Frequency-Dependent Microelectrophoresis Study of Colloids with Tunable Surface Charge.

Nonaqueous poly(methyl methacrylate) (PMMA) colloidal suspensions in a solvent that is simultaneously matched in both density and refractive index have been important for real-space studies of colloidal self-assembly, but their complex electrostatic character remains largely unexplored. Electrophoresis is a powerful tool for determining the surface potential and charge of the colloidal suspension; however, because of refractive index matching, standard electrophoresis measurements are not feasible. We carry out microscope-based microelectrophoresis measurements on PMMA colloids in cyclohexyl bromide and cis-trans decalin to measure particle charge as a function of salt concentration in both DC and frequency-variable AC fields. The colloid charge depends on salt concentration and reverses sign near 0.35 µM, providing evidence that solution ions are actively modifying the colloid surface. The frequency dependence of the electrophoretic mobility yields the characteristic time scale for electric double-layer polarization and provides intriguing evidence for Manning condensation and polyion formation.

The effects of biological crowders on fibrillization, structure, diffusion, and conformational dynamics of α-synuclein.

α-synuclein is an intrinsically disordered protein (IDP) whose aggregation in presynaptic neuronal cells is a pathological hallmark of Lewy body formation and Parkinson's disease. This aggregation process is likely affected by the crowded macromolecular cellular environment. In this study, α-synuclein was studied in the presence of both a synthetic crowder, Ficoll70, and a biological crowder composed of lysed cells that better mimics the biocomplexity of the cellular environment. 15 N-1 H HSQC NMR results show similar α-synuclein chemical shifts in non-crowded and all crowded conditions implying that it remains similarly unstructured in all conditions. Nevertheless, both HSQC NMR and fluorescence measurements indicate that, only in the cell lysate, α-synuclein forms aggregates over a timescale of 48 h. 15 N-edited diffusion measurements indicate that all crowders slow down the α-synuclein's diffusivity. Interestingly, at high concentrations, α-synuclein diffuses faster in cell lysate than in Ficoll70, possibly due to additional soft (e.g., electrostatic or hydrophobic) interactions. 15 N-edited relaxation measurements show that some residues are more mobile in cell lysate than in Ficoll70; the rates that are most different are predominantly in hydrophobic residues. We thus examined cell lysates with reduced hydrophobicity and found slower dynamics (higher relaxation rates) in several α-synuclein residues. Taken together, these experiments suggest that while cell lysate does not substantially affect α-synuclein structure (HSQC spectra), it does affect chain dynamics and translational diffusion, and strongly affects aggregation over a timescale of days, in a manner that is different from either no crowder or an artificial crowder: soft hydrophobic interactions are implicated.

Concentration Dependence of Elastic and Viscoelastic Properties of Aqueous Solutions of Ficoll and Bovine Serum Albumin by Brillouin Light Scattering Spectroscopy.

The cellular environment is crowded with macromolecules of different shapes and sizes. The effect of this macromolecular crowding has been studied in a variety of synthetic crowding environments: two popular examples are the compact colloid-like Ficoll macromolecule and the globular protein bovine serum albumin (BSA). Recent studies have indicated that a significant component of bound or surface-associated water in these crowders reduces the available free volume. In this work, Brillouin light scattering experiments were performed on aqueous solutions of Ficoll 70 and Ficoll 400 with concentrations ranging from 1 to 35 wt % and BSA with concentrations of 1 to 27 wt %. From the dependence of spectral peak parameters on polymer concentration, we determined fundamental solution properties: hypersound velocity, adiabatic bulk modulus and compressibility, apparent viscosity, and hypersound attenuation. The existing theory that ignores intermolecular interactions can capture only the observed linear trends in the frequency shift up to a threshold concentration, beyond which a quadratic term accounting for intermolecular interactions is necessary. This likely indicates a transition from the dilute to semidilute regime. In the Ficoll solutions (but not BSA), we see evidence for a central mode, which is indicative of relaxation in the hydration shell of Ficoll.

Diffusion NMR and Rheology of a Model Polymer in Bacterial Cell Lysate Crowders.

