Dielectrophoretic Microfluidic Device for Separating Microparticles Based on Size with Sub-Micron Resolution.

This article details the mathematical model of a microfluidic device aimed at separating any binary heterogeneous sample of microparticles into two homogeneous samples based on size with sub-micron resolution. The device consists of two sections, where the upstream section is dedicated to focusing of microparticles, while the downstream section is dedicated to separation of the focused stream of microparticles into two samples based on size. Each section has multiple planar electrodes of finite size protruding into the microchannel from the top and bottom of each sidewall; each top electrode aligns with a bottom electrode and they form a pair leading to multiple pairs of electrodes on each side. The focusing section subjects all microparticles to repulsive dielectrophoretic force, from each set of the electrodes, to focus them next to one of the sidewalls. This separation section pushes the big microparticles toward the interior, away from the wall, of the microchannel using repulsive dielectrophoretic force, while the small microparticles move unaffected to achieve the desired degree of separation. The operating frequency of the set of electrodes in the separation section is maintained equal to the cross-over frequency of the small microparticles. The working of the device is demonstrated by separating a heterogeneous mixture consisting of polystyrene microparticles of different size (radii of 2 and 2.25 µm) into two homogeneous samples. The mathematical model is used for parametric study, and the performance is quantified in terms of separation efficiency and separation purity; the parameters considered include applied electric voltages, electrode dimensions, outlet widths, number of electrodes, and volumetric flowrate. The separation efficiencies and separation purities for both microparticles are 100% for low volumetric flow rates, a large number of electrode pairs, large electrode dimensions, and high differences between voltages in both sections.

Modeling a Dielectrophoretic Microfluidic Device with Vertical Interdigitated Transducer Electrodes for Separation of Microparticles Based on Size.

This article conceptualizes and mathematically models a dielectrophoretic microfluidic device with two sets of interdigitated transducer vertical electrodes for separation of a binary heterogeneous mixture of particles based on size; each set of electrodes is located on the sidewalls and independently controllable. To achieve separation in the proposed microfluidic device, the small microparticles are subjected to positive dielectrophoresis and the big microparticles do not experience dielectrophoresis. The mathematical model consists of equations describing the motion of each microparticle, fluid flow profile, and electric voltage and field profiles, and they are solved numerically. The equations of motion take into account the influence of phenomena, such as inertia, drag, dielectrophoresis, gravity, and buoyancy. The model is used for a parametric study to understand the influence of parameters on the performance of the microfluidic device. The parameters studied include applied electric voltages, electrode dimensions, volumetric flow rate, and number of electrodes. The separation efficiency of the big and small microparticles is found to be independent of and dependent on all parameters, respectively. On the other hand, the separation purity of the big and small microparticles is found to be dependent on and independent of all parameters, respectively. The mathematical model is useful in designing the proposed microfluidic device with the desired level of separation efficiency and separation purity.

Halotolerant Marine Rhizosphere-Competent Actinobacteria Promote Salicornia bigelovii Growth and Seed Production Using Seawater Irrigation.

Salicornia bigelovii is a promising halophytic cash crop that grows in seawater of the intertidal zone of the west-north coast of the UAE. This study assess plant growth promoting (PGP) capabilities of halotolerant actinobacteria isolated from rhizosphere of S. bigelovii to be used as biological inoculants on seawater-irrigated S. bigelovii plants. Under laboratory conditions, a total of 39 actinobacterial strains were isolated, of which 22 were tolerant to high salinity (up to 8% w/v NaCl). These strains were further screened for their abilities to colonize S. bigelovii roots in vitro; the most promising ones that produced indole-3-acetic acid, polyamines (PA) or 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase (ACCD) were selected for rhizosphere-competency under naturally competitive environment. Three outstanding rhizosphere-competent isolates, Streptomyces chartreusis (Sc), S. tritolerans (St), and S. rochei (Sr) producing auxins, PA and ACCD, respectively, were investigated individually and as consortium (Sc/St/Sr) to determine their effects on the performance of S. bigelovii in the greenhouse. Individual applications of strains on seawater-irrigated plants significantly enhanced shoot and root dry biomass by 32.3-56.5% and 42.3-71.9%, respectively, in comparison to non-inoculated plants (control). In addition, plants individually treated with Sc, St and Sr resulted in 46.1, 60.0, and 69.1% increase in seed yield, respectively, when compared to control plants. Thus, the synergetic combination of strains had greater effects on S. bigelovii biomass (62.2 and 77.9% increase in shoot and root dry biomass, respectively) and seed yield (79.7% increase), compared to the control treatment. Our results also showed significant (P < 0.05) increases in the levels of photosynthetic pigments, endogenous auxins and PA, but a reduction in the levels of ACC in tissues of plants inoculated with Sc/St/Sr. We conclude that the consortium of isolates was the most effective treatment on S. bigelovii growth; thus confirmed by principal component and correlation analyses. To this best of our knowledge, this is the first report about halotolerant rhizosphere-competent PGP actinobacteria thriving in saline soils that can potentially contribute to promoting growth and increasing yield of S. bigelovii. These halotolerant actinobacterial strains could potentially be exploited as biofertilizers to sustain crop production in arid coastal areas.

