John R. de Bruyn

Drag force on a sphere in steady motion through a yield- stress fluid

We have studied the motion of spheres falling through yield-stress Carbopol gels. We measured the velocity of the falling sphere as a function of time and sphere density. Reproducible results were obtained when the experimental fluids were carefully prepared and homogenized. Three regimes of motion were observed. Spheres of high enough density reached a constant terminal velocity, as in Newtonian fluids. Below a critical density, the sphere came to a complete stop, while in an intermediate regime, the sphere continued to move but with a velocity which steadily decreased with time. We have also carefully characterized the rheological behavior of the fluids. The flow regimes observed for the falling sphere are analogous to those observed in creep tests for different applied stress levels. The yielding criterion and the drag force on the sphere obtained from our data are in excellent agreement with the longstanding but previously unconfirmed theoretical predictions of Beris et al. [J. Fluid Mech. 158, 219–24...

Apparatus for the study of Rayleigh–Bénard convection in gases under pressure

We review the history of experimental work on Rayleigh–Benard convection in gases, and then describe a modern apparatus that has been used in our experiments on gas convection. This system allows for the study of patterns in a cell with an aspect ratio (cell radius/fluid layer depth) as large as 100, with the cell thickness uniform to a fraction of a μm, and with the pressure controlled at the level of one part in 105. This level of control can yield a stability of the critical temperature difference for the convective onset of better than one part in 104. The convection patterns are visualized and the temperature field can be inferred using the shadowgraph technique. We describe the flow visualization and image processing necessary for this. Some interesting results obtained with the system are briefly summarized.

Fingering instability of a gravitationally driven contact line

Results of an experimental study of the flow of a viscous fluid down an inclined plane are presented. The contact line at the front of the flow is straight in the early stages of the flow, then becomes unstable to the formation of fingers. From measurements of the contact line position as a function of time for angles of inclination α in the range 0°<α≤32°, the flow is analyzed before and after the instability occurs, and the development of the finger pattern is parametrized.

Scaling and mesostructure of Carbopol dispersions

Rheological measurements were performed on aqueous dispersions of two commercial crosslinked polymer microgels, Carbopol Ultrez 10 and Carbopol ETD 2050, prepared over a wide range of concentration and pH. For all concentrations studied, both the yield stress and the elastic modulus initially increased dramatically with pH and displayed broad peaks at intermediate pH. This is consistent with the onset of jamming of the Carbopol particles due to a rapid increase in particle size caused by osmotic swelling in the presence of NaOH. Scaling of both yield stress and elasticity with concentration was observed only at higher concentrations, which we believe indicates a change from a percolated structure at low volume fractions to a space filling network of compressed particles at high volume fractions. This model is supported by confocal microscopy of fluorescently dyed Carbopol dispersions.

Dynamic light scattering study of inhibition of nucleation and growth of hydroxyapatite crystals by osteopontin.

We study the effect of isoforms of osteopontin (OPN) on the nucleation and growth of crystals from a supersaturated solution of calcium and phosphate ions. Dynamic light scattering is used to monitor the size of the precipitating particles and to provide information about their concentration. At the ion concentrations studied, immediate precipitation was observed in control experiments with no osteopontin in the solution, and the size of the precipitating particles increased steadily with time. The precipitate was identified as hydroxyapatite by X-ray diffraction. Addition of native osteopontin (nOPN) extracted from rat bone caused a delay in the onset of precipitation and reduced the number of particles that formed, but the few particles that did form grew to a larger size than in the absence of the protein. Recombinant osteopontin (rOPN), which lacks phosphorylation, caused no delay in initial calcium phosphate precipitation but severely slowed crystal growth, suggesting that rOPN inhibits growth but not nucleation. rOPN treated with protein kinase CK2 to phosphorylate the molecule (p-rOPN) produced an effect similar to that of nOPN, but at higher protein concentrations and to a lesser extent. These results suggest that phosphorylations are critical to OPN’s ability to inhibit nucleation, whereas the growth of the hydroxyapatite crystals is effectively controlled by the highly acidic OPN polypeptide. This work also demonstrates that dynamic light scattering can be a powerful tool for delineating the mechanism of protein modulation of mineral formation.

Peptides of Matrix Gla Protein Inhibit Nucleation and Growth of Hydroxyapatite and Calcium Oxalate Monohydrate Crystals

