This research project was part of the microfluidic program in the CRC for Microtechnology, Australia, during 2000 to 2003. The aim of this research was to investigate the feasibility of applying geometric variations in a microchannel to create effects other than pure molecular diffusion to enhance microfluidic mixing. Geometric variations included the shape of a microchannel, as well as the various obstacle structures inside the microchannel. Generally, before performing chemical or biological analysis, samples and reagents need to be mixed together thoroughly. This is particularly important in miniaturized Total Analysis Systems (�TAS), where mixing is critical for the detection stage. In scaling down dimensions of micro-devices, diffusion becomes an efficient method for achieving homogenous solutions when the characteristic length of the channels becomes sufficiently small. In the case of pressure driven flow, it is necessary to use wider microchannels to ensure fluids can be pumped through the channels and the volume of fluid can provide sufficient signal intensity for detection. However, a relatively wide microchannel makes mixing by virtue of pure molecular diffusion a very slow process in a confined volume of a microfluidic device. Therefore, mixing is a challenge and improved methods need to be found for microfluidic applications. In this research, passive mixing using geometric variations in microchannels was studied due to its advantages over active mixing in terms of simplicity and ease of fabrication. Because of the nature of laminar flow in a microchannel, the geometric variations were designed to improve lateral convection to increase cross-stream diffusion. Previous research using this approach was limited, and a detailed research program using computational fluid dynamic (CFD) solvers, various shapes, sizes and layouts of geometric structures was undertaken for the first time. Experimental measurements, published experimental data and analytical predictions were used to validate the simulations for selected samples. Mixing efficiency was evaluated by using mass fraction distributions. It was found that the overall performance of a micromixer should include the pressure drop in a microdevice, therefore, a mixing index criterion was formulated in this research to combine the effect of mixing efficiency and pressure drop. The mixing index was used to determine optimum parameters for enhanced mixing, as well as establish design guidelines for such devices. Three types of geometric variations were researched. First, partitioning in channels was used to divide fluids into mixing zones with different concentrations. Various designs were investigated, and while these provided many potential solutions to achieving good mixing, they were difficult to fabricate. Secondly, structures were used to create lateral convection, or secondary flows. Most of the work in this category used obstacles to disrupt the flow. It was found that symmetric layouts of obstacles in a channel had little effect on mixing, whereas, asymmetric arrangements created lateral convection to enhance crossstream diffusion and increase mixing. Finally, structures that could create complex 3D advections were investigated. At high Reynolds numbers (Re = 50), 3D ramping or obstacles generated strong lateral convection. Microchannels with 3D slanted grooves were also investigated. Mixers with grooved surfaces generated helicity at low Reynolds numbers (Re � 5) and provided a promising way to reduce the diffusion path in microchannels by stretching and folding of fluid streams. Deeper grooves resulted in better mixing efficiency. The 3D helical advection created by the patterned grooves in a microchannel was studied by using particle tracing algorithms developed in this research to generate streaklines and Poincare maps, which were used to evaluate the mixing performance. The results illustrated that all the types of mixers could provide solutions to microfluidic mixing when dimensional parameters were optimized.

No description available.

Conventional cell cytometers are often bulky and thus not convenient for bio-medical analysis where portable devices are desired. They also suffer from the drawback of high cost due to the complicated and expensive optical detection system involved. Therefore miniaturizing conventional cell cytometer is highly demanded as it offers an opportunity to transform the conventional bulky systems to more cost-efficient and portable microfluidic cell sorting devices. In addition to the advantages reduced cost and enhanced portability, microfluidic cell sorting devices require only a tiny amount of sample for analysis. In this thesis, one common microfluidic cell sorting device is developed using similar conventional functions and concepts but different sorting method. Unlike most of the conventional cell cytometers in which an electrical field or magnetic field is employed to deflect the charged target cells to the collecting container, microfluidic cell sorting devices use the fluid flow to control the movement of the targeted cells to the collecting reservoir. By using an electroosmitic pump, the response time of the flow switch is significantly lowered, leading to a much higher sorting efficiency. Despite the advantages of microfluidic cell sorting devices, there are some issues need to be addressed before realization of such devices. For example, more studies are required on the successful integration of the optical elements in the devices. In microfluidic, the transport phenomena is also different from that in macroscopic. Unlike that in macroscopic, surface forces are important in microfluidics. They result in pressure-induced flow which gives the parabolic profile of the velocity along the channel. Also, a plug-like velocity which is generated by the electoosmitic flow is required for the more controllable and accurate detection. To suppress the pressure-driven flow, hydro-resistance elements (Shallow channel network) are implemented on the microfluidic devices. Fabrication of optical elements by deposition of optical materials on glass or silicon wafer has been reported. However, this Micro Electro-Mechanical (MEM) technique requires special equipment and cleanroom facilities used in the semiconductor industry. A good alternative to the MEMS technique is soft lithography where optical elements can be created using polymers. In this work, ultraviolet-sensitive photo resists SU8 is used to fabricate the microfluidic cell sorting devices and the optical elements. By using the mask with the patterns of the microchannel network and optical elements, the optical elements can be fabricated with the microchannel, eliminating the problem of alignment. Experiments are also conducted to evaluate the integrated optical elements. To prevent cross-contamination, samples are usually prepared and are only mixed inside the microfluidic devices by the embedded mixers. Such embedded mixers, however, pose a great challenge as the small characteristic length of a microfluidic device tends to give a laminar flow and diffusion-dominated mixing. A simple passive micromixer is investigated to find the possibilities to integrate it to the microfluidic devices. To truly understand the diffusional mixing, a Y channel mixer is studied through the numerical and experimental investigations. Based on the results found, a possible design is also proposed and evaluated by experiments.

