lactoferrin (LF) and ß-lactoglobulin (BLG). The level of NFW is always higher in coacervates than in the control (single macromolecule) systems, particularly for protein-containing coacervates: a coacervate of bovine serum albumin (BSA) and poly(dimethyldiallylammonium chloride) (PDADMAC) showed a ratio of NFW/protein twice that of BSA alone (0.6 vs 0.3), with a similarly high ratio for LF-BLG coacervate. These results are attributed to the maximization of water-protein contacts, structural features that reflect the mode of sample assembly, as they are not seen in a noncoacervated LF-BLG solution with identical concentrations of all species.

Assembly of acid-functionalized single-walled carbon nanotubes at oil/water interfaces.

The efficient segregation of water-soluble, acid-functionalized, single-walled carbon nanotubes (SWCNTs) at the oil/water interface was induced by dissolving low-molecular-weight amine-terminated polystyrene (PS-NH2) in the oil phase. Salt-bridge interactions between carboxylic acid groups of SWCNTs and amine groups of PS drove the assembly of SWCNTs at the interface, monitored by pendant drop tensiometry and laser scanning confocal microscopy. The impact of PS end-group functionality, PS and SWCNT concentrations, and the degree of SWCNT acid modification on the interfacial activity was assessed, and a sharp drop in interfacial tension was observed above a critical SWCNT concentration. Interfacial tensions were low enough to support stable oil/water emulsions. Further experiments, including potentiometric titrations and the replacement of SWCNTs by other carboxyl-containing species, demonstrated that the interfacial tension drop reflects the loss of SWCNT charge as the pH falls near/below the intrinsic carboxyl dissociation constant; species lacking multivalent carboxylic acid groups are inactive. The trapped SWCNTs appear to be neither ordered nor oriented.

Protein-selective coacervation with hyaluronic acid.

Selective coacervation with hyaluronic acid (HA), a biocompatible and injectable anionic polysaccharide, was used to isolate a target protein, bovine serum albumin (BSA), with 90% purity from a 1:1 mixture with a second protein of similar pI, ß-lactoglobulin (BLG). This separation was attributed to the higher HA-affinity of BSA, arising from its more concentrated positive domain. The values of pH corresponding respectively to the onset of complex formation, coacervation, precipitation, and redissolution (pH(c), pHϕ, pH(p), and pH(d)) were determined as a function of ionic strength I. These pH values were related to critical values of protein charge, Z, and their dependence on I provided some insights into the mechanisms of these transitions. The higher polyanion binding affinity of BSA, deduced from its higher values of pH(c), was confirmed by isothermal titration calorimetry (ITC). Confocal laser microscopy clearly showed time-dependent coalescence of vesicular droplets into a continuous film. Comparisons with prior results for the polycation poly(diallyldimethylammonium chloride) (PDADMAC) show reversal of protein selectivity due to reversal of the polyelectrolyte charge. Stronger binding of both proteins to PDADMAC established by ITC may be related to the higher chain flexibility and effective linear charge density of this polycation.

Ionic liquids as floatation media for cryo-ultramicrotomy of soft polymeric materials.

Ionic liquids (ILs) and their mixtures with low molecular solvents present ideal properties for use as flotation liquids in cryo-ultramicrotomy. With control of T g and η by co-solvent addition, flat, ultra-thin sections are reliably floated onto transmission electron microscopy grids even at temperatures as low as -100°C. Even more, the liquids and their mixtures are stable in the microtome trough for several hours because of low vapor pressure and low solidification temperature. Compared to established flotation media for soft polymer systems, the time and skill needed for cryo-ultramicrotomy are significantly reduced. Although just a handful of ILs are discussed and a good general choice identified, if different liquid characteristics are needed for a particular sample, thousands of additional ILs will perform similarly, giving this new approach enormous flexibility.

Dynamic Reconfiguration of Compressed 2D Nanoparticle Monolayers.

