Search results for: microfluidic chips

Authors: Yana Posmitnaya, Galina Rudnitskaya, Tatyana Lukashenko, Anton Bukatin, Anatoly Evstrapov

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An important area of biological and medical research is the study of genetic mutations and polymorphisms that can alter gene function and cause inherited diseases and other diseases. The following methods to analyse DNA fragments are used: capillary electrophoresis and electrophoresis on microfluidic chip (MFC), mass spectrometry with electrophoresis on MFC, hybridization assay on microarray. Electrophoresis on MFC allows to analyse small volumes of samples with high speed and throughput. A soft lithography in polydimethylsiloxane (PDMS) was chosen for operative fabrication of MFCs. A master-form from silicon and photoresist SU-8 2025 (MicroChem Corp.) was created for the formation of micro-sized structures in PDMS. A universal topology which combines T-injector and simple cross was selected for the electrophoretic separation of the sample. Glass K8 and PDMS Sylgard® 184 (Dow Corning Corp.) were used for fabrication of MFCs. Electroosmotic flow (EOF) plays an important role in the electrophoretic separation of the sample. Therefore, the estimate of the quantity of EOF and the ways of its regulation are of interest for the development of the new methods of the electrophoretic separation of biomolecules. The following methods of surface modification were chosen to change EOF: high-frequency (13.56 MHz) plasma treatment in oxygen and argon at low pressure (1 mbar); 1% aqueous solution of polyvinyl alcohol; 3% aqueous solution of Kolliphor® P 188 (Sigma-Aldrich Corp.). The electroosmotic mobility was evaluated by the method of Huang X. et al., wherein the borate buffer was used. The influence of physical and chemical methods of treatment on the wetting properties of the PDMS surface was controlled by the sessile drop method. The most effective way of surface modification of MFCs, from the standpoint of obtaining the smallest value of the contact angle and the smallest value of the EOF, was the processing with aqueous solution of Kolliphor® P 188. This method of modification has been selected for the treatment of channels of MFCs, which are used for the separation of mixture of oligonucleotides fluorescently labeled with the length of chain with 10, 20, 30, 40 and 50 nucleotides. Electrophoresis was performed on the device MFAS-01 (IAI RAS, Russia) at the separation voltage of 1500 V. 6% solution of polydimethylacrylamide with the addition of 7M carbamide was used as the separation medium. The separation time of components of the mixture was determined from electropherograms. The time for untreated MFC was ~275 s, and for the ones treated with solution of Kolliphor® P 188 – ~ 220 s. Research of physical-chemical methods of surface modification of MFCs allowed to choose the most effective way for reducing EOF – the modification with aqueous solution of Kolliphor® P 188. In this case, the separation time of the mixture of oligonucleotides decreased about 20%. The further optimization of method of modification of channels of MFCs will allow decreasing the separation time of sample and increasing the throughput of analysis.

Keywords: electrophoresis, microfluidic chip, modification, nucleic acid, polydimethylsiloxane, soft lithography

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Authors: Ane Larrea, Victor Sebastian, Manuel Arruebo, Jesus Santamaria

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We present a gas-liquid microfluidic system as a reactor to obtain magnetite nanoparticles with an excellent degree of control regarding their crystalline phase, shape and size. Several types of microflow approaches were selected to prevent nanomaterial aggregation and to promote homogenous size distribution. The selected reactor consists of a mixer stage aided by ultrasound waves and a reaction stage using a N2-liquid segmented flow to prevent magnetite oxidation to non-magnetic phases. A milli-fluidic reactor was developed to increase the production rate where a magnetite throughput close to 450 mg/h in a continuous fashion was obtained.

Keywords: continuous production, magnetic nanoparticles, microfluidics, nanomaterials

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Authors: Adil Basuki Ahza, Michael Liong, Subarna Suryatman

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Healthy chips now dominating the snack market shelves. More than 80% processed snack foods in the market are chips. This research takes the advantages of twin extrusion technology to produce two types of product, i.e. directly expanded and intermediate ready-to-fry or microwavable chips. To improve the functional quality, the cereal-tuber based mix was enriched with antioxidant rich mix of temurui, celery, carrot and isolated soy protein (ISP) powder. Objectives of this research were to find best composite cassava-sorghum ratio, i.e. 60:40, 70:30 and 80:20, to optimize processing conditions of extrusion and study the microstructural, physical and sensorial characteristics of the final products. Optimization was firstly done by applying metering section of extruder barrel temperatures of 120, 130 and 140 °C with screw speeds of 150, 160 and 170 rpm to produce direct expanded product. The intermediate product was extruded in 100 °C and 100 rpm screw speed with feed moisture content of 35, 40 and 45%. The directly expanded products were analyzed for color, hardness, density, microstructure, and organoleptic properties. The results showed that interaction of ratio of cassava-sorghum and cooking methods affected the product's color, hardness, and bulk density (p<0.05). Extrusion processing conditions also significantly affected product's microstructure (p<0.05). The direct expanded snacks of 80:20 cassava-sorghum ratio and fried expanded one 70:30 and 80:20 ratio shown the best organoleptic score (slightly liked) while baking the intermediate product with microwave were resulted sensorial not acceptable quality chips.

