Controlled rotation and vibration of patterned cell clusters using dielectrophoresis.

The localized motion of cells within a cluster is an important feature of living organisms and has been found to play roles in cell signaling, communication, and migration, thus affecting processes such as proliferation, transcription, and organogenesis. Current approaches for inducing dynamic movement into cells, however, focus predominantly on mechanical stimulation of single cells, affect cell integrity, and, more importantly, need a complementary mechanism to pattern cells. In this article, we demonstrate a new strategy for the mechanical stimulation of large cell clusters, taking advantage of dielectrophoresis. This strategy is based on the cellular spin resonance mechanism, but it utilizes coating agents, such as bovine serum albumin, to create consistent rotation and vibration of individual cells. The treatment of cells with coating agents intensifies the torque induced on the cells while reducing the friction at the cell-cell and cell-substrate interfaces, resulting in the consistent motion of the cells. Such localized motion can be modulated by varying the frequency and voltage of the applied sinusoidal AC signal and can be achieved in the absence and presence of flow. This strategy enables the survival and functioning of moving cells within large-scale clusters to be investigated.

An assessment of the dynamic stability of microorganisms on patterned surfaces in relation to biofouling control.

Microstructure-based patterned surfaces with antifouling capabilities against a wide range of organisms are yet to be optimised. Several studies have shown that microtopographic features affect the settlement and the early stages of biofilm formation of microorganisms. It is speculated that the fluctuating stress-strain rates developed on patterned surfaces disrupt the stability of microorganisms. This study investigated the dynamic interactions of a motile bacterium (Escherichia coli) with microtopographies in relation to initial settlement. The trajectories of E. coli across a patterned surface of a microwell array within a microchannel-based flow cell system were assessed experimentally with a time-lapse imaging module. The microwell array was composed of 256 circular wells, each with diameter 10 µm, spacing 7 µm and depth 5 µm. The dynamics of E. coli over microwell-based patterned surfaces were compared with those over plain surfaces and an increased velocity of cell bodies was observed in the case of patterned surfaces. The experimental results were further verified and supported by computational fluid dynamic simulations. Finally, it was stated that the nature of solid boundaries and the associated microfluidic conditions play key roles in determining the dynamic stability of motile bacteria in the close vicinity over surfaces.

Modifying dielectrophoretic response of nonviable yeast cells by ionic surfactant treatment.

Nonviable cells are essential biosystems, due to the functionalities they offer and their effects on viable cells. Therefore, the separation and immobilization of nonviable cells separately or in the vicinity of viable cells is of great importance for many fundamentals investigations in cell biology. However, most nonviable cells become less polarizable than the surrounding medium at conductivities above 0.01 S/m. This means that in such a medium, dielectrophoresis, despite its great versatilities for manipulation of cells, cannot be employed for immobilizing nonviable cells. Here, we present a novel approach to change the dielectrophoretic (DEP) response of nonviable yeast cells by treating them with low concentrations of ionic surfactants such as sodium dodecyl sulfate. After this treatment, they exhibit a strong positive DEP response, even at high medium conductivities. The capability of this treatment is demonstrated in two proof-of-concept experiments. First, we show the sorting and immobilization of viable and nonviable yeast cells, along consecutive microelectrode arrays. Second, we demonstrate the immobilization of viable and nonviable cells in the vicinity of each other along the same microelectrode array. The proposed technique allows DEP platforms to be utilized for the immobilization and subsequent postanalysis of both viable and nonviable cells with and without the presence of each other.

Dielectrophoresis with 3D microelectrodes fabricated by surface tension assisted lithography.

This paper demonstrates the utilization of 3D semispherical shaped microelectrodes for dielectrophoretic manipulation of yeast cells. The semispherical microelectrodes are capable of producing strong electric field gradients, and in turn dielectrophoretic forces across a large area of channel cross-section. The semispherical shape of microelectrodes avoids the formation of undesired sharp electric fields along the structure and also minimizes the disturbance of the streamlines of nearby passing fluid. The advantage of semispherical microelectrodes over the planar microelectrodes is demonstrated in a series of numerical simulations and proof-of-concept experiments aimed toward immobilization of viable yeast cells.

