Argon and oxygen plasma treatment increases hydrophilicity and reduces adhesion of silicon-incorporated diamond-like coatings.

In this study, the structure, adhesion, and cell viability characteristics of silicon-incorporated diamond-like carbon (Si-DLC) coatings on fused silica substrates were investigated. The effects of argon and oxygen postprocessing plasma treatments on the Si-DLC coatings were also studied. The contact angle results showed that the Si-DLC coatings were more hydrophilic than the uncoated surfaces, and postprocessing plasma treatment increased the hydrophilicity of the Si-DLC coatings. Atomic force microscopy and profilometry confirmed that postprocessing plasma treatment increased the thickness and roughness of the Si-DLC coatings. The results of microscratch testing indicated that the plasma treatments reduced the adhesion of the coatings. The x-ray photoelectron spectroscopy (XPS) showed the presence of carbon, oxygen, and silicon in the Si-DLC coatings before and after the plasma treatments. These results show that the postprocessing plasma treatment significantly reduced the atomic percentage of the carbon in the Si-DLC coatings. XPS also confirmed the presence of carbon in the form of sp3(C-C), sp2(C=C), C-O, and C=O bonds in the Si-DLC coatings; it showed that postprocessing treatments significantly increased the percentage of oxygen in the Si-DLC coatings. Fourier transform infrared spectroscopy (FTIR) analysis showed features associated with C-OH stretching, C-H bending, as well as Si-CH2 and C-H bending in the Si-DLC coating. The XPS and FTIR results confirmed that the plasma treatment caused dissociation of the sp2 and sp3 bonds and formation of C-OH bonds. The contact angle data indicated that postprocessing treatment increased the hydrophilicity of the Si-DLC coating. Similar to the uncoated substrates, L929 cells showed no change in cell viability when cultured on Si-DLC coatings. These results of the study indicate the suitability of Si-DLC coatings as inert coatings for medical and biotechnology applications.

DNA looping by two 5-methylcytosine-binding proteins quantified using nanofluidic devices.

BACKGROUND: MeCP2 and MBD2 are members of a family of proteins that possess a domain that selectively binds 5-methylcytosine in a CpG context. Members of the family interact with other proteins to modulate DNA packing. Stretching of DNA-protein complexes in nanofluidic channels with a cross-section of a few persistence lengths allows us to probe the degree of compaction by proteins. RESULTS: We demonstrate DNA compaction by MeCP2 while MBD2 does not affect DNA configuration. By using atomic force microscopy (AFM), we determined that the mechanism for compaction by MeCP2 is the formation of bridges between distant DNA stretches and the formation of loops. CONCLUSIONS: Despite sharing a similar specific DNA-binding domain, the impact of full-length 5-methylcytosine-binding proteins can vary drastically between strong compaction of DNA and no discernable large-scale impact of protein binding. We demonstrate that ATTO 565-labeled MBD2 is a good candidate as a staining agent for epigenetic mapping.

Physicochemical parameters that underlie inkjet printing for medical applications.

One of the most common types of 3D printing technologies is inkjet printing due to its numerous advantages, including low cost, programmability, high resolution, throughput, and speed. Inkjet printers are also capable of fabricating artificial tissues with physiological characteristics similar to those of living tissues. These artificial tissues are used for disease modeling, drug discovery, drug screening, and replacements for diseased or damaged tissues. This paper reviews recent advancements in one of the most common 3D printing technologies, inkjet dispensing. We briefly consider common printing techniques, including fused deposition modeling (FDM), stereolithography (STL), and inkjet printing. We briefly discuss various steps in inkjet printing, including droplet generation, droplet ejection, interaction of droplets on substrates, drying, and solidification. We also discuss various parameters that affect the printing process, including ink properties (e.g., viscosity and surface tension), physical parameters (e.g., internal diameter of printheads), and actuation mechanisms (e.g., piezoelectric actuation and thermal actuation). Through better understanding of common 3D printing technologies and the parameters that influence the printing processes, new types of artificial tissues, disease models, and structures for drug discovery and drug screening may be prepared. This review considers future directions in inkjet printing research that are focused on enhancing the resolution, printability, and uniformity of printed structures.

Direct observation of confinement-induced diffusophoresis.