Keywords: cassava-sorghum composite, extrusion, microstructure, physical characteristics

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Authors: Julia R. Beulig, Stefan Ohla, Detlev Belder

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In contrast to off-line analytical methods, lab-on-a-chip technology delivers direct information about the observed reaction. Therefore, microfluidic devices make an important scientific contribution, e.g. in the field of synthetic chemistry. Herein, the rapid generation of analytical data can be applied for the optimization of chemical reactions. These microfluidic devices enable a fast change of reaction conditions as well as a resource saving method of operation. In the presented work, we focus on the investigation of multiphase regimes, more specifically on a biphasic microfluidic droplet systems. Here, every single droplet is a reaction container with customized conditions. The biggest challenge is the rapid qualitative and quantitative readout of information as most detection techniques for droplet systems are non-specific, time-consuming or too slow. An exception is the electrospray mass spectrometry (ESI-MS). The combination of a reaction screening platform with a rapid and specific detection method is an important step in droplet-based microfluidics. In this work, we present a novel approach for synthesis optimization on the nanoliter scale with direct ESI-MS detection. The development of a droplet-based microfluidic device, which enables the modification of different parameters while simultaneously monitoring the effect on the reaction within a single run, is shown. By common soft- and photolithographic techniques a polydimethylsiloxane (PDMS) microfluidic chip with different functionalities is developed. As an interface for the MS detection, we use a steel capillary for ESI and improve the spray stability with a Teflon siphon tubing, which is inserted underneath the steel capillary. By optimizing the flow rates, it is possible to screen parameters of various reactions, this is exemplarity shown by a Domino Knoevenagel Hetero-Diels-Alder reaction. Different starting materials, catalyst concentrations and solvent compositions are investigated. Due to the high repetition rate of the droplet production, each set of reaction condition is examined hundreds of times. As a result, of the investigation, we receive possible reagents, the ideal water-methanol ratio of the solvent and the most effective catalyst concentration. The developed system can help to determine important information about the optimal parameters of a reaction within a short time. With this novel tool, we make an important step on the field of combining droplet-based microfluidics with organic reaction screening.

Keywords: droplet, mass spectrometry, microfluidics, organic reaction, screening

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Authors: Zara L. Sheady

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Fluidic oscillators are being utilised in an increasing number of applications in a wide variety of areas; these include on-board vehicle cleaning systems, flow separation control on aircraft and in fluidic circuitry. With this increased use, there is a further understanding required for the mechanics of the fluidics of the fluidic oscillator and why they work in the manner that they do. ANSYS CFX has been utilized to visualise the pressure and velocity within a microfluidic feedback oscillator. The images demonstrate how the pressure vortices build within the oscillator at the points where the velocity is diverted from linear motion through the oscillator. With an enhanced understanding of the pressure and velocity distributions within a fluidic oscillator, it will enable users of microfluidics to more greatly tailor fluidic nozzles to their specification.

Keywords: ANSYS CFX, control, fluidic oscillators, mechanics, pressure, relationship, velocity

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Authors: A. Aslihan Gokaltun, Martin L. Yarmush, Ayse Asatekin, O. Berk Usta

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In the last decade microfabrication processes including rapid prototyping techniques have advanced rapidly and achieved a fairly matured stage. These advances encouraged and enabled the use of microfluidic devices by a wider range of users with applications in biological separations, and cell and organoid cultures. Accordingly, a significant current challenge in the field is controlling biomolecular interactions at interfaces and the development of novel biomaterials to satisfy the unique needs of the biomedical applications. Poly(dimethylsiloxane) (PDMS) is by far the most preferred material in the fabrication of microfluidic devices. This can be attributed its favorable properties, including: (1) simple fabrication by replica molding, (2) good mechanical properties, (3) excellent optical transparency from 240 to 1100 nm, (4) biocompatibility and non-toxicity, and (5) high gas permeability. However, high hydrophobicity (water contact angle ~108°±7°) of PDMS often limits its applications where solutions containing biological samples are concerned. In our study, we created a simple, easy method for modifying the surface chemistry of PDMS microfluidic devices through the addition of surface-segregating additives during manufacture. In this method, a surface segregating copolymer is added to precursors for silicone and the desired device is manufactured following the usual methods. When the device surface is in contact with an aqueous solution, the copolymer self-organizes to expose its hydrophilic segments to the surface, making the surface of the silicone device more hydrophilic. This can lead to several improved performance criteria including lower fouling, lower non-specific adsorption, and better wettability. Specifically, this approach is expected to be useful for the manufacture of microfluidic devices. It is also likely to be useful for manufacturing silicone tubing and other materials, biomaterial applications, and surface coatings.

Keywords: microfluidics, non-specific protein adsorption, PDMS, PEG, copolymer

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Authors: Saleh Alshehri

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Thermoelectric technology is currently being used in many industrial applications for cooling, heating and generating electricity. This research mainly focuses on using thermoelectric to cool down high-speed computer chips at different operating conditions. A previously developed and validated three-dimensional model for optimizing and assessing the performance of cascaded thermoelectric and non-cascaded thermoelectric is used in this study to investigate the possibility of decreasing the hotspot temperature of computer chip. Additionally, a test assembly is built and tested at steady-state and transient conditions. The obtained optimum thermoelectric current at steady-state condition is used to conduct a number of pulsed tests (i.e. transient tests) with different shapes to cool the computer chips hotspots. The results of the steady-state tests showed that at hotspot heat rate of 15.58 W (5.97 W/cm²), using thermoelectric current of 4.5 A has resulted in decreasing the hotspot temperature at open circuit condition (89.3 °C) by 50.1 °C. Maximum and minimum hotspot temperatures have been affected by ON and OFF duration of the electrical current pulse. Maximum hotspot temperature was resulted by longer OFF pulse period. In addition, longer ON pulse period has generated the minimum hotspot temperature.

Keywords: thermoelectric generator, TEG, thermoelectric cooler, TEC, chip hotspots, electronic cooling

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Authors: Ruth Lorivi, Jianzheng Li, John J. Ambuchi, Kaiwen Deng

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Nitrogen removal from wastewater is accomplished by nitrification and denitrification processes. Successful denitrification requires carbon, therefore, if placed after biochemical oxygen demand (BOD) and nitrification process, a carbon source has to be re-introduced into the water. To avoid adding a carbon source, denitrification is usually placed before BOD and nitrification processes. This process however involves recycling the nitrified effluent. In this study wood chips were used as internal carbon source which enabled placement of denitrification after BOD and nitrification process without effluent recycling. To investigate the efficiency of a wood packed aerobic-anaerobic baffled reactor on carbon and nutrients removal from domestic wastewater, a three compartment baffled reactor was presented. Each of the three compartments was packed with 329 g wood chips 1x1cm acting as an internal carbon source for denitrification. The proposed mode of operation was aerobic-anoxic-anaerobic (OAA) with no effluent recycling. The operating temperature, hydraulic retention time (HRT), dissolved oxygen (DO) and pH were 24 ± 2 ℃, 24 h, less than 4 mg/L and 7 ± 1 respectively. The removal efficiencies of chemical oxygen demand (COD), ammonia nitrogen (NH₄⁺-N) and total nitrogen (TN) attained was 99, 87 and 83% respectively. TN removal rate was limited by nitrification as 97% of ammonia converted into nitrate and nitrite was denitrified. These results show that application of wood chips in wastewater treatment processes is an efficient internal carbon source. 