The intracellular milieu is crowded and heterogeneous, and this can have profound consequences for biomolecule motions and biochemical kinetics. Macromolecular crowding has been traditionally studied in artificial crowders like Ficoll and dextran or globular proteins such as bovine serum albumin. It is, however, not clear if the effects of artificial crowders on such phenomena are the same as the crowding that is experienced in a heterogeneous biological environment. Bacterial cells, for example, are composed of heterogeneous biomolecules with different sizes, shapes, and charges. Using crowders composed of one of three different pretreatments of bacterial cell lysate (unmanipulated, ultracentrifuged, and anion exchanged), we examine the effects of crowding on the diffusivity of a model polymer. We measure the translational diffusivity, via diffusion NMR, of the test polymer polyethylene glycol (PEG) in these bacterial cell lysates. We show that the small (Rg ∼ 5 nm) test polymer shows a modest decrease in self-diffusivity with increasing crowder concentration for all lysate treatments. The corresponding self-diffusivity decrease in the artificial Ficoll crowder is much more pronounced. Moreover, a comparison of the rheological response of biological and artificial crowders shows that while the artificial crowder Ficoll exhibits a Newtonian response even at high concentrations, the bacterial cell lysate is markedly non-Newtonian; it behaves like a shear-thinning fluid with a yield stress. While at any concentration the rheological properties are sensitive to both lysate pretreatment and batch-to-batch variations, the PEG diffusivity is nearly unaffected by the type of lysate pretreatment.

Frequency-dependent viscosity of salmon ovarian fluid has biophysical implications for sperm-egg interactions.

Gamete-level sexual selection of externally fertilising species is usually achieved by modifying sperm behaviour with mechanisms that alter the chemical environment in which gametes perform. In fish, this can be accomplished through the ovarian fluid, a substance released with the eggs at spawning. While the biochemical effects of ovarian fluid in relation to sperm energetics have been investigated, the influence of the physical environment in which sperm compete remains poorly explored. Our objective was therefore to gain insights on the physical structure of this fluid and potential impacts on reproduction. Using soft-matter physics approaches of steady-state and oscillatory viscosity measurements, we subjected wild Atlantic salmon ovarian fluids to variable shear stresses and frequencies resembling those exerted by sperm swimming through the fluid near eggs. We show that this fluid, which in its relaxed state is a gel-like substance, displays a non-Newtonian viscoelastic and shear-thinning profile, where the viscosity decreases with increasing shear rates. We concurrently find that this fluid obeys the Cox-Merz rule below 7.6 Hz and infringes it above this level, thus indicating a shear-thickening phase where viscosity increases provided it is probed gently enough. This suggests the presence of a unique frequency-dependent structural network with relevant implications for sperm energetics and fertilisation dynamics. This article has an associated ECR Spotlight interview with Marco Graziano.

2H NMR Study of Polymer Segmental Dynamics at Varying Cross-Linking in Poly(N-isopropylacrylamide) Microgels.

A microscopic understanding of the internal structure and dynamics of poly(N-isopropylacrylamide) (PNIPAM) chains, in microgel colloids, is developed using deuterium NMR (2H NMR) to study deuterated PNIPAM suspensions as functions of temperature and pressure for four cross-linker molar fractions (f). The PNIPAM polymers were labeled with deuterons at the backbone (d3-PNIPAM) or on side chains (d7-PNIPAM). 2H NMR spectra of the d3-PNIPAM suspensions for all cross-linker molar fractions indicated freely moving chains at low temperature and a nearly immobilized fraction above ∼35 °C. Polymer segments in the collapsed phase of the d3-PNIPAM suspension were more mobile than those in the dry powder. This is direct microscopic evidence that the polymer remains significantly hydrated in the collapsed phase, consistent with strong, indirect evidence from recent light scattering and rheology measurements from our laboratory. However, the observation of a small fraction of immobilized segments in the swollen phase for higher cross-linker molar fraction suggests that, particularly for high levels of cross-linking, some polymer is nonhydrated even in the swollen phase. Finally, variable-pressure NMR (up to 90 MPa) showed a slight increase in transition temperature with pressure for lower cross-linker molar fractions and a larger increase in transition temperature with pressure for higher cross-linker molar fractions. This is consistent with a previously reported dependence of collapse transition enthalpy on cross-linker molar fraction.

Structural parameters of soft PNIPAM microgel particles as a function of crosslink density.