Electrical detection of blood cells in urine.

Available methods for detecting blood in the urine (hematuria) can be problematic since results can be influenced by many factors in patients and in the lab settings, resulting in false positive or false negative results. This necessitates the development of new, accurate and easy-access methods that save time and effort. This study demonstrates a label-free and accurate method for detecting the presence of red and white blood cells (RBCs and WBCs) in urine by measuring the changes in the dielectric properties of urine upon increasing concentrations of both cell types. The current method could detect changes in the electrical properties of fresh urine over a short time interval, making this method suitable for detecting changes that cannot be recognized by conventional methods. Correcting for these changes enabled the detection of a minimum cell concentration of 102 RBCs per ml which is not possible by conventional methods used in the labs except for the semi-quantitative method that can detect 50 RBCs per ml, but it is a lengthy and involved procedure, not suitable for high volume labs. This ability to detect very small amount of both types of cells makes the proposed technique an attractive tool for detecting hematuria, the presence of which is indicative of problems in the excretory system.

Graphene-based 3D scaffolds in tissue engineering: fabrication, applications, and future scope in liver tissue engineering.

Tissue engineering embraces the potential of recreating and replacing defective body parts by advancements in the medical field. Being a biocompatible nanomaterial with outstanding physical, chemical, optical, and biological properties, graphene-based materials were successfully employed in creating the perfect scaffold for a range of organs, starting from the skin through to the brain. Investigations on 2D and 3D tissue culture scaffolds incorporated with graphene or its derivatives have revealed the capability of this carbon material in mimicking in vivo environment. The porous morphology, great surface area, selective permeability of gases, excellent mechanical strength, good thermal and electrical conductivity, good optical properties, and biodegradability enable graphene materials to be the best component for scaffold engineering. Along with the apt microenvironment, this material was found to be efficient in differentiating stem cells into specific cell types. Furthermore, the scope of graphene nanomaterials in liver tissue engineering as a promising biomaterial is also discussed. This review critically looks into the unlimited potential of graphene-based nanomaterials in future tissue engineering and regenerative therapy.

Dielectrophoresis-based 3D-focusing of microscale entities in microfluidic devices.

This article presents a validated mathematical model of a dielectrophoresis (DEP)-based microfluidic device capable of 3D-focusing microscale entities at any lateral location inside the microchannel. The microfluidic device employs planar, independently controllable, interdigitated transducer (IDT) electrodes on either side of the microchannel. The developed model is used for understanding the influence of different geometric and operating parameters on 3D focusing, and it comprises of motion equation, Navier-Stokes equation, continuity equation, and electric potential equation (Laplace equation). The model accounts for forces associated with inertia, gravity, buoyancy, virtual mass, drag, and DEP. The model is solved using finite difference method. The findings of the study indicate that the 3D focusing possible with the proposed microfluidic device is independent of microscale entity's size and initial position, microchannel height, and volumetric flow rate. In contrast, 3D focusing achievable with the microfluidic device is dependent on the applied electric potential, protrusion width of electrodes, and width of electrode/gap. Additionally, the lateral position of 3D focused can be controlled by varying the applied electric potential. The advantage of the proposed microfluidic device is that it is simple to construct while capable of achieving 3D focusing at any lateral location inside the microchannel.

Bilirubin detoxification using different phytomaterials: characterization and in vitro studies.