A Gibbs monolayer of jammed, or nearly jammed, spherical nanoparticles was imaged at a liquid surface in real time by in-situ scanning electron microscopy performed at the single-particle level. At nanoparticle areal fractions above that for the onset of two-dimensional crystallization, structural reorganizations of the mobile polymer-coated particles were visualized after a stepwise areal compression. When the compression was small, slow shearing near dislocations and reconfigured nanoparticle bonding were observed at crystal grain boundaries. At larger scales, domains grew as they rotated into registry by correlated but highly intermittent motions. Simultaneously, the areal density in the middle of the monolayer increased. When the compression was large, the jammed monolayers exhibited out-of-plane deformations such as wrinkles and bumps. Due to their large interfacial binding energy, few (if any) of the two-dimensionally mobile nanoparticles returned to the liquid subphase. Compressed long enough (several hours or more), monolayers transformed into solid nanoparticle films, as evidenced by their cracking and localized rupturing upon subsequent areal expansion. These observations provide mechanistic insights into the dynamics of a simple model system that undergoes jamming/unjamming in response to mechanical stress.

Dissolution and dissolved state of cytochrome C in a neat, hydrophilic ionic liquid.

The dissolution and dissolved molecular state of cytochrome c were investigated in the room temperature ionic liquid ethylmethylimidazolium ethylsulfate, [EMIM][EtSO4], by viscometry, optical and vibrational spectroscopies, and peroxidase activity. In dilute mixtures, viscometry demonstrated true molecular dissolution of cytochrome c in the ionic liquid and uncovered a molecular size larger than that in aqueous buffer, suggesting altered solvation or slight denaturation. The protein's heme unit absorbs light outside the spectral range masked by [EMIM], enabling conformational assessments by UV-visible and circular dichroism spectroscopies. Adding trends from fluorescence and Fourier transform infrared spectroscopy, unchanged secondary but perturbed tertiary structures were determined, consistent with the appreciable peroxidase activity measured. Different than in aqueous buffers, denaturation is not accompanied by aggregation. Results are relevant to the proposed application of ionic liquids as media for room temperature preservation of biomacromolecules.

Bidisperse Nanospheres Jammed on a Liquid Surface.

Jammed packings of bidisperse nanospheres were assembled on a nonvolatile liquid surface and visualized to the single-particle scale by using an in situ scanning electron microscopy method. The PEGylated silica nanospheres, mixed at different number fractions and size ratios, had large enough in-plane mobilities prior to jamming to form uniform monolayers reproducibly. From the collected nanometer-resolution images, local order and degree of mixing were assessed by standard metrics. For equimolar mixtures, a large-to-small size ratio of about 1.5 minimized correlated metrics for local orientational and positional order, as previously predicted in simulations of bidisperse disk jamming. Despite monolayer uniformity, structural and depletion interactions caused spheres of a similar size to cluster, a feature evident at size ratios above 2. Uniform nanoparticle monolayers of high packing disorder are sought in many liquid interface technologies, and these experiments outlined key design principles, buttressing extensive theory/simulation literature on the topic.

Diffusion of circular DNA in two-dimensional cavity arrays.

Through a two-dimensional cavity array with connecting pores of submolecular size, diffusion of relaxed circular and linear DNA molecules is visualized by fluorescence microscopy. Across the entropic barriers transport regime, associated with spatially heterogeneous confinement of flexible polymers, circular DNA diffuses slower than linear DNA of the same length, a trend indicating that linear DNA preferably moves through connecting pores by the threading of an end rather than the looping of a midsection.

Assessing Pair Interaction Potentials of Nanoparticles on Liquid Interfaces.

The pair interaction potentials of polymer-grafted silica nanoparticles (NPs) at liquid surfaces were determined by scanning electron microscopy, exploiting the nonvolatility of ionic liquids to stabilize the specimens against microscope vacuum. Even at near contact, individual, two-dimensionally well-dispersed NPs were resolved. The potential of mean force, reduced to the pair interaction potential for dilute NPs, was extracted with good accuracy from the radial distribution function, as both NP diameter and grafted polymer chain length were varied. While NP polydispersity somewhat broadened the core repulsion, the pair potential well-approximated a hard sphere interaction, making these systems suitable for model studies of interfacially bound NPs. For short (5 kDa) poly(ethylene glycol) ligands, a weak (< kB T) long-range attraction was discerned, and for ligands of identical length, pair potentials overlapped for NPs of different diameter; the attraction is suggested to arise from ligand-induced menisci. To understand better the interactions underlying the pair potential, NP surface-binding energies were measured by interfacial tensiometry, and NP contact angles were assessed by atomic force microscopy and transmission electron microscopy.