Conventional cell cytometers are often bulky and thus not convenient for bio-medical analysis where portable devices are desired. They also suffer from the drawback of high cost due to the complicated and expensive optical detection system involved. Therefore miniaturizing conventional cell cytometer is highly demanded as it offers an opportunity to transform the conventional bulky systems to more cost-efficient and portable microfluidic cell sorting devices. In addition to the advantages reduced cost and enhanced portability, microfluidic cell sorting devices require only a tiny amount of sample for analysis. In this thesis, one common microfluidic cell sorting device is developed using similar conventional functions and concepts but different sorting method. Unlike most of the conventional cell cytometers in which an electrical field or magnetic field is employed to deflect the charged target cells to the collecting container, microfluidic cell sorting devices use the fluid flow to control the movement of the targeted cells to the collecting reservoir. By using an electroosmitic pump, the response time of the flow switch is significantly lowered, leading to a much higher sorting efficiency. Despite the advantages of microfluidic cell sorting devices, there are some issues need to be addressed before realization of such devices. For example, more studies are required on the successful integration of the optical elements in the devices. In microfluidic, the transport phenomena is also different from that in macroscopic. Unlike that in macroscopic, surface forces are important in microfluidics. They result in pressure-induced flow which gives the parabolic profile of the velocity along the channel. Also, a plug-like velocity which is generated by the electoosmitic flow is required for the more controllable and accurate detection. To suppress the pressure-driven flow, hydro-resistance elements (Shallow channel network) are implemented on the microfluidic devices. Fabrication of optical elements by deposition of optical materials on glass or silicon wafer has been reported. However, this Micro Electro-Mechanical (MEM) technique requires special equipment and cleanroom facilities used in the semiconductor industry. A good alternative to the MEMS technique is soft lithography where optical elements can be created using polymers. In this work, ultraviolet-sensitive photo resists SU8 is used to fabricate the microfluidic cell sorting devices and the optical elements. By using the mask with the patterns of the microchannel network and optical elements, the optical elements can be fabricated with the microchannel, eliminating the problem of alignment. Experiments are also conducted to evaluate the integrated optical elements. To prevent cross-contamination, samples are usually prepared and are only mixed inside the microfluidic devices by the embedded mixers. Such embedded mixers, however, pose a great challenge as the small characteristic length of a microfluidic device tends to give a laminar flow and diffusion-dominated mixing. A simple passive micromixer is investigated to find the possibilities to integrate it to the microfluidic devices. To truly understand the diffusional mixing, a Y channel mixer is studied through the numerical and experimental investigations. Based on the results found, a possible design is also proposed and evaluated by experiments.

Experiments were conducted in electroosmotic flow (EOF) with 0.005≤Re ≤ 0.039 on mixing enhancement in 3-D helical microchannels. Both inlet velocity and concentration distribution along the flow channel were measurement via £gPIV and £gLIF technique respectively. The experimental results showed that the helical channels can generate nearly fully chaotic flow and achieve the complete mixing in a relatively short channel with three different helical channels (3, 4, and 6 inlet channels), and the four-inlet channel found to have the best mixing efficiency. Finally, the mixing length was correlated into a form of £f/Dh = 2.8Pe0.35 within ¡Ó8% accuracy between the experiments and prediction.