Keywords: aerobic-anaerobic baffled reactor, denitrification, nitrification, wood chip

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Authors: Andreea Campu, Ilinica Muresan, Simona Cainap, Simion Astilean, Monica Focsan

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Acute myocardial infarction (AMI) is the most severe cardiovascular disease, which has threatened human lives for decades, thus a continuous interest is directed towards the detection of cardiac biomarkers such as cardiac troponin I (cTnI) in order to predict risk and, implicitly, fulfill the early diagnosis requirements in AMI settings. Microfluidics is a major technology involved in the development of efficient sensing devices with real-time fast responses and on-site applicability. Microfluidic devices have gathered a lot of attention recently due to their advantageous features such as high sensitivity and specificity, miniaturization and portability, ease-of-use, low-cost, facile fabrication, and reduced sample manipulation. The integration of gold nanoparticles into the structure of microfluidic sensors has led to the development of highly effective detection systems, considering the unique properties of the metallic nanostructures, specifically the Localized Surface Plasmon Resonance (LSPR), which makes them highly sensitive to their microenvironment. In this scientific context, herein, we propose the implementation of a novel detection device, which successfully combines the efficiency of gold bipyramids (AuBPs) as signal transducers and thermal generators with the sample-driven advantages of the microfluidic channels into a miniaturized, portable, low-cost, specific, and sensitive test for the dual LSPR-thermographic cTnI detection. Specifically, AuBPs with longitudinal LSPR response at 830 nm were chemically synthesized using the seed-mediated growth approach and characterized in terms of optical and morphological properties. Further, the colloidal AuBPs were deposited onto pre-treated silanized glass substrates thus, a uniform nanoparticle coverage of the substrate was obtained and confirmed by extinction measurements showing a 43 nm blue-shift of the LSPR response as a consequence of the refractive index change. The as-obtained plasmonic substrate was then integrated into a microfluidic “Y”-shaped polydimethylsiloxane (PDMS) channel, fabricated using a Laser Cutter system. Both plasmonic and microfluidic elements were plasma treated in order to achieve a permanent bond. The as-developed microfluidic plasmonic chip was further coupled to an automated syringe pump system. The proposed biosensing protocol implicates the successive injection inside the microfluidic channel as follows: p-aminothiophenol and glutaraldehyde, to achieve a covalent bond between the metallic surface and cTnI antibody, anti-cTnI, as a recognition element, and target cTnI biomarker. The successful functionalization and capture of cTnI was monitored by LSPR detection thus, after each step, a red-shift of the optical response was recorded. Furthermore, as an innovative detection technique, thermal determinations were made after each injection by exposing the microfluidic plasmonic chip to 785 nm laser excitation, considering that the AuBPs exhibit high light-to-heat conversion performances. By the analysis of the thermographic images, thermal curves were obtained, showing a decrease in the thermal efficiency after the anti-cTnI-cTnI reaction was realized. Thus, we developed a microfluidic plasmonic chip able to operate as both LSPR and thermal sensor for the detection of the cardiac troponin I biomarker, leading thus to the progress of diagnostic devices.

Keywords: gold nanobipyramids, microfluidic device, localized surface plasmon resonance detection, thermographic detection

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Authors: Shashank Kumar, Mansi Chandra, Ujjawal Singh, Parth Gupta, Rishi Ram, Arnab Sarkar

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Milk is one of the basic and primary sources of food and energy as we start consuming milk from birth. Hence, milk quality and purity and checking the concentration of its constituents become necessary steps. Considering the importance of the purity of milk for human health, the following study has been carried out to simultaneously detect and quantify the different adulterants like urea and starch in milk with the help of a paper-based microfluidic device integrated with a smartphone. The detection of the concentration of urea and starch is based on the principle of colorimetry. In contrast, the fluid flow in the device is based on the capillary action of porous media. The microfluidic channel proposed in the study is equipped with a specialized detection zone, and it employs a colorimetric indicator undergoing a visible color change when the milk gets in touch or reacts with a set of reagents which confirms the presence of different adulterants in the milk. In our proposed work, we have used iodine to detect the percentage of starch in the milk, whereas, in the case of urea, we have used the p-DMAB. A direct correlation has been found between the color change intensity and the concentration of adulterants. A calibration curve was constructed to find color intensity and subsequent starch and urea concentration. The device has low-cost production and easy disposability, which make it highly suitable for widespread adoption, especially in resource-constrained settings. Moreover, a smartphone application has been developed to detect, capture, and analyze the change in color intensity due to the presence of adulterants in the milk. The low-cost nature of the smartphone-integrated paper-based sensor, coupled with its integration with smartphones, makes it an attractive solution for widespread use. They are affordable, simple to use, and do not require specialized training, making them ideal tools for regulatory bodies and concerned consumers.