Nanofluidic devices have channel dimensions which come to within one order of magnitude of the Debye length of common aqueous solutions. Conventionally, external driving is used to create concentration polarization of ions and biomolecules in nanofluidic devices. Here we show that long-range ionic strength gradients intrinsic to all nanofluidic devices, even at equilibrium, also drive a drift of macromolecules. To demonstrate the effect, we confine long DNA to straight nanochannels of constant, rectangular cross-section (100 × 100 nm2) which are connected to large microfluidic reservoirs. The motion of DNA is observed in absence of any driving. We find that at low ionic strengths, molecules in nanochannels migrate toward the nano-micro interface, while they are undergoing purely diffusive motion in high salt. Using numerical models, we demonstrate that the motion is consistent with the ionic strength gradient at the micro-nano interface even at equilibrium, and that the dominant cause of the drift is diffusophoresis.

An improved hybrid continuum-atomistic four-way coupled model for electrokinetics in nanofluidics.

In this study, an efficient hybrid continuum-atomistic method is proposed to study electrokinetic transport of aqueous solutions in nanofluidics. The aqueous phase is considered as a continuous phase containing immersed ion particles. The behavior of the system is then simulated through utilization of an improved hybrid continuum-atomistic four-way coupled approach, including the MultiPhase Particle-In-Cell method for the short-ranged interaction between the ion particles, the Brownian force for the collision between the aqueous phase molecules and the ion particles, and a wall force accounting for the short-ranged interaction of ions and walls. The validation of the proposed model with the results of Molecular Dynamics simulations suggests that this model can be a promising approach for studying the electrokinetic phenomena in more complicated geometries where the Molecular Dynamics approach is computationally prohibitive. Finally, the effects of electrokinetic parameters, such as the height of the channel, the external electric field, and bulk ionic concentration, on the electroosmotic flow in a nanochannel are investigated and discussed.

Motor-like DNA motion due to an ATP-hydrolyzing protein under nanoconfinement.

We report that long double-stranded DNA confined to quasi-1D nanochannels undergoes superdiffusive motion under the action of the enzyme T4 DNA ligase in the presence of necessary co-factors. Inside the confined environment of the nanochannel, double-stranded DNA molecules stretch out due to self-avoiding interactions. In absence of a catalytically active enzyme, we see classical diffusion of the center of mass. However, cooperative interactions of proteins with the DNA can lead to directed motion of DNA molecules inside the nanochannel. Here we show directed motion in this configuration for three different proteins (T4 DNA ligase, MutS, E. coli DNA ligase) in the presence of their energetic co-factors (ATP, NAD+).

Theoretical Study of Molecular Transport Through a Permeabilized Cell Membrane in a Microchannel.

A two-dimensional model is developed to study the molecular transport into an immersed cell in a microchannel and to investigate the effects of finite boundary (a cell is suspended in a microchannel), amplitude of electric pulse, and geometrical parameter (microchannel height and size of electrodes) on cell uptake. Embedded electrodes on the walls of the microchannel generate the required electric pulse to permeabilize the cell membrane, pass the ions through the membrane, and transport them into the cell. The shape of electric pulses is square with the time span of 6 ms; their intensities are in the range of 2.2, 2.4, 2.6, 3 V. Numerical simulations have been performed to comprehensively investigate the molecular uptake into the cell. The obtained results of the current study demonstrate that calcium ions enter the cell from the anodic side (the side near positive electrode); after a while, the cell faces depletion of the calcium ions on a positive electrode-facing side within the microchannel; the duration of depletion depends on the amplitude of electric pulse and geometry that lasts from microseconds to milliseconds. By keeping geometrical parameters and time span constant, increment of a pulse intensity enhances molecular uptake and rate of propagation inside the cell. If a ratio of electrode size to cell diameter is larger than 1, the transported amount of Ca 2+ into the cell, as well as the rate of propagation, will be significantly increased. By increasing the height of the microchannel, the rate of uptake is decreased. In an infinite domain, the peak concentration becomes constant after reaching the maximum value; this value depends on the intra-extracellular conductivity and diffusion coefficient of interior and exterior domains of the cell. In comparison, the maximum concentration is changed by geometrical parameters in the microchannel. After reaching the maximum value, the peak concentration reduces due to the depletion of Ca 2+ ions within the microchannel. Electrophoretic velocity has a significant effect on the cell uptake.

Hybrid continuum-atomistic approach to model electrokinetics in nanofluidics.