HYPOTHESIS: The temperature dependences of hydrodynamic radii in thermo-sensitive microgel suspensions, known as collapse curves, are commonly fitted to the benchmark Flory-Rehner theory but parameters obtained often yield little physical insights. Our study of poly(N-isopropylacrylamide) (PNIPAM) microgel suspensions in water is driven by the hypothesis that fitting to Flory-Rehner theory can yield meaningful parameters that separate into ones that are insensitive to crosslink density or deuteration and ones that are not. EXPERIMENTS: Dynamic light scattering (DLS) and rheology experiments were done on 8 microgel variants, protonated and deuterated PNIPAM for four crosslink densities, synthesized under otherwise identical conditions. FINDINGS: Remarkably, polymer volume fractions in the microgel particle at collapse, ϕcollapse, obtained via rheology, are independent of crosslink density. Along with collapse curves from DLS, this determines the temperature dependence of microgel water and polymer volume fractions. Fitting collapse curves to Flory-Rehner theory yields reference polymer volume fractions, ϕ0, associated with microgel particle elasticity. ϕ0 is much lower than ϕcollapse, and increases with crosslink density. For all microgel sample variants, a crossover temperature, where the elastic contribution to osmotic pressure changes sign, is found to approximate the final temperature after microgel synthesis and also to the free polymer θ temperature.

Correction: Tunable hydrodynamics: a field-frequency phase diagram of a non-equilibrium order-to-disorder transition.

Correction for 'Tunable hydrodynamics: a field-frequency phase diagram of a non-equilibrium order-to-disorder transition' by Somayeh Khajehpour Tadavani et al., Soft Matter, 2017, 13, 7412-7424.

Anomalous dynamics in tracer-particle motions in an electrohydrodynamically driven oil-in-oil system.

We characterize the superdiffusive dynamics of tracer particles in an electrohydrodynamically driven emulsion of oil droplets in an immiscible oil medium, where the amplitude and frequency of an external electric field are the control parameters. In the weakly driven electrohydrodynamic regime, the droplets are trapped dielectrophoretically on a patterned electrode, and the driving is therefore spatially varying. We find excellent agreement with a 〈x^{2}〉∼t^{1.5} power law and find that this superdiffusive dynamics arises from an underlying displacement distribution that is distinctly non-Gaussian and exponential for small displacements and short times. While these results are comparable with a random-velocity field model, the tracer particle speeds are in fact spatially varying in two dimensions, arising from a spatially varying electrohydrodynamic driving force. This suggests that the important ingredient for the superdiffusive t^{1.5} behavior observed is a velocity field that is isotropic in the plane and spatially correlated. Finally, we can extract, from the superdiffusive dynamics, a experimental length scale that corresponds to the lateral range of the hydrodynamic flows. This experimental length scale is non zero only above a threshold ion mobility length.

Realization of a stable, monodisperse water-in-oil droplet system with micro-scale and nano-scale confinement for tandem microscopy and diffusion NMR studies.

In this work we generate stable and monodisperse water-in-oil emulsions using a co-flowing geometry that produced droplet sizes between 13 µm and 250 µm. The drops survived transfer to NMR tubes and were stable for at least 26 hours, enabling the performance of pulsed-field-gradient NMR experiments in addition to microscopy. The drops sizes achieved as a function of flow rate agree well with a simple model for droplet generation: this yields a precise measure of the interfacial tension. The design of a cell mimetic environment with nano-scale confinement has also been demonstrated with diffusion measurements on macromolecules (PEG and Ficoll70) within droplets that are further structured internally using agarose gel networks. Containing the agarose gel in droplets appears to provide very reproducible and homogeneous network environments, enabling quantitative agreement of Ficoll70 dynamics with a theoretical model, with no fit parameters, and, with PEG, yielding a systematic polymer-size dependent slowing down in the network. This is in contrast with bulk agarose, where identical macromolecular diffusion measurements indicate the presence of heterogeneities with water pockets.

Tunable hydrodynamics: a field-frequency phase diagram of a non-equilibrium order-to-disorder transition.

