Abstract: Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.
Abstract: A bio-sensing method, based on the plasmonic property of gold nano-islands, has been developed for detection of exosomes in a clinical setting. The position of the gold plasmon band in the UV-Visible spectrum depends on the size and shape of gold nanoparticles as well as on the surrounding environment. By adsorbing various chemical entities, or binding them, the gold plasmon band will shift toward longer wavelengths and the shift is proportional to the concentration. Exosomes transport cargoes of molecules and genetic materials to proximal and distal cells. Presently, the standard method for their isolation and quantification from body fluids is by ultracentrifugation, not a practical method to be implemented in a clinical setting. Thus, a versatile and cutting-edge platform is required to selectively detect and isolate exosomes for further analysis at clinical level. The new sensing protocol, instead of antibodies, makes use of a specially synthesized polypeptide (Vn96), to capture and quantify the exosomes from different media, by binding the heat shock proteins from exosomes. The protocol has been established and optimized by using a glass substrate, in order to facilitate the next stage, namely the transfer of the protocol to a microfluidic environment. After each step of the protocol, the UV-Vis spectrum was recorded and the position of gold Localized Surface Plasmon Resonance (LSPR) band was measured. The sensing process was modelled, taking into account the characteristics of the nano-island structure, prepared by thermal convection and annealing. The optimal molar ratios of the most important chemical entities, involved in the detection of exosomes were calculated as well. Indeed, it was found that the results of the sensing process depend on the two major steps: the molar ratios of streptavidin to biotin-PEG-Vn96 and, the final step, the capture of exosomes by the biotin-PEG-Vn96 complex. The microfluidic device designed for sensing of exosomes consists of a glass substrate, sealed by a PDMS layer that contains the channel and a collecting chamber. In the device, the solutions of linker, cross-linker, etc., are pumped over the gold nano-islands and an Ocean Optics spectrometer is used to measure the position of the Au plasmon band at each step of the sensing. The experiments have shown that the shift of the Au LSPR band is proportional to the concentration of exosomes and, thereby, exosomes can be accurately quantified. An important advantage of the method is the ability to discriminate between exosomes having different origins.
Abstract: A low-cost paper-based microfluidic device (PAD) for the multiplex electrochemical determination of glucose, uric acid, and dopamine in biological fluids was developed. Using wax printing, PAD containing a central zone, six channels, and six detection zones was fabricated, and the electrodes were printed on detection zones using pre-made electrodes template. For each analyte, two detection zones were used. The carbon working electrode was coated with chitosan-BSA (and enzymes for glucose and uric acid). To detect glucose and uric acid, enzymatic reactions were employed. These reactions involve enzyme-catalyzed redox reactions of the analytes and produce free electrons for electrochemical measurement. Calibration curves were linear (R² > 0.980) in the range of 0-80 mM for glucose, 0.09–0.9 mM for dopamine, and 0–50 mM for uric acid, respectively. Blood samples were successfully analyzed by the proposed method.
Abstract: This paper sets to demonstrate a modeling of electrokinetic mixing employing electroosmotic stationary and time-dependent microchannel using alternate zeta patches on the lower surface of the micromixer in a lab on chip microfluidic device. Electroosmotic flow is amplified using different 2D and 3D model designs with alternate and geometric zeta potential values such as 25, 50, and 100 mV, respectively, to achieve high concentration mixing in the electrokinetically-driven microfluidic system. The enhancement of electrokinetic mixing is studied using Finite Element Modeling, and simulation workflow is accomplished with defined integral steps. It can be observed that the presence of alternate zeta patches can help inducing microvortex flows inside the channel, which in turn can improve mixing efficiency. Fluid flow and concentration fields are simulated by solving Navier-Stokes equation (implying Helmholtz-Smoluchowski slip velocity boundary condition) and Convection-Diffusion equation. The effect of the magnitude of zeta potential, the number of alternate zeta patches, etc. are analysed thoroughly. 2D simulation reveals that there is a cumulative increase in concentration mixing, whereas 3D simulation differs slightly with low zeta potential as that of the 2D model within the T-shaped micromixer for concentration 1 mol/m3 and 0 mol/m3, respectively. Moreover, 2D model results were compared with those of 3D to indicate the importance of the 3D model in a microfluidic design process.