BACKGROUND: Activated carbon (AC) is a common adsorbent that is used in both artificial and bioartificial liver devices. METHODS: Three natural materials - date pits of Phoenix dactylifera (fruit), Simmondsia chinensis (jojoba) seeds, and Scenedesmus spp. (microalgae) - were used in the present investigation as precursors for the synthesis of AC using physical activation. The chemical structures and morphology of AC were analyzed. Then, AC's bilirubin adsorption capacity and its cytotoxicity on normal liver (THLE2) and liver cancer (HepG2) cells were characterized. RESULTS: Compared with the other raw materials examined, date-pit AC was highly selective and showed the most effective capacity of bilirubin adsorption, as judged by isotherm-modeling analysis. MTT in vitro analysis indicated that date-pit AC had the least effect on the viability of both THLE2 and HepG2 cells compared to jojoba seeds and microalgae. All three biomaterials under investigation were used, along with collagen and Matrigel, to grow cells in 3D culture. Fluorescent microscopy confirmed date-pit AC as the best to preserve liver cell integrity. CONCLUSION: The findings of this study introduce date-pit-based AC as a novel alternative biomaterial for the removal of protein-bound toxins in bioartificial liver devices.

Analysis of dielectrophoresis based 3D-focusing in microfluidic devices with planar electrodes.

This article models a dielectrophoresis based approach for achieving 3D focusing, of micro-scale objects, in microfluidic devices. The microfluidic device employs four planar electrodes; two electrodes each on the top and bottom surface of the microchannel and each slightly protrude into the microchannel. Each electrode establishes electric field with the neighboring electrode on the same and opposite surfaces. The dielectrophoretic force pushes the micro-scale objects both the directions transverse to the flow direction to achieve the desired 3D focusing. The developed model accounts for various forces such as that associated with inertia, sedimentation, drag, and dielectrophoresis. Finite difference method is used for calculating the electric field and dielectrophoretic force as well as the displacements of micro-scale objects in the microchannel. Several geometric and operating parameters influence the trajectory of micro-scale objects. There exists a threshold voltage beyond which there is no increase in levitation height.

3D culture of mouse gastric stem cells using porous microcarriers.

The lining epithelium of the stomach includes multipotent stem cells which undergo proliferation and migration-associated differentiation. These cells give rise to multiple cell lineages that produce mucus, acid, pepsinogen and various hormones/peptides. A 3D culture for stem cells would facilitate identification of the factors that control proliferation and/or differentiation programs. Here, we report on the use of disk-like ImmobaSil HD silicone-rubber matrix based microcarriers that are permeable to oxygen and reduce the creation of toxic environment within the center of the microcarrierd for culturing the mouse gastric stem (mGS) cells. We define several parameters that affect the initial cell attachment such as size of cell inoculum, serum concentration, mode and speed of agitation. We show that although such a microcarrier allows for attachment and growth of gastric stem cells, it does not lend itself and does not support the functional differentiation of such cells.

Assessment of Tree Leaves Flakes Mixed with Crude Glycerol as a Bioenergy Source.

The gasification and combustion of dry tree leaves and the cogasification of dry tree leaves soaking crude glycerol were studied experimentally. An updraft fixed bed gasification and combustion system was built. The operation was conducted at different air to fuel ratios. Results show more stable combustion and more effective heat transfer to furnace walls for the cases when tree leaves flakes are mixed with 20 percent (on mass basis) of crude glycerol, as compared with the case when only dry tree leaves are used as fuel. TGA analysis was also conducted for the two fuels used under both air and nitrogen environments. For the crude glycerol, four phases of pyrolysis and gasification were noticed under either of the two surrounding gaseous media (air or nitrogen). For the dry tree leaves, the pyrolysis under nitrogen shows only a simple smooth pyrolysis and gasification curve without showing the different distinct phases that were otherwise identified when the pyrolysis is conducted under air environment. Moreover, the air TGA results lead to more gasification due to the char oxidation at high temperatures. DTG results are also presented and discussed.

RF microalgal lipid content characterization.

Most conventional techniques for the determination of microalgae lipid content are time consuming and in most cases are indirect and require excessive sample preparations. This work presents a new technique that utilizes radio frequency (RF) for rapid lipid quantification, without the need for sample preparation. Tests showed that a shift in the resonance frequency of a RF open-ended coaxial resonator and a gradual increase in its resonance magnitude may occur as the lipids content of microalgae cells increases. These response parameters can be then calibrated against actual cellular lipid contents and used for rapid determination of the cellular lipids. The average duration of lipid quantification using the proposed technique was of about 1 minute, which is significantly less than all other conventional techniques, and was achieved without the need for any time consuming treatment steps.