Keywords: paper based microfluidic device, milk adulteration, urea detection, starch detection, smartphone application

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Authors: Rina Lee, Chung-Hun Oh, Eunjin Lee, Jeongyun Kim

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The Photodynamic therapy (PDT) is a treatment that uses a photosensitizer as a drug to damage and kill cancer cells. After injecting the photosensitizer into the bloodstream, the drug is absorbed by cancer cells selectively. Then the area to be treated is exposed to specific wavelengths of light and the photosensitizer produces a form of oxygen that kills nearby cancer cells. PDT is has an advantage to destroy the tumor with minimized side-effects on normal cells. But, PDT is not a completed method for cancer therapy. Because the mechanism of PDT is quite clear yet and the parameters such as intensity of light and dose of photosensitizer are not optimized for different types of cancers. To optimize these parameters, we suggest a novel microfluidic system to automatically control intensity of light exposure with a personal computer (PC). A polydimethylsiloxane (PDMS) microfluidic chip is composed with (1) a cell culture channels layer where cancer cells were trapped to be tested with various dosed photofrin (1μg/ml used for the test) as the photosensitizer and (2) a color dye layer as a neutral density (ND) filter to reduce intensity of light which exposes the cell culture channels filled with cancer cells. Eight different intensity of light (10%, 20%, …, 100%) are generated through various concentrations of blue dye filling the ND filter. As a light source, a light emitting diode (LED) with 635nm wavelength was placed above the developed PDMS microfluidic chip. The total time for light exposure was 30 minutes and HeLa and PC3 cell lines of cancer cells were tested. The cell viability of cells was evaluated with a Live/Dead assay kit (L-3224, Invitrogen, USA). The stronger intensity of light exposed, the lower viability of the cell was observed, and vice versa. Therefore, this system was demonstrated through investigating the PDT against cancer cell to optimize the parameters as critical light intensity and dose of photosensitizer. Our results suggest that the system can be used for optimizing the combinational parameters of light intensity and photosensitizer dose against diverse cancer cell types.

Keywords: photodynamic therapy, photofrin, high throughput screening, hela

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Authors: Martin Elstner

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Processing of information by means of handling chemicals is a ubiquitous phenomenon in nature. Technical implementations of chemical information processing lack of low integration densities compared to electronic devices. Stimuli responsive hydrogels are promising candidates for materials with information processing capabilities. These hydrogels are sensitive toward chemical stimuli like metal ions or amino acids. The binding of an analyte molecule induces conformational changes inside the polymer network and subsequently the water content and volume of the hydrogel varies. This volume change can control material flows, and concurrently information flows, in microfluidic devices. The combination of this technology with powerful chemical logic gates yields in a platform for highly integrated chemical circuits. The manufacturing process of such devices is very challenging and rapid prototyping is a key technology used in the study. 3D printing allows generating three-dimensional defined structures of high complexity in a single and fast process step. This thermoplastic master is molded into PDMS and the master is removed by dissolution in an organic solvent. A variety of hydrogel materials is prepared by dispenser printing of pre-polymer solutions. By a variation of functional groups or cross-linking units, the functionality of the hole circuit can be programmed. Finally, applications in the field of bio-molecular analytics were demonstrated with an autonomously operating microfluidic chip.

Keywords: bioanalytics, hydrogels, information processing, microvalve

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Authors: Itti Bist, Snehasis Bhakta, Di Jiang, Tia E. Keyes, Aaron Martin, Robert J. Forster, James F. Rusling

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DNA damage from metabolites of lipophilic drugs and pollutants, generated by enzymes, represents a major toxicity pathway in humans. These metabolites can react with DNA to form either 8-oxo-7,8-dihydro-2-deoxyguanosine (8-oxodG), which is the oxidative product of DNA or covalent DNA adducts, both of which are genotoxic and hence considered important biomarkers to detect cancer in humans. Therefore, detecting reactions of metabolites with DNA is an effective approach for the safety assessment of new chemicals and drugs. Here we describe a novel electrochemiluminescent (ECL) sensor array which can detect DNA oxidation and chemical DNA damage in a single array, facilitating a more accurate diagnostic tool for genotoxicity screening. Layer-by-layer assembly of DNA and enzyme are assembled on the pyrolytic graphite array which is housed in a microfluidic device for sequential detection of two type of the DNA damages. Multiple enzyme reactions are run on test compounds using the array, generating toxic metabolites in situ. These metabolites react with DNA in the films to cause DNA oxidation and chemical DNA damage which are detected by ECL generating osmium compound and ruthenium polymer, respectively. The method is further validated by the formation of 8-oxodG and DNA adduct using similar films of DNA/enzyme on magnetic bead biocolloid reactors, hydrolyzing the DNA, and analyzing by liquid chromatography-mass spectrometry (LC-MS). Hence, this combined DNA/enzyme array/LC-MS approach can efficiently explore metabolic genotoxic pathways for drugs and environmental chemicals.

Keywords: biosensor, electrochemiluminescence, DNA damage, microfluidic array

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Authors: Nizar Jawad Hadi, Sajad Abd Alabbas

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Because of their ability to encapsulate and release drugs in a controlled manner, microspheres fabricated from polymer emulsions using microfluidic devices have shown promise for drug delivery applications. In this study, the effects of velocity, density, viscosity, and surface tension, as well as channel diameter, on microsphere generation were investigated using Fluent Ansys software. The software was programmed with the physical properties of the polymer emulsion such as density, viscosity and surface tension. Simulation will then be performed to predict fluid flow and microsphere production and improve the design of drug delivery applications based on changes in these parameters. The effects of capillary and Weber numbers are also studied. The results of the study showed that the size of the microspheres can be controlled by adjusting the speed and diameter of the channel. Narrower microspheres resulted from narrower channel widths and higher flow rates, which could improve drug delivery efficiency, while smaller microspheres resulted from lower interfacial surface tension. The viscosity and density of the polymer emulsion significantly affected the size of the microspheres, ith higher viscosities and densities producing smaller microspheres. The loading and drug release properties of the microspheres created with the microfluidic technique were also predicted. The results showed that the microspheres can efficiently encapsulate drugs and release them in a controlled manner over a period of time. This is due to the high surface area to volume ratio of the microspheres, which allows for efficient drug diffusion. The ability to tune the manufacturing process using factors such as speed, density, viscosity, channel diameter, and surface tension offers a potential opportunity to design drug delivery systems with greater efficiency and fewer side effects.