We present experiments on a model system consisting of dielectric (silicone oil) drops in a "leaky dielectric" (castor oil) carrier fluid that exhibits dynamic non-equilibrium phases as a function of the amplitude and frequency of an external AC electric field. At high frequencies, the dielectric drops are pinned to a periodic lattice by dielectrophoretic forces induced by a patterned bottom electrode. Beginning with this state of imposed order, we examine the processes that take this system from order to disorder, with decreasing frequency corresponding to an increase in the range of the hydrodynamic forces. We find two kinds of disorder, shape- and translational disorder, that occur in frequency-amplitude space. We also find regimes where drop breakup is dominant, and where order/disorder of large drops can be probed without significant drop breakup. With decreasing frequency (i.e., increasing hydrodynamic coupling between drops) and on timescales from seconds to minutes, the drops exhibit motion that resembles Brownian motion of particles in a crystal, with an effective temperature that increases with the strength of the electrohydrodynamic driving force. In this limit, the system behaves like a thermal system and the lattice is seen to melt at an effective Lindemann parameter of Leff ∼ 0.08. This non-equilibrium thermodynamics, probed on timescales from seconds to minutes, likely arises from the pseudo-random velocity fields in the carrier fluid, as evidenced by the fractional, t3/2, super-diffusive tracer dynamics at shorter timescales.

The effect of crowder charge in a model polymer-colloid system for macromolecular crowding: Polymer structure and dynamics.

We have examined the effect of crowder particle charge on macromolecular structure, studied via small-angle neutron scattering, and translational dynamics, studied via pulsed-field gradient NMR, in addition to bulk viscosity measurements, in a polymer macromolecule (polyethylene glycol)-nanoparticle crowder (polysucrose, Ficoll70) model system, in the case where polymer size and crowder size are comparable. While there are modest effects of crowder charge on polymer dynamics at relatively low packing fractions, there is only a tiny effect at the high packing fractions that represent the limit of molecular crowding. We find, via different measures of macromolecular mobility, that the mobility of the flexible polymer in the crowding limit is 10-100 times larger than that of the compact, spherical crowder in spite of their similar size, implying that the flexible polymer chain is able to squeeze through crowder interstices.

Dynamics and cluster formation in charged and uncharged Ficoll70 solutions.

We apply pulsed-field-gradient NMR (PFG NMR) technique to measure the translational diffusion for both uncharged and charged polysaccharide (Ficoll70) in water. Analysis of the data indicates that the NMR signal attenuation above a certain packing fraction can be adequately fitted with a bi-exponential function. The self-diffusion measurements also show that the Ficoll70, an often-used compact, spherical polysucrose molecule, is itself nonideal, exhibiting signs of both softness and attractive interactions in the form of a stable suspension consisting of monomers and clusters. Further, we can quantify the fraction of monomers and clusters. This work strengthens the picture of the existence of a bound water layer within and around a porous Ficoll70 particle.

Contrasting the dynamics of elastic and non-elastic deformations across an experimental colloidal Martensitic transition.

We present a framework to segregate the roles of elastic and non-elastic deformations in the examination of real-space experiments of solid-solid Martensitic transitions. The Martensitic transformation of a body-centred-tetragonal (BCT) to a body-centred-orthorhombic (BCO) crystal structure has been studied in a model system of micron-scale ionic microgel colloids (P. S. Mohanty, P. Bagheri, S. Nöjd, A. Yethiraj and P. Schurtenberger, Phys. Rev. X, 2015, 5, 011030). Non-affine fluctuations, i.e., displacement fluctuations that do not arise from purely elastic (affine) deformations, are detected in particle configurations acquired from the experiment. Tracking these fluctuations serves as a highly sensitive tool in signaling the onset of the Martensitic transition and precisely locating particle rearrangements occurring at length scales of a few particle diameters. Particle rearrangements associated with non-affine displacement modes become increasingly favorable during the transformation process. The nature of the displacement fluctuation modes that govern the transformation are shown to be different from those predominant in an equilibrium crystal. We show that BCO crystallites formed through shear may, remarkably, co-exist with those resulting from local rearrangements within the same sample.

Frequency-Dependent Solvent Impedance and Colloid Microelectrophoresis Measurements in Partially Polar Solvents.

We carry out frequency-dependent solvent impedance measurements and alternating current (ac) colloid microelectrophoresis experiments in partially polar solvents in the low-frequency regime (0.25 Hz ≤ f ≤ 10 Hz). Solvent electrode polarization effects are quantified first in partially polar solvent mixtures containing bromocyclohexane (CHB). We find that the polarization capacitance from electrode polarization exhibits a clear power law behavior Cp = Cp0 f-m with power law exponent m = 0.25 ± 0.04. Once we account for electrode polarization effects, we are able to obtain quantitative mobilities in the low-frequency regime from our ac microelectrophoresis measurements; for these measurements, we use poly(methyl methacrylate colloids that are gravitationally confined to a plane while suspended in a low-polar solvent mixture of cis-trans decahydronapthalene and CHB. We find that the dimensionless electrophoretic mobility is constant, consistent with expectations for frequencies below the ion-diffusion frequency, and has a value E = 1.6 ± 0.4.