In this study, for the first time, a hybrid continuum-atomistic based model is proposed for electrokinetics, electroosmosis and electrophoresis, through nanochannels. Although continuum based methods are accurate enough to model fluid flow and electric potential in nanofluidics (in dimensions larger than 4 nm), ionic concentration is too low in nanochannels for the continuum assumption to be valid. On the other hand, the non-continuum based approaches are too time-consuming and therefore is limited to simple geometries, in practice. Here, to propose an efficient hybrid continuum-atomistic method of modelling the electrokinetics in nanochannels; the fluid flow and electric potential are computed based on continuum hypothesis coupled with an atomistic Lagrangian approach for the ionic transport. The results of the model are compared to and validated by the results of the molecular dynamics technique for a couple of case studies. Then, the influences of bulk ionic concentration, external electric field, size of nanochannel, and surface electric charge on the electrokinetic flow and ionic mass transfer are investigated, carefully. The hybrid continuum-atomistic method is a promising approach to model more complicated geometries and investigate more details of the electrokinetics in nanofluidics.

Numerical modeling of bi-polar (AC) pulse electroporation of single cell in microchannel to create nanopores on its membrane.

AC electroporation of a single cell in a microchannel was numerically studied. A 15 µm diameter cell was considered in a microchannel 25 µm in height and the influences of AC electric pulse on its membrane were numerically investigated. The cell was assumed to be suspended between two electroporative electrodes embedded on the walls of a microchannel. An amplitude and a time span of applied electric pulse were chosen to be 80 kV/m and 10 µs, respectively. For different frequency values (50, 100, 200, and 500 kHz), simulations were performed to show how the cell membrane was electroporated and the creation of nanopores. Obtained numerical results show that the most and the largest nanopores are created around poles of cell (nearest points of cell membrane to the electrodes). The numerical simulations also demonstrate that increased frequency will slightly decrease electroporated area of the cell membrane; additionally, growth of the created nanopores will be stabilized. It has also been proven that size and number of the created nanopores will be decreased by moving from the poles to the equator of the cell. There is almost no nanopore created in the vicinity of the equator. Frequency affects the rate of generation of nanopores. In case of AC electroporation, creation of nanopores has two phases that periodically repeat over time. In each period, the pore density sharply increases and then becomes constant. Enhancement of the frequency will result in decrease in time span of the periods. In each period, size of the created nanopores sharply increases and then slightly decreases. However, until the AC electric pulse is present, overall trends of creation and development of nanopores will be ascending. Variation of the size and number of created nanopores can be explained by considering time variation of transmembrane potential (difference of electric potential on two sides of cell membrane) which is clear in the results presented in this study.

A theoretical study of single-cell electroporation in a microchannel.

Electroporation of a single cell in a microchannel was studied. The effects of electrical (e.g., strength of the electric pulse) and geometrical (e.g., microchannel height, electrode size and position) parameters on cell membrane permeabilization were investigated. The electrodes were assumed to be embedded in the walls of the microchannel; the cell was suspended between these two electrodes. By keeping the electric pulse constant, increasing the microchannel height reduces the number and the radius of the biggest nanopores, as well as the electroporated area of the cell membrane. If the width of the electrodes is bigger than the cell diameter, the transmembrane potential will be centralized and have a sinusoidal distribution around the cell if nanopores are not generated. As the width of the electrode decreases and becomes smaller than the cell diameter, the local transmembrane potential decreases; in the nonelectroporative area, the transmembrane potential distribution deviates from the sinusoidal behavior; the induced transmembrane potential also concentrates around the poles of the cell membrane (the nearest points of the cell membrane to the electrodes). During cell membrane permeabilization, the biggest nanopores are initially created at the poles and then the nanopore population expands toward the equator. The number of the created nanopores reaches its maximal value within a few microseconds; further presence of the electric pulse may not influence the number and location of the created nanopores anymore but will develop the generated nanopores. Strengthening the electric pulse intensifies the size and number of the created nanopores as well as the electroporated area on the cell membrane.

Electroosmotic flow in a water column surrounded by an immiscible liquid.