Keywords: polymer emulsion, microspheres, numerical simulation, microfluidic device

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Authors: Andra Blumberga, Armands Gravelsins, Haralds Vigants, Dagnija Blumberga

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The paper discovers biotechonomy development analysis by use of system dynamics modelling. The research is connected with investigations of biomass application for production of bioproducts with higher added value. The most popular bioresource is wood, and therefore, the main question today is about future development and eco-design of products. The paper emphasizes and evaluates energy sector which is open for use of wood logs, wood chips, wood pellets and so on. The main aim for this research study was to build a framework to analyse development perspectives for wood pellet production. To reach the goal, a system dynamics model of energy wood supplies, processing, and consumption is built. Production capacity, energy consumption, changes in energy and technology efficiency, required labour source, prices of wood, energy and labour are taken into account. Validation and verification tests with available data and information have been carried out and indicate that the model constitutes the dynamic hypothesis. It is found that the more is invested into pellets production, the higher the specific profit per production unit compared to wood logs and wood chips. As a result, wood chips production is decreasing dramatically and is replaced by wood pellets. The limiting factor for pellet industry growth is availability of wood sources. This is governed by felling limit set by the government based on sustainable forestry principles.

Keywords: bioenergy, biotechonomy, system dynamics modelling, wood pellets

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Authors: Reza Hadjiaghaie Vafaie, Sevda Givtaj

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Automation and control of biological samples and solutions at the microscale is a major advantage for biochemistry analysis and biological diagnostics. Despite the known potential of miniaturization in biochemistry and biomedical applications, comparatively little is known about fluid automation and control at the microscale. Here, we study the electric field effect inside a fluidic channel and proper electrode structures with different patterns proposed to form forward, reversal, and rotational flows inside the channel. The simulation results confirmed that the ac electro-thermal flow is efficient for the control and automation of high-conductive solutions. In this research, the fluid pumping and mixing effects were numerically studied by solving physic-coupled electric, temperature, hydrodynamic, and concentration fields inside a microchannel. From an experimental point of view, the electrode structures are deposited on a silicon substrate and bonded to a PDMS microchannel to form a microfluidic chip. The motions of fluorescent particles in pumping and mixing modes were captured by using a CCD camera. By measuring the frequency response of the fluid and exciting the electrodes with the proper voltage, the fluid motions (including pumping and mixing effects) are observed inside the channel through the CCD camera. Based on the results, there is good agreement between the experimental and simulation studies.

Keywords: microfluidic, nano/micro actuator, AC electrothermal, Reynolds number, micropump, micromixer, microfabrication, mass transfer, biomedical applications

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Authors: Anil Koklu, Amin Mansoorifar, Ali Beskok

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Dielectric spectroscopy (DS) is a noninvasive, label free technique for a long term real-time measurements of the impedance spectra of biological cells. DS enables characterization of cellular dielectric properties such as membrane capacitance and cytoplasmic conductivity. We have developed a lab-on-a-chip device that uses an electro-activated microwells array for loading, DS measurements, and unloading of biological cells. We utilized from dielectrophoresis (DEP) to capture target cells inside the wells and release them after DS measurement. DEP is a label-free technique that exploits differences among dielectric properties of the particles. In detail, DEP is the motion of polarizable particles suspended in an ionic solution and subjected to a spatially non-uniform external electric field. To the best of our knowledge, this is the first microfluidic chip that combines DEP and DS to analyze biological cells using electro-activated wells. Device performance is tested using two different cell lines of prostate cancer cells (RV122, PC-3). Impedance measurements were conducted at 0.2 V in the 10 kHz to 40 MHz range with 6 s time resolution. An equivalent circuit model was developed to extract the cell membrane capacitance and cell cytoplasmic conductivity from the impedance spectra. We report the time course of the variations in dielectric properties of PC-3 and RV122 cells suspended in low conductivity medium (LCB), which enhances dielectrophoretic and impedance responses, and their response to sudden pH change from a pH of 7.3 to a pH of 5.8. It is shown that microfluidic chip allowed online measurements of dielectric properties of prostate cancer cells and the assessment of the cellular level variations under external stimuli such as different buffer conductivity and pH. Based on these data, we intend to deploy the current device for single cell measurements by fabricating separately addressable N × N electrode platforms. Such a device will allow time-dependent dielectric response measurements for individual cells with the ability of selectively releasing them using negative-DEP and pressure driven flow.

Keywords: microfluidic, microfabrication, lab on a chip, AC electrokinetics, dielectric spectroscopy

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Authors: Seul Ki Min, Gwang Heum Yoon, Jung Hyun Joo, Hwa Sung Shin

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Cellular membrane distortion is known as a factor to change intracellular signaling. However, progress of relevant studies is difficult because there are no facilities that can control membrane distortion finely. In this study, we developed microfluidic device which can inflict mechanical stress on cell membrane of Chlamydomonas reinhardtii using regular height of the channels. And cellular physiological changes were analyzed from cells cultured in the device. Excessive calcium ion influx through into cytoplasm was induced from mechanical stress. The results revealed that compressed cells had up-regulated Mat3 mRNA which regulates cell size and cell cycle from a prolonged G1 phase. Additionally, TAG used for the production of biodiesel was raised rapidly from 4 h after compression. Taken together, membrane distortion can be considered as an attractive inducer for biofuel production.