Matrix Gla protein (MGP) is a phosphorylated and γ-carboxylated protein that has been shown to prevent the deposition of hydroxyapatite crystals in the walls of blood vessels. MGP is also expressed in kidney and may inhibit the formation of kidney stones, which mainly consist of another crystalline phase, calcium oxalate monohydrate. To determine the mechanism by which MGP prevents soft-tissue calcification, we have synthesized peptides corresponding to the phosphorylated and γ-carboxylated sequences of human MGP in both post-translationally modified and non-modified forms. The effects of these peptides on hydroxyapatite formation and calcium oxalate crystallization were quantified using dynamic light scattering and scanning electron microscopy, respectively. Peptides YGlapS (MGP1-14: YγEpSHEpSMEpSYELNP), YEpS (YEpSHEpSMEpSYELNP), YGlaS (YγESHESMESYELNP) and SK-Gla (MGP43-56: SKPVHγELNRγEACDD) inhibited formation of hydroxyapatite in order of potency YGlapS > YEpS > YGlaS > SK-Gla. The effects of YGlapS, YEpS and YGlaS on hydroxyapatite formation were on both crystal nucleation and growth; the effect of SK-Gla was on nucleation. YGlapS and YEpS significantly inhibited the growth of calcium oxalate monohydrate crystals, while simultaneously promoting the formation of calcium oxalate dihydrate. The effects of these phosphopeptides on calcium oxalate monohydrate formation were on growth of crystals rather than nucleation. We have shown that the use of dynamic light scattering allows inhibitors of hydroxyapatite nucleation and growth to be distinguished. We have also demonstrated for the first time that MGP peptides inhibit the formation of calcium oxalate monohydrate. Based on the latter finding, we propose that MGP function not only to prevent blood-vessel calcification but also to inhibit stone formation in kidney.

Frequency-dependent viscoelasticity measurement by atomic force microscopy

We demonstrate a new technique for investigating viscoelastic properties of soft materials using the atomic force microscope. A small oscillatory voltage is added to the deflection signal of the atomic force microscope causing a vertical oscillatory sample motion. Monitoring the amplitude and phase of this motion allows determination of the viscous and elastic moduli of the sample as a function of frequency during contact imaging. This technique is applied to suspended poly(vinyl alcohol) nanofibers and poly(vinyl alcohol) hydrogels, giving results similar to those measured using traditional static methods. However, the moduli of both the fibers and the hydrogels show a significant frequency dependence. The Youngs modulus of the fibers increases with frequency, while for the viscoelastic hydrogels, the storage modulus dominates the mechanical response at low frequency whereas the loss modulus dominates at high frequency.

Experiments on three systems with non-variational aspects

Abstract We present recent experimental results for three pattern-forming systems in which non-variational effects play an important role. The first is thermal convection in a shallow horizontal layer of fluid with temperature-dependent properties. In this system, a hexagonal lattice of convection cells forms at onset. This lattice becomes unstable to rolls when the temperature difference is increased sufficiently. In the “roll” state, the roll are curved and the system forms stable rotating spirals. The rotating spiral states are associated with non-variational effects. Secondly, we discuss the formation of localized pulses in binary-mixture convection near onset. These pulses would not exist in a potential system. In narrow channels, they have been observed as stable states. In systems which are spatially extended in two dimensions they can form spontaneously, and can be long-lived. The third topic which we discuss is the Kuppers-Lortz instability in a thin horizontal layer of a Boussinesq fluid heated from below and rotated about a vertical axis. In this case, the pattern which forms immediately above the onset of convection is non-periodically time dependent even though the amplitude grows continuously from zero as the temperature difference is increased. The dominant mechanism of the instability is found to involve the motion of boundaries between coherent regions of convection rolls of a given orientation. The time dependence could not occur in a variational system. Since it occurs for arbitrarily small amplitudes, one might hope that it is amenable to theoretical analysis.

MIiSR: Molecular Interactions in Super-Resolution Imaging Enables the Analysis of Protein Interactions, Dynamics and Formation of Multi-protein Structures.

Our current understanding of the molecular mechanisms which regulate cellular processes such as vesicular trafficking has been enabled by conventional biochemical and microscopy techniques. However, these methods often obscure the heterogeneity of the cellular environment, thus precluding a quantitative assessment of the molecular interactions regulating these processes. Herein, we present Molecular Interactions in Super Resolution (MIiSR) software which provides quantitative analysis tools for use with super-resolution images. MIiSR combines multiple tools for analyzing intermolecular interactions, molecular clustering and image segmentation. These tools enable quantification, in the native environment of the cell, of molecular interactions and the formation of higher-order molecular complexes. The capabilities and limitations of these analytical tools are demonstrated using both modeled data and examples derived from the vesicular trafficking system, thereby providing an established and validated experimental workflow capable of quantitatively assessing molecular interactions and molecular complex formation within the heterogeneous environment of the cell.

Crossover between surface tension and gravity-driven instabilities of a thin fluid layer on a horizontal cylinder

A thin annular layer of fluid coating a cylinder is subject to two different instabilities. One, driven by surface tension, is analogous to the Rayleigh instability of a liquid jet. The other is the Rayleigh–Taylor instability, which is driven by gravity. Measurements of the wavelength and growth rate of periodic patterns of droplets which develop as a result of the instability of such a fluid layer are reported for cylinders with radius r in the range 0.0011<r<1.27 cm. For small r the wavelength and growth rate of the pattern are in agreement with theoretical predictions for the surface-tension-driven instability. For large r, the Rayleigh–Taylor instability is observed. At intermediate r there is a region of crossover between the two limiting cases.