Effect of membranes on oxygen transfer rate and consumption within a newly developed three-compartment bioartificial liver device: Advanced experimental and theoretical studies.

A mathematical model is developed to predict oxygen transfer in the fiber-in-fiber (FIF) bioartificial liver device. The model parameters are taken from the constructed and tested FIF modules. We extended the Krogh cylinder model by including one more zone for oxygen transfer. Cellular oxygen uptake was based on Michaelis-Menten kinetics. The effect of varying a number of important model parameters is investigated, including (1) oxygen partial pressure at the inlet, (2) the hydraulic permeability of compartment B (cell region), (3) the hydraulic permeability of the inner membrane, and (4) the oxygen diffusivity of the outer membrane. The mathematical model is validated by comparing its output against the experimentally acquired values of an oxygen transfer rate and the hydrostatic pressure drop. Three governing simultaneous linear differential equations are derived to predict and validate the experimental measurements, e.g., the flow rate and the hydrostatic pressure drop. The model output simulated the experimental measurements to a high degree of accuracy. The model predictions show that the cells in the annulus can be oxygenated well even at high cell density or at a low level of gas phase PG if the value of the oxygen diffusion coefficient Dm is 16 × 10(-5) . The mathematical model also shows that the performance of the FIF improves by increasing the permeability of polypropylene membrane (inner fiber). Moreover, the model predicted that 60% of plasma has access to the cells in the annulus within the first 10% of the FIF bioreactor axial length for a specific polypropylene membrane permeability and can reach 95% within the first 30% of its axial length.

Experimental evaluation and theoretical modeling of oxygen transfer rate for the newly developed hollow fiber bioreactor with three compartments.

Bioartificial liver support provides a bridge to transplantation which is at present the only proven specific treatment for acute liver failure. In this paper, a novel multi-coaxial hollow fiber bioreactor so-called "Fibre-in-Fibre FIF Bioartificial liver device" with three compartments is experimentally and mathematically studied. The mathematical model in this paper is an extension of Krogh cylinder model for hollow fibre devices by including one more zone for oxygen transfer, i.e. oxygenation compartment. Three simultaneous linear differential equations were derived for pressure in plasma and cell compartments and flow rate in cell compartment. To validate the model, Oxygen Transfer Rate and hydrostatic pressure experimental measurements for different flow rates, 17-400 ml/min, and different number of hollow fibres pairs are used. Several important parameters of the Michaelis-Menten was investigated, namely, constant Vmax (the maximum oxygen consumption per unit volume of the cell mass), the oxygen partial pressure, the flow rate of the perfusate at device inlet. The results showed that the oxygenation compartment should easily secure Oxygen to the cells in compartment B.

Growth, metabolic activity, and productivity of immobilized and freely suspended CHO cells in perfusion culture.

Chinese hamster ovary (CHO) cells producing ß-galactosidase (ß-gal) were successfully cultured on silicone-based porous microcarriers (ImmobaSil FS) in a 1 L stirred-tank perfusion bioreactor. We studied the growth, metabolism, and productivity of free and immobilized cells to understand cellular activity in immobilized conditions. CHO cells attached to ImmobaSil FS significantly better than to other microcarriers. Scanning electron microscope images showed that the CHO cells thoroughly colonized the porous surfaces of the ImmobaSil FS, exhibiting a spherical morphology with microvilli that extended to anchorage cells on the silicone surface. In perfusion culture, the concentration of the attached cells reached 8 × 10(8) cells/mL of carrier, whereas those that remained freely suspended reached 2 × 10(7) cells/mL medium. The ß-gal concentration reached more than 5 unit/mL in perfusion culture, more than fivefold that of batch culture. The maximum concentration per microcarrier was proportional to the initial cell density. The specific growth rate, the specific ß-gal production rate, the percentage of S phase, and the oxygen uptake rate were all relatively lower for immobilized cells than freely suspended cells in the same bioreactor, indicating that not only do cells survive and grow to a greater extent in a free suspension state, but they are also metabolically more active than viable cells inside the pores of the microcarriers.