Keywords: mechanical stress, membrane distortion, Chlamydomonas reinhardtii, deflagellation, cell cycle, lipid metabolism

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Authors: Sourita Ghosh, Falguni Pati, Suhanya Duraiswamy

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Spheroid, a simple three-dimensional cellular aggregate, allows us to simulate the in-vivo complexity of cellular signaling and interactions in greater detail than traditional 2D cell culture. It can be used as an in-vitro model for drug toxicity testing, tumor modeling and many other such applications specifically for cancer. Our work is focused on the development of an affordable, user-friendly, robust, reproducible, high throughput microfluidic device for water in oil droplet production, which can, in turn, be used for spheroids manufacturing. Here, we have investigated the droplet breakup between two non-Newtonian fluids, viz. silicone oil and decellularized liver matrix, which acts as our extra cellular matrix (ECM) for spheroids formation. We performed some biochemical assays to characterize the liver ECM, as well as rheological studies on our two fluids and observed a critical dependence of capillary number (Ca) on droplet breakup and homogeneous drop formation

Keywords: chip less, droplets, extracellular matrix, liver spheroid

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Authors: C. Garcia, C. Andreasen

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Sugarcane is an important resource for bioenergy. Fields are usually established by using 15-20 cm pieces of sugarcane stalks as propagation material. An alternative method is to use small chips with nodes from sugarcane stalks. Plants from nodes are often established in plastic pots, but plastic pots could be replaced with biodegradable paper pots. This would be a more sustainable solution, reducing labor costs and avoiding pollution with plastic. We compared the establishment of plants from nodes taken from three different part of the sugarcane plant. The nodes were planted in plastic and paper pots. There was no significant difference between plants established in the two pot types. Nodes from different part of the stalk had different sprouting capacity. Nodes from the top parts sprouted significantly better than nodes taken from the middle or nodes taken closed to the ground in two experiments. Nodes with a length of 3 cm performed better than nodes with a length of 2 cm.

Keywords: nodes, paper pots, propagation material, sugarcane

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Authors: Patricia B. Cruz, Catrina Jean G. Valenzuela, Analyn N. Yumang

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Nearly a hundred per million of the Filipino population is diagnosed with Chronic Kidney Disease (CKD). The early stage of CKD has no symptoms and can only be discovered once the patient undergoes urinalysis. Over the years, different methods were discovered and used for the quantification of the urinary albumin such as the immunochemical assays where most of these methods require large machinery that has a high cost in maintenance and resources, and a dipstick test which is yet to be proven and is still debated as a reliable method in detecting early stages of microalbuminuria. This research study involves the use of the liquid chromatography concept in microfluidic instruments with biosensor as a means of separation and detection respectively, and linear regression to quantify human urinary albumin. The researchers’ main objective was to create a miniature system that quantifies and detect patients’ urinary albumin while reducing the amount of volume used per five test samples. For this study, 30 urine samples of unknown albumin concentrations were tested using VITROS Analyzer and the microfluidic system for comparison. Based on the data shared by both methods, the actual vs. predicted regression were able to create a positive linear relationship with an R² of 0.9995 and a linear equation of y = 1.09x + 0.07, indicating that the predicted values and actual values are approximately equal. Furthermore, the microfluidic instrument uses 75% less in total volume – sample and reagents combined, compared to the VITROS Analyzer per five test samples.

Keywords: Chronic Kidney Disease, Linear Regression, Microfluidics, Urinary Albumin

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Authors: E. Obreque-Slier, J. Laqui-Estaña, A. Peña-Neira, M. Medel-Marabolí

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Wine is an important source of polyphenols. Red wines show important concentrations of nonflavonoid (gallic acid, ellagic acid, caffeic acid and coumaric acid) and flavonoid compounds [(+)-catechin, (-)-epicatechin, (+)-gallocatechin and (-)-epigallocatechin]. However, a significant variability in the quantitative and qualitative distribution of chemical constituents in wine has to be expected depending on an array of important factors, such as the varietal differences of Vitis vinifera and cultural practices. It has observed that Carménère grapes present a differential composition and evolution of phenolic compounds when compared to other varieties and specifically with Cabernet Sauvignon grapes. Likewise, among the cultural practices, the aging in contact with oak wood is a high relevance factor. Then, the extraction of different polyphenolic compounds from oak wood into wine during its ageing process produces both qualitative and quantitative changes. Recently, many new techniques have been introduced in winemaking. One of these involves putting new pieces of wood (oak chips or inner staves) into inert containers. It offers some distinct and previously unavailable flavour advantages, as well as new options in wine handling. To our best knowledge, there is not information about the behaviour of Carménère wines (Chilean emblematic cultivar) in contact with oak wood. In addition, the effect of aging time and wood product (barrels, chips or staves) on the phenolic composition in Carménère wines has not been studied. This study aims at characterizing the condensed and hydrolyzable tannins from Carménère wines during the aging with staves, chips and barrels from French oak wood. The experimental design was completely randomized with two independent assays: aging time (0-12 month) and different formats of wood (barrel, chips and staves). The wines were characterized by spectrophotometric (total tannins and fractionation of proanthocyanidins into monomers, oligomers and polymers) and HPLC-DAD (ellagitannins) analysis. The wines in contact with different products of oak wood showed a similar content of total tannins during the study, while the control wine (without oak wood) presented a lower content of these compounds. In addition, it was observed that the polymeric proanthocyanidin fraction was the most abundant, while the monomeric fraction was the less abundant fraction in all treatments in two sample. However, significative differences in each fractions were observed between wines in contact from barrel, chips, and staves in two sample dates. Finally, the wine from barrels presented the highest content of the ellagitannins from the fourth to the last sample date. In conclusion, the use of alternative formats of oak wood affects the chemical composition of wines during aging, and these enological products are an interesting alternative to contribute with tannins to wine.

Keywords: enological inputs, oak wood aging, polyphenols, red wine

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Authors: Amina Farooq, Nauman Zafar Butt

Abstract:

This paper is about methods and tools for counting particles of interest, such as cells. A microfluidic system with interconnected electronics on a flexible substrate, inlet-outlet ports and interface schemes, sensitive and selective detection of cells specificity, and processing of cell counting at polymer interfaces in a microscale biosensor for use in the detection of target biological and non-biological cells. The development of fluidic channels, planar fluidic contact ports, integrated metal electrodes on a flexible substrate for impedance measurements, and a surface modification plasma treatment as an intermediate bonding layer are all part of the fabrication process. Magnetron DC sputtering is used to deposit a double metal layer (Ti/Pt) over the polypropylene film. Using a photoresist layer, specified and etched zones are established. Small fluid volumes, a reduced detection region, and electrical impedance measurements over a range of frequencies for cell counts improve detection sensitivity and specificity. The procedure involves continuous flow of fluid samples that contain particles of interest through the microfluidic channels, counting all types of particles in a portion of the sample using the electrical differential counter to generate a bipolar pulse for each passing cell—calculating the total number of particles of interest originally in the fluid sample by using MATLAB program and signal processing. It's indeed potential to develop a robust and economical kit for cell counting in whole-blood samples using these methods and similar devices.