Quantifying disorder in colloidal films spin-coated onto patterned substrates.

Polycrystals of thin colloidal deposits, with thickness controlled by spin-coating speed, exhibit axial symmetry with local 4-fold and 6-fold symmetric structures, termed orientationally correlated polycrystals (OCPs). While spin-coating is a very facile technique for producing large-area colloidal deposits, the axial symmetry prevents us from achieving true long-range order. To obtain true long-range order, we break this axial symmetry by introducing a patterned surface topography and thus eliminate the OCP character. We then examine symmetry-independent methods to quantify order in these disordered colloidal deposits. We find that all the information in the bond-orientational order parameters is well captured by persistent homology analysis methods that only use the centers of the particles as input data. It is expected that these methods will prove useful in characterizing other disordered structures.

Combining Diffusion NMR and Small-Angle Neutron Scattering Enables Precise Measurements of Polymer Chain Compression in a Crowded Environment.

The effect of particles on the behavior of polymers in solution is important in a number of important phenomena such as the effect of "crowding" proteins in cells, colloid-polymer mixtures, and nanoparticle "fillers" in polymer solutions and melts. In this Letter, we study the effect of spherical inert nanoparticles (which we refer to as "crowders") on the diffusion coefficient and radius of gyration of polymers in solution using pulsed-field-gradient NMR and small-angle neutron scattering (SANS), respectively. The diffusion coefficients exhibit a plateau below a characteristic polymer concentration, which we identify as the overlap threshold concentration c^{⋆}. Above c^{⋆}, in a crossover region between the dilute and semidilute regimes, the (long-time) self-diffusion coefficients are found, universally, to decrease exponentially with polymer concentration at all crowder packing fractions, consistent with a structural basis for the long-time dynamics. The radius of gyration obtained from SANS in the crossover regime changes linearly with an increase in polymer concentration, and must be extrapolated to c^{⋆} in order to obtain the radius of gyration of an individual polymer chain. When the polymer radius of gyration and crowder size are comparable, the polymer size is very weakly affected by the presence of crowders, consistent with recent computer simulations. There is significant chain compression, however, when the crowder size is much smaller than the polymer radius gyration.

The effect of confinement on the electrohydrodynamic behavior of droplets in a microfluidic oil-in-oil emulsion.

A two-fluid emulsion (silicone oil drops in the "leaky dielectric", castor oil) with electrohydrodynamically driven flows can serve as a model system for tunable studies of hydrodynamic interactions [Varshney et al., Sci. Rep., 2012, 2, 738]. Flows on multiple length- and time-scales have been observed but the underlying mechanism for these chaotic, multi-scale flows is not understood. In this work, we conducted experiments varying the thickness of the test cell to examine the role of substrate interactions on size distribution, mean square displacement and velocity of the drops as a function of the electric field strength. We find that the electric capillary number, CaE, at the threshold of drop breakup is of order unity for cell thicknesses of 100 µm or thicker, but much larger for thinner cells. Above this threshold, there is a clear transition to super-diffusive droplet motions. In addition, we observe that there is a convective instability prior to the onset of chaotic flows, with the lengthscale associated with the convection rolls increasing linearly with an increase in the cell thickness. The fact that the convective instability appears to occur in the leaky dielectric castor oil regardless of whether the second component is liquid drops, solid particles, or dissolved dye has implications on the underlying mechanism for the unsteady flows.

Diffusion NMR studies of macromolecular complex formation, crowding and confinement in soft materials.

Label-free methods to obtain hydrodynamic size from diffusion measurements are desirable in environments that contain multiple macromolecular species at a high total concentration: one example is the crowded cellular environment. In complex, multi-species macromolecular environments - in this article, we feature aqueous systems involving polymers, surfactants and proteins - the link between dynamics and size is harder to unpack due to macromolecular crowding and confinement. In this review, we demonstrate that the pulsed-field gradient NMR technique, with its spectral separation of different chemical components, is ideal for studying the dynamics of the entire system simultaneously and without labelling, in a wide range of systems. The simultaneous measurement of the dynamics of multiple components allows for internal consistency checks and enables quantitative statements about the link between macromolecular dynamics, size, complex formation and crowding in soft materials.