In this paper, we conducted numerical simulation of the electroosmotic flow in a column of an aqueous solution surrounded by an immiscible liquid. While governing equations in this case are the same as that in the electroosmotic flow through a microchannel with solid walls, the main difference is the types of interfacial boundary conditions. The effects of electric double layer (EDL) and surface charge (SC) are considered to apply the most realistic model for the velocity boundary condition at the interface of the two fluids. Effects on the flow field of ς-potential and viscosity ratio of the two fluids were investigated. Similar to the electroosmotic flow in microchannels, an approximately flat velocity profile exists in the aqueous solution. In the immiscible fluid phase, the velocity decreases to zero from the interface toward the immiscible fluid phase. The velocity in both phases increases with ς-potential at the interface of the two fluids. The higher values of ς-potential also increase the slip velocity at the interface of the two fluids. For the same applied electric field and the same ς-potential at the interface of the two fluids, the more viscous immiscible fluid, the slower the system moves. The viscosity of the immiscible fluid phase also affects the flatness of the velocity profile in the aqueous solution.

Electrokinetic transport through the nanopores in cell membrane during electroporation.

In electroporation, applied electric field creates hydrophilic nanopores in a cell membrane that can serve as a pathway for inserting biological samples to the cell. It is highly desirable to understand the ionic transfer and fluid flow through the nanopores in order to control and improve the cell transfection. Because of submicron dimensions, conventional theories of electrokinetics may lose their applicability in such nanopores. In the current study, the Poisson-Nernst-Planck equations along with modified Navier-Stokes equations and the continuity equation are solved in order to find electric potential, fluid flow, and ionic concentration through the nanopores. The results show that the electric potential, velocity field, and ionic concentration vary with the size of the nanopores and are different through the nanopores located at the front and backside of the cell membrane. However, on a given side of the cell membrane, angular position of nanopores has fewer influences on liquid flow and ionic transfer. By increasing the radius of the nanopores, the averaged velocity and ionic concentration through the nanopores are increased. It is also shown that, in the presence of electric pulse, electrokinetic effects (electroosmosis and electrophoresis) have significant influences on ionic mass transfer through the nanopores, while the effect of diffusion on ionic mass flux is negligible in comparison with electrokinetics. Increasing the radius of the nanopores intensifies the effect of convection (electroosmosis) in comparison with electrophoresis on ionic flux.

Electrokinetic transport through nanochannels.

This article presents a numerical study of the electrokinetic transport phenomena (electroosmosis and electrophoresis) in a three-dimensional nanochannel with a circular cross-section. Due to the nanometer dimensions, the Boltzmann distribution of the ions is not valid in the nanochannels. Therefore, the conventional theories of electrokinetic flow through the microchannels such as Poisson-Boltzmann equation and Helmholtz-Smoluchowski slip velocity approach are no longer applicable. In the current study, a set of coupled partial differential equations including Poisson-Nernst-Plank equation, Navier-Stokes, and continuity equations is solved to find the electric potential field, ionic concentration field, and the velocity field in the three-dimensional nanochannel. The effects of surface electric charge and the radius of nanochannel on the electric potential, liquid flow, and ionic transport are investigated. Unlike the microchannels, the electric potential field, ionic concentration field, and velocity field are strongly size-dependent in nanochannels. The electric potential gradient along the nanochannel also depends on the surface electric charge of the nanochannel. More counter ions than the coions are transported through the nanochannel. The ionic concentration enrichment at the entrance and the exit of the nanochannel is completely evident from the simulation results. The study also shows that the flow velocity in the nanochannel is higher when the surface electric charge is stronger or the radius of the nanochannel is larger.

Numerical studies of continuous nutrient delivery for tumour spheroid culture in a microchannel by electrokinetically-induced pressure-driven flow.

Continuous nutrient delivery to cells by pressure-driven flow is desirable for cell culture in lab-on-a-chip devices. An innovative method is proposed to generate an induced pressure-driven flow by using an electrokinetically-driven pump in a H-shape microchannel. A three-dimensional numerical model is developed to study the effectiveness of the proposed mechanism. It is shown that the average velocity of the generated pressure-driven flow is linearly dependent on the applied voltage. Considering the culture of a multicellular tumour spheroid (MTS) in such a microfluidic system, numerical simulations based on EMT6/Ro tumour cells is performed to find the effects of the nutrient distribution (oxygen and glucose), bulk velocity and channel size on the cell growth. Using an empirical formula, the growth of the tumour cell is studied. For low nutrient concentrations and low speed flows, it is found that the MTS grows faster in larger channels. It is also shown that, for low nutrient concentrations, a higher bulk liquid velocity provide better environment for MTS to grow. For lower velocities, it is found that the local MTS growth along the flow direction deviates from the average growth.