Keywords: impedance, biochip, cell counting, microfluidics

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Authors: R. Vasireddi, J. Kruse, M. Vakili, M. Trebbin

Abstract:

Here we present a gas dynamic virtual nozzle (GDVN) based microfluidic jetting devices for spinning of nano/microfibers. The device is fabricated by soft lithography techniques and is based on the principle of a GDVN for precise three-dimensional gas focusing of the spinning solution. The nozzle device is used to produce micro/nanofibers of a perfluorinated terpolymer (THV), which were collected on an aluminum substrate for scanning electron microscopy (SEM) analysis. The influences of air pressure, polymer concentration, flow rate and nozzle geometry on the fiber properties were investigated. It was revealed that surface properties are controlled by air pressure and polymer concentration while the diameter and shape of the fibers are influenced mostly by the concentration of the polymer solution and pressure. Alterations of the nozzle geometry had a negligible effect on the fiber properties, however, the jetting stability was affected. Round and flat fibers with differing surface properties from craters, grooves to smooth surfaces could be fabricated by controlling the above-mentioned parameters. Furthermore, the formation of surface roughness was attributed to the fast evaporation rate and velocity (mis)match between the polymer solution jet and the surrounding air stream. The diameter of the fibers could be tuned from ~250 nm to ~15 µm. Because of the simplicity of the setup, the precise control of the fiber properties, access to biocompatible nanofiber fabrication and the easy scale-up of parallel channels for high throughput, this method offers significant benefits compared to existing solution-based fiber production methods.

Keywords: gas dynamic virtual nozzle (GDVN) principle, microfluidic device, spinning, uniform nanofibers

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Authors: Mitesh Rathod, Jungho Ahn, Noo Li Jeon, Junghoon Lee

Abstract:

Endothelial cells respond to cues from both biochemical as well as micro mechanical environment. Significant effort has been directed to understand the effects of biochemical signaling, however, relatively little is known about regulation of endothelial cell biology by the micro mechanical environment. Numerous studies have been performed to understand how physical forces regulate endothelial cell behavior. In this regard, past studies have majorly focused on exploring how fluid shear stress governs endothelial cell behavior. Parallel plate flow chambers and rectangular microchannels are routinely employed for applying fluid shear force on endothelial cells. However, these studies fall short in mimicking the in vivo like micro environment from topological aspects. Few studies have only used circular microchannels to replicate in vivo like condition. Seldom efforts have been directed to elucidate the combined effect of topology, substrate rigidity and fluid shear stress on endothelial cell response. In this regard, we demonstrate a facile fabrication process to develop a hybrid polydimethylsiloxane microfluidic platform to study endothelial cell biology. On a single chip microchannels with different cross sections i.e., circular, rectangular and square have been fabricated. In addition, our fabrication approach allows variation in the substrate rigidity along the channel length. Two different variants of polydimethylsiloxane, namely Sylgard 184 and Sylgard 527, were utilized to achieve the variation in rigidity. Moreover, our approach also enables in creating Y bifurcation circular microchannels. Our microfluidic platform thus facilitates for conducting studies pertaining to endothelial cell morphology with respect to change in topology, substrate rigidity and fluid flow on a single chip. The hybrid platform was tested by culturing Human Umbilical Vein Endothelial Cells in circular microchannels with varying substrate rigidity, and exposed to fluid shear stress of 12 dynes/cm² and static conditions. Results indicate the cell area response to flow induced shear stress was governed by the underlying substrate mechanics.

Keywords: hybrid, microfluidic platform, PDMS, shear flow, substrate rigidity

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Authors: V. Karamzadeh, J. Adhvaryu, A. Chandrasekaran, M. Packirisamy

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In Bulk Acoustic Wave (BAW) microfluidics, the throughput of particle sorting is dependent on the complex interplay between the geometric configuration of the channel, the size of the particles, and the properties of the fluid medium, which therefore calls for a detailed modeling and understanding of the fluid-particle interaction dynamics under an acoustic field, prior to designing the system. In this work, we propose a simplified Bulk acoustophoretic system that can be used for size dependent particle sorting. A Finite Element Method (FEM) based analytical model has been developed to study the dependence of particle sizes on channel parameters, and the sorting efficiency in a given fluid medium. Based on the results, the microfluidic system has been designed to take into account all the variables involved with the underlying physics, and has been fabricated using an additive manufacturing technique employing a commercial 3D printer, to generate a simple, cost-effective system that can be used for size sensitive particle sorting.

Keywords: 3D printing, 3D microfluidic chip, acoustophoresis, cell separation, MEMS (Microelectromechanical Systems), microfluidics

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Authors: Soroosh Torabi, Brad Berron, Ren Xu, Christine Trinkle

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Microfluidics integrated with 3D cell culture is a powerful technology to mimic cellular environment, and can be used to study cell activities such as proliferation, migration and response to drugs. This technology has gained more attention in cancer studies over the past years, and many organ-on-a-chip systems have been developed to study cancer cell behaviors in an ex-vivo tumor microenvironment. However, there are still some barriers to adoption which include low throughput, complexity in 3D cell culture integration and limitations on non-optical analysis of cells. In this study, a user-friendly microfluidic multi-well plate was developed to mimic the in vivo tumor microenvironment. The microfluidic platform feeds multiple 3D cell culture sites at the same time which enhances the throughput of the system. The platform uses hydrophobic Cassie-Baxter surfaces created by microchannels to enable convenient loading of hydrogel/cell suspensions into the device, while providing barrier free placement of the hydrogel and cells adjacent to the fluidic path. The microchannels support convective flow and diffusion of nutrients to the cells and a removable lid is used to enable further chemical and physiological analysis on the cells. Different breast cancer cell lines were cultured in the device and then monitored to characterize nutrient delivery to the cells as well as cell invasion and proliferation. In addition, the drug response of breast cancer cell lines cultured in the device was compared to the response in xenograft models to the same drugs to analyze relevance of this platform for use in future drug-response studies.