A novel approach to determine the efficacy of patterned surfaces for biofouling control in relation to its microfluidic environment.

Biofouling, the unwanted growth of sessile microorganisms on submerged surfaces, presents a serious problem for underwater structures. While biofouling can be controlled to various degrees with different microstructure-based patterned surfaces, understanding of the underlying mechanism is still imprecise. Researchers have long speculated that microtopographies might influence near-surface microfluidic conditions, thus microhydrodynamically preventing the settlement of microorganisms. It is therefore very important to identify the microfluidic environment developed on patterned surfaces and its relation with the antifouling behaviour of those surfaces. This study considered the wall shear stress distribution pattern as a significant aspect of this microfluidic environment. In this study, patterned surfaces with microwell arrays were assessed experimentally with a real-time biofilm development monitoring system using a novel microchannel-based flow cell reactor. Finally, computational fluid dynamics simulations were carried out to show how the microfluidic conditions were affecting the initial settlement of microorganisms.

Dynamic manipulation of modes in an optical waveguide using dielectrophoresis.

The emergence of optofluidics has brought a high degree of tuneability and reconfigurability to optical devices. These possibilities are provided by characteristics of fluids including mobility, wide range of index modulation, and abrupt interfaces that can be easily reshaped. In this work, we created a new class of optofluidic waveguides, in which suspended mesoparticles were employed to greatly enhance the flexibility of the system. We demonstrated tuneable quasi single mode waveguides using spatially controllable mesoparticles in optofluidics. The coupling of waveguiding modes into the assembly of mesoparticles produces strong interactions and resonant conditions, which promote the transitions of the waveguiding modes. The modal response of the system depends on the distribution of packed particles above the polymeric rib waveguide which can be readily controlled under the appropriate combination of dielectrophoresis and hydrodynamic forces.

Analysing calcium signalling of cells under high shear flows using discontinuous dielectrophoresis.

Immobilisation of cells is an important feature of many cellular assays, as it enables the physical/chemical stimulation of cells; whilst, monitoring cellular processes using microscopic techniques. Current approaches for immobilising cells, however, are hampered by time-consuming processes, the need for specific antibodies or coatings, and adverse effects on cell integrity. Here, we present a dielectrophoresis-based approach for the robust immobilisation of cells, and analysis of their responses under high shear flows. This approach is quick and label-free, and more importantly, minimises the adverse effects of electric field on the cell integrity, by activating the field for a short duration of 120 s, just long enough to immobilise the cells, after which cell culture media (such as HEPES) is flushed through the platform. In optimal conditions, at least 90% of the cells remained stably immobilised, when exposed to a shear stress of 63 dyn/cm(2). This approach was used to examine the shear-induced calcium signalling of HEK-293 cells expressing a mechanosensitive ion channel, transient receptor potential vaniloid type 4 (TRPV4), when exposed to the full physiological range of shear stress.

An investigation on platelet transport during thrombus formation at micro-scale stenosis.

This paper reports on an investigation of mass transport of blood cells at micro-scale stenosis where local strain-rate micro-gradients trigger platelet aggregation. Using a microfluidic flow focusing platform we investigate the blood flow streams that principally contribute to platelet aggregation under shear micro-gradient conditions. We demonstrate that relatively thin surface streams located at the channel wall are the primary contributor of platelets to the developing aggregate under shear gradient conditions. Furthermore we delineate a role for red blood cell hydrodynamic lift forces in driving enhanced advection of platelets to the stenosis wall and surface of developing aggregates. We show that this novel microfluidic platform can be effectively used to study the role of mass transport phenomena driving platelet recruitment and aggregate formation and believe that this approach will lead to a greater understanding of the mechanisms underlying shear-gradient dependent discoid platelet aggregation in the context of cardiovascular diseases such as acute coronary syndromes and ischemic stroke.