Visualizing the Dynamics of Nanoparticles in Liquids by Scanning Electron Microscopy.

Taking advantage of ionic liquid nonvolatility, the Brownian motions of nanospheres and nanorods in free-standing liquid films were visualized in situ by scanning electron microscopy. Despite the imaging environment's high vacuum, a liquid cell was not needed. For suspensions that are dilute and films that are thick compared to the particle diameter, the translational and rotational diffusion coefficients determined by single-particle tracking agree with theoretical predictions. In thinner films, a striking dynamical pairing of nanospheres was observed, manifesting a balance of capillary and hydrodynamic interactions, the latter strongly accentuated by the two-dimensional film geometry. Nanospheres at high concentration displayed subdiffusive caged motion. Concentrated nanorods in the thinner films transiently assembled into finite stacks but did not achieve high tetratic order. The illustrated imaging protocol will broadly apply to the study of soft matter structure and dynamics with great potential impact.

Ionic liquids as plasticizers for polyelectrolyte complexes.

Uptake of salts by insoluble polyelectrolyte complexes (PECs) leads to plasticization, and here it is shown that ionic liquids (ILs) are more effective plasticizers than simple organic salts such as NaCl. The PEC uptake of IL cation was monitored by solution (1)H NMR, and the mechanical impacts of plasticization were tracked by dynamic mechanical analysis (DMA). PECs prepared with polystyrene sulfonate (PSS) and poly(diallyldimethylammonium chloride) (PDDA) under charge stoichiometric conditions were immersed in aqueous solutions of 1-butyl-3-methylimidazolium chloride [BMIM][Cl] to cause IL uptake, which could be controlled by the solution's IL concentration: higher concentration leads to higher uptake which leads to greater plasticization. The effectiveness of plasticization was assessed through the position and height of a DMA tan(δ) peak ascribed to a glassy-to-rubbery PEC transition. Consistent with greater PEC uptake, isothermal titration calorimetry demonstrated that solution binding by PSS of [BMIM](+) was much stronger than binding of Na(+).

Thermoreversible gelation of an ionic liquid by crystallization of a dissolved polymer.

Poly(ethylene glycol) (PEG) dissolves in the room-temperature ionic liquid 1-ethyl-3-methylimidazolium ethylsulfate [EMIM][EtSO(4)] above approximately 60 degrees C, the neat polymer's melting temperature, and if polymer concentration and molecular weight are high enough, the solution transforms into a semitransparent gel when cooled. The modulus, reaching 100 KPa or higher, is strongly affected by PEG concentration, and self-supporting materials are made even from solutions somewhat below coil overlap. Via differential scanning calorimetry (DSC), rheology, and optical microscopy, thermoreversible solidification is traced to kinetically frustrated polymer crystallization, an established mechanism for many pairings of crystallizable polymer with aqueous or organic solvent. Optical microscopy reveals nucleation and growth of PEG crystals with a largest dimension of tens to hundreds of micrometers. Crystalline chain packing in gels is identical to that of neat PEG, and degrees of crystallization are similar. Simple preparation, nontoxicity, and vanishing volatility suggest unique new gel applications.

Brownian motion of DNA confined within a two-dimensional array.

Linear DNA molecules are visualized while undergoing Brownian motion inside media patterned with molecular-sized spatial constraints. The media, prepared by colloidal templating, trap the macromolecules within a two-dimensional array of spherical cavities interconnected by circular holes. Across a broad DNA size range, diffusion does not proceed by the familiar mechanisms of reptation or sieving. Rather, because of their inherent flexibility, DNA molecules strongly localize in cavities and only sporadically "jump" through holes. Jumping closely follows Poisson statistics. By reducing DNA's configurational freedom, the holes act as molecular weight-dependent entropic barriers. Sterically constrained macromolecular diffusion underlies many separation methods and assumes an important role in intracellular and extracellular transport.