Keywords: microfluidics, multi-well 3d cell culture, tumor microenvironment, tumor-on-a-chip

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Authors: Muhammad Rosli Abdullah, Noor Hasmiza Harun

Abstract:

A microorganism detection system has a potential to be used with the advancement in a biosensor development. The detection system requires an optical sensing system, microfluidic device and biological reagent. Although, the biosensors are available in the market, a label free and a lab-on-chip approach will promote a flexible solution. As a preliminary study of microorganism detection, three mechanisms such as Total Internal Reflection (TIR), Micro Fluidic Channel (MFC) and magnetic-electric field propagation were study and simulated. The objective are to identify the TIR angle, MFC parabolic flow and the wavelength for the microorganism detection. The simulation result indicates that evanescent wave is achieved when TIR angle > 42°, the corner and centre of a parabolic velocity are 0.02 m/s and 0.06 m/s respectively, and a higher energy distribution of a perfect electromagnetic scattering with dipole resonance radiation occurs at 500 nm. This simulation is beneficial to determine the components of the microorganism detection system that does not rely on classical microbiological, immunological and genetic methods which are laborious, time-consuming procedures and confined to specialized laboratories with expensive instrumentation equipment.

Keywords: microorganism, microfluidic, total internal reflection, lab on chip

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Authors: Yusser Olguín, Diego Benavente, Fernando Dorta, Nicole Orellana, Cristian Acevedo

Abstract:

One of the greatest challenges of tissue engineering is the generation of materials in which the highest possible number of conditions can be incorporated to stimulate the proliferation and differentiation of cells, which will be transformed together with the material into new functional tissue. In this sense, considering the properties of microfluidics and its relationship with cellular micro-environments, the possibility of controlling flow patterns and the ability to design diverse patterns in the chips, a microfluidic cell culture system can be established as a means for the evaluation of the effect of different parameters in a controlled and precise manner. Specifically in relation to the study and development of alternatives in peripheral nervous tissue engineering, it is necessary to consider different physical and chemical neurotrophic stimuli that promote cell growth and differentiation. Chemical stimuli include certain vitamins, glucocorticoids, gangliosides, and growth factors, while physical stimuli include topological stimuli, mechanical forces of the cellular environment and electrical stimulation. In this context, the present investigation shows the results of cell stimulation in a microbioreactor using electrical and chemical stimuli, where the differentiation of PC12 cells as a neuronal model is evidenced by neurite expression, dependent on the stimuli and their combination. The results were analysed with a multi-factor statistical approach, showing several relationships and dependencies between different parameters. Chip design, operating parameters and concentrations of neurotrophic chemical factors were found to be preponderant, based on the characteristics of the electrical stimuli.

Keywords: microfluidics, nerve tissue engineering, microbioreactor, electrical stimuli

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Authors: M. Mashiur Rahman, Ulrik Birk Henriksen, Jesper Ahrenfeldt, Maria Puig Arnavat

Abstract:

Using biomass gasification to make producer gas is one of the promising sustainable energy options available for small scale plant and rural applications for power and electricity. Tar content in producer gas is the main problem if it is used directly as a fuel. A low-tar biomass (LTB) gasifier of approximately 30 kW capacity has been developed to solve this. Moving bed gasifier with internal recirculation of pyrolysis gas has been the basic principle of the LTB gasifier. The gasifier focuses on the concept of mixing the pyrolysis gases with gasifying air and burning the mixture in separate combustion chamber. Five tests were carried out with the use of wood pellets and wood chips separately, with moisture content of 9-34%. The LTB gasifier offers excellent opportunities for handling extremely low-tar in the producer gas. The gasifiers producer gas had an extremely low tar content of 21.2 mg/Nm³ (avg.) and an average lower heating value (LHV) of 4.69 MJ/Nm³. Tar content found in different tests in the ranges of 10.6-29.8 mg/Nm³. This low tar content makes the producer gas suitable for direct use in internal combustion engine. Using mass and energy balances, the average gasifier capacity and cold gas efficiency (CGE) observed 23.1 kW and 82.7% for wood chips, and 33.1 kW and 60.5% for wood pellets, respectively. Average heat loss in term of higher heating value (HHV) observed 3.2% of thermal input for wood chips and 1% for wood pellets, where heat loss was found 1% of thermal input in term of enthalpy. Thus, the LTB gasifier performs better compared to typical gasifiers in term of heat loss. Equivalence ratio (ER) in the range of 0.29 to 0.41 gives better performance in terms of heating value and CGE. The specific gas production yields at the above ER range were in the range of 2.1-3.2 Nm³/kg. Heating value and CGE changes proportionally with the producer gas yield. The average gas compositions (H₂-19%, CO-19%, CO₂-10%, CH₄-0.7% and N₂-51%) obtained for wood chips are higher than the typical producer gas composition. Again, the temperature profile of the LTB gasifier observed relatively low temperature compared to typical moving bed gasifier. The average partial oxidation zone temperature of 970°C observed for wood chips. The use of separate combustor in the partial oxidation zone substantially lowers the bed temperature to 750°C. During the test, the engine was started and operated completely with the producer gas. The engine operated well on the produced gas, and no deposits were observed in the engine afterwards. Part of the producer gas flow was used for engine operation, and corresponding electrical power was found to be 1.5 kW continuously, and maximum power of 2.5 kW was also observed, while maximum generator capacity is 3 kW. A thermodynamic equilibrium model is good agreement with the experimental results and correctly predicts the equilibrium bed temperature, gas composition, LHV of the producer gas and ER with the experimental data, when the heat loss of 4% of the energy input is considered.

Keywords: biomass gasification, low-tar biomass gasifier, tar elimination, engine, deposits, condensate

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