Abstract: To mimic the natural circumstances of cell growth in an organism, we present three-dimensional (3D) scaffolds fabricated by microfluidics for cultivation. This work investigates the cellular behaviors of rat cardiomyocytes in gelatin 3D scaffolds compared to those on 2D control, such as proliferation, viability and morphology. We found that the scaffolds may induce skeletal differentiation of H9c2 cells.
Abstract: In this paper, a simple microfluidic device for monitoring algal cell behavior is proposed. An array of algal microwells is fabricated by PDMS soft-lithography using X-ray LIGA mold, placed on a glass substrate. Two layers of replicated PDMS and substrate are attached by oxygen plasma bonding, creating a microchannel for the microfluidic system. Algal cell are loaded into the microfluidic device, which provides positive charge on the bottom surface of wells. Algal cells, which are negative charged, can be attracted to the bottom of the wells via electrostatic interaction. By varying the concentration of algal cells in the loading suspension, it is possible to obtain wells with a single cell. Liquid medium for cells monitoring are flown continuously over the wells, providing nutrient and waste exchange between the well and the main flow. This device could lead to the uncovering of the quantitative biology of the algae, which is a key to effective and extensive algal utilizations in the field of biotechnology, food industry and bioenergy research and developments.
Abstract: We developed an effective microfluidic device for photoreactions with low reflectance and good heat conductance. The performance of this microfluidic device was tested by carrying out a photoreactive synthesis of benzopinacol and acetone from benzophenone and 2-propanol. The yield reached 36% with an irradiation time of 469.2 s and was improved by more than 30% when compared to the values obtained by the batch method. Therefore, the microfluidic device was found to be effective for improving the yields of photoreactions.
Abstract: In this paper, design, fabrication and coupled
multifield analysis of hollow out-of-plane silicon microneedle array
with piezoelectrically actuated microfluidic device for transdermal
drug delivery (TDD) applications is presented. The fabrication
process of silicon microneedle array is first done by series of
combined isotropic and anisotropic etching processes using
inductively coupled plasma (ICP) etching technology. Then coupled
multifield analysis of MEMS based piezoelectrically actuated device
with integrated 2×2 silicon microneedle array is presented. To predict
the stress distribution and model fluid flow in coupled field analysis,
finite element (FE) and computational fluid dynamic (CFD) analysis
using ANSYS rather than analytical systems has been performed.
Static analysis and transient CFD analysis were performed to predict
the fluid flow through the microneedle array. The inlet pressure from
10 kPa to 150 kPa was considered for static CFD analysis. In the
lumen region fluid flow rate 3.2946 μL/min is obtained at 150 V for
2×2 microneedle array. In the present study the authors have
performed simulation of structural, piezoelectric and CFD analysis
on three dimensional model of the piezoelectrically actuated
mcirofluidic device integrated with 2×2 microneedle array.
Abstract: We have developed a microfluidic device system for the continuous producting of nanoparticles, and we have clarified the relationship between the mixing performance of reactors and the particle size. First, we evaluated the mixing performance of reactors by carring out the Villermaux–Dushman reaction and determined the experimental conditions for producing AgCl nanoparticles. Next, we produced AgCl nanoparticles and evaluated the mixing performance and the particle size. We found that as the mixing performance improves the size of produced particles decreases and the particle size distribution becomes sharper. We produced AgCl nanoparticles with a size of 86 nm using the microfluidic device that had the best mixing performance among the three reactors we tested in this study; the coefficient of variation (Cv) of the size distribution of the produced nanoparticles was 26.1%.