A microfluídica juntamente com a intensificação de processos são duas áreas de pesquisa interessadas no estudo e desenvolvimento de processos em escala micrométrica capazes de manipular diminutas quantidades de reagentes. Para tanto, estes devem contar com dispositivos de pequena escala de tamanho e ao mesmo tempo serem tão confiáveis e eficientes quanto os de escala industrial. Uma das operações unitárias envolvidas nesses processos é a agitação. Em função da ordem de grandeza dos equipamentos e dos materiais em que são fabricados, grandes diferenciais de pressão não podem ser aplicados nos mesmos e como conseqüência no interior dos micromisturadores, como são conhecidos tais equipamentos, o escoamento se dá em regime laminar, sob está condição o processo de mistura é controlado pela difusão entre os componentes. Uma maneira de superar esta dificuldade é gerar no interior do micromisturador o aparecimento de um escoamento caótico. Para tal, podem-se utilizar fontes de energia externa (micromisturadores ativos) ou a própria energia do escoamento (micromisturadores passivos) através da construção de geometrias especiais. O desenvolvimento em laboratório destes equipamentos demanda tempo e geralmente é oneroso. A principal alternativa para este trabalho é a dinâmica dos fluidos computacional (CFD), ferramenta aplicada no presente estudo para analisar três geometrias diferentes propostas e analisadas experimentalmente no trabalho de Cunha (2007). Para caracterizar o funcionamento dos mesmos foram testadas quatro vazões distintas, com as quais foi possível levantar os perfis de pressão, velocidade e fração mássica de dois componentes que eram misturados. Com o intuito de demonstrar a eficiência dos equipamentos dois parâmetros foram analisados: o avanço da qualidade da mistura e a perda de carga para as diferentes condições operacionais. Apesar da limitação da malha e de não ter-se obtido resultados independentes da malha, foi possível se fazer uma comparação entre as três geometrias e identificouse que os micromisturadores M2 e M3 são os que apresentam o melhor desempenho para a faixa de vazão simulada (120 < Re < 1200). / Microfluidics and process intensification are two research areas interested in the study and development of new micrometric-scale devices capable of manipulating and processing small quantities of reagents. These processes have to deal with small scale equipment and at the same time be as reliable and efficient as the large-scale one. Because of the scale of this equipment and the material it is made of, large pressure differential is not possible, as a consequence in the interior of the micromixers, as they are known; a laminar flow develops, under those circumstances the mixing process is controlled by the diffusion mechanism between the two components. One way to suppress this deficiency is to generate a chaotic flow on the micromixer, which can be done by using external energy (active micromixer) or its own flow energy (passive micromixer) through special geometry construction. The experimental development of such microdevices demands time and, generally, is very expensive. The main alternative for this activity is the use of computational fluid dynamics; this tool was employed on this work with the aim of studying three geometries proposed by Cunha (2007). To characterize their working process, four different volumetric flows were simulated and analyzed the pressure, velocity and mass fraction profiles. Two parameters were calculated in order to characterize their efficiency: the mixture quality along the micromixers cross sections and the pressure drop for different operational conditions. Although we have mesh size limitations and a mesh independent results were not obtained it was possible to compare the three micromixers geometries and it was found out that both M2 and M3 micromixers had the best performance under operational conditions tested (120 < Re < 1200).

A microfluídica juntamente com a intensificação de processos são duas áreas de pesquisa interessadas no estudo e desenvolvimento de processos em escala micrométrica capazes de manipular diminutas quantidades de reagentes. Para tanto, estes devem contar com dispositivos de pequena escala de tamanho e ao mesmo tempo serem tão confiáveis e eficientes quanto os de escala industrial. Uma das operações unitárias envolvidas nesses processos é a agitação. Em função da ordem de grandeza dos equipamentos e dos materiais em que são fabricados, grandes diferenciais de pressão não podem ser aplicados nos mesmos e como conseqüência no interior dos micromisturadores, como são conhecidos tais equipamentos, o escoamento se dá em regime laminar, sob está condição o processo de mistura é controlado pela difusão entre os componentes. Uma maneira de superar esta dificuldade é gerar no interior do micromisturador o aparecimento de um escoamento caótico. Para tal, podem-se utilizar fontes de energia externa (micromisturadores ativos) ou a própria energia do escoamento (micromisturadores passivos) através da construção de geometrias especiais. O desenvolvimento em laboratório destes equipamentos demanda tempo e geralmente é oneroso. A principal alternativa para este trabalho é a dinâmica dos fluidos computacional (CFD), ferramenta aplicada no presente estudo para analisar três geometrias diferentes propostas e analisadas experimentalmente no trabalho de Cunha (2007). Para caracterizar o funcionamento dos mesmos foram testadas quatro vazões distintas, com as quais foi possível levantar os perfis de pressão, velocidade e fração mássica de dois componentes que eram misturados. Com o intuito de demonstrar a eficiência dos equipamentos dois parâmetros foram analisados: o avanço da qualidade da mistura e a perda de carga para as diferentes condições operacionais. Apesar da limitação da malha e de não ter-se obtido resultados independentes da malha, foi possível se fazer uma comparação entre as três