Lymphocyte electrotaxis in vitro and in vivo.

Electric fields are generated in vivo in a variety of physiologic and pathologic settings, including penetrating injury to epithelial barriers. An applied electric field with strength within the physiologic range can induce directional cell migration (i.e., electrotaxis) of epithelial cells, endothelial cells, fibroblasts, and neutrophils suggesting a potential role in cell positioning during wound healing. In the present study, we investigated the ability of lymphocytes to respond to applied direct current (DC) electric fields. Using a modified Transwell assay and a simple microfluidic device, we show that human PBLs migrate toward the cathode in physiologically relevant DC electric fields. Additionally, electrical stimulation activates intracellular kinase signaling pathways shared with chemotactic stimuli. Finally, video microscopic tracing of GFP-tagged immunocytes in the skin of mouse ears reveals that motile cutaneous T cells actively migrate toward the cathode of an applied DC electric field. Lymphocyte positioning within tissues can thus be manipulated by externally applied electric fields, and may be influenced by endogenous electrical potential gradients as well.

Electrokinetics in nanochannels: part I. Electric double layer overlap and channel-to-well equilibrium.

In this paper a new model is described for calculating the electric potential field in a long, thin nanochannel with overlapped electric double layers. Electrolyte concentration in the nanochannel is predicted self-consistently via equilibrium between ionic solution in the wells and within the nanochannel. Differently than published models that require detailed iterative numerical solutions of coupled differential equations, the framework presented here is self-consistent and predictions are obtained solving a simple one-dimensional integral. The derivation clearly shows that the electric potential field depends on three new parameters: the ratio of ion density in the channel to ion density in the wells; the ratio of free-charge density to bulk ion density within the channel; and a modified Debye-Hückel thickness, which is the relevant scale for shielding of surface net charge. For completeness, three wall-surface boundary conditions are analyzed: specified zeta-potential; specified surface net charge density; and charge regulation. Predictions of experimentally observable quantities based on the model proposed here, such as depth-averaged electroosmotic flow and net ionic current, are significantly different than results from previous overlapped electric double layer models. In this first paper of a series of two, predictions are presented where channel depth is varied at constant well concentration. Results show that under conditions of electric double layer overlap, electroosmosis contributes only a small fraction of the net ionic current, and that most of the measurable current is due to ionic conduction in conditions of increased counterion density in the nanochannel. In the second of this two-paper series, predictions are presented where well-concentration is varied and the channel depth is held constant, and the model described here is employed to study the dependence of ion mobility on ionic strength, and compare predictions to measurements of ionic current as a function of channel depth and ion density.

Electrokinetics in nanochannels: part II. Mobility dependence on ion density and ionic current measurements.

In the first of this two-paper series, a new model was developed for calculating the electric potential field in a long, thin nanochannel with overlapped electric double layers. The model takes into account the dependence of ion mobility on local ion densities and pH. This model is used here to study and demonstrate the effect of ion density and pH on ionic current measurements. A comparison is shown of predictions based on each of three boundary conditions, as studied in Part I. The model developed in Part I is validated by comparing simulations with measurements of ionic current as a function of sodium borate concentration. Results show that predictions based on extended Debye-Hückel theory for ion mobility significantly improve the accuracy of simulations, but that these do not predict exact scaling behavior. A simple bulk conductivity measurement used as input parameter for the simulations, in place of the predicted bulk conductivity (K(0)), guarantees agreement with data in the thin EDL region. Results also indicate that the charge regulation boundary condition, complemented with an adequate bulk electrolyte model, provides better agreement with experimental trends than the specified zeta potential or specified surface net charge boundary conditions. Further, it is shown that currents due to advection (by electroosmotic flow) are in all cases studied less than 25% of the total current in the system.

Free-solution oligonucleotide separation in nanoscale channels.

In this paper, we report an experimental study of electrokinetic transport and separation of double-stranded deoxyribonucleic acid (dsDNA) oligonucleotides in custom-fabricated fused-silica nanochannels filled with a gel-free sodium borate aqueous buffer. Mixtures of fluorescently labeled dsDNA molecules in the range of 10-100 base pair (bp), fluorescein, and fluorescein-12-UTP (UTP) were separated in less than 120 s in channels of depth ranging from 40 to 1560 nm. We varied the channel depth and background buffer concentration to achieve a 0.006-0.2 range of Debye length-to-channel-half-depth ratio (lambdaD/h), and a 0.004-1.7 range of the ratio of length of dsDNA molecule to channel half-depth (l/h). We find observed oligonucleotide migration times depend on both l/h and lambdaD/h. Electrophoretic mobility estimates agree well with published (micrometer-scale channel) values for background electrolyte (BGE) concentrations greater than approximately 10 mM. At BGE concentrations of 1 and 5 mM, mobility estimates in our nanochannels are higher than published values. Of the cases studied, the highest separation sensitivities were achieved in 100 nm channels with 1-10 mM ion density buffers. Potential applications of this technology include rapid small-scale sequencing and other fluorescence-based oligonucleotide separation and detection assays.

Linear stability of a draining film squeezed between two approaching droplets.

In the present paper we analyze the effect of infinitesimal non-axisymmetric perturbations in determining the critical gap thickness at which a draining, finite radius thin-film becomes unstable. The film is part of the suspending fluid trapped between two approaching deformable drops under the action of a flow field. We carry out a linear stability analysis in the context of a quasi-static approximation where the rate of growth of the disturbances is assumed to be much faster than the rate of film drainage. An analytical solution is derived for the model in the special case of a uniformly thick film, for two types of perturbation: fixed-end and free-end. It is shown, for this special case, when the hydrodynamic force pushing the drops together from the external flow is constant, that the four most unstable disturbances are of the free-end kind, associated with the lowest frequency modes of azimuthal variation in the film thickness. Higher modes are stabilized by surface tension. Our analysis also shows that adopting the unretarded form of the van der Waals disjoining pressure yields results similar to the analysis when electromagnetic retardation effects are included in the calculation. A second case is analyzed where the film is also of uniform thickness but its lateral extent and the gap thickness are both time-dependent. This case was included to extend the predictions to glancing drop-collisions where the external hydrodynamic force is time-dependent. We find that there is a maximum capillary number below which the film becomes unstable, and that there is range of angles in the trajectory where the film becomes unstable, but that outside this range the film is stable.

Electrophoresis in nanochannels: brief review and speculation.

The relevant physical phenomena that dominate electrophoretic transport of ions and macromolecules within long, thin nanochannels are reviewed, and a few papers relevant to the discussion are cited. Sample ion transport through nanochannels is largely a function of their interaction with electric double layer. For small ions, this coupling includes the net effect of the external applied field, the internal field of the double layer, and the non-uniform velocity of the liquid. Adsorption/desorption kinetics and the effects of surface roughness may also be important in nanochannel electrophoresis. For macromolecules, the resulting motion is more complex as there is further coupling via steric interactions and perhaps polarization effects. These complex interactions and coupled physics represent a valuable opportunity for novel electrophoretic and chromatographic separations.

Two touching spherical drops in uniaxial extensional flow: analytic solution to the creeping flow problem.

We solve the problem of the creeping motion of a uniaxial extensional flow past two touching spherical drops when the line of centers is parallel to the axis of symmetry of the flow, using tangent sphere coordinates. We apply this solution to the case of two equal size drops. It provides an exact result for the equal and opposite force acting on each drop along the line of centers. We also use it to determine the magnitude of the internal recirculating flow in the vicinity of the rear stagnation point, which can be used to evaluate the importance of this flow on the film drainage process for two (nearly) touching drops in a coalescence process for the limiting case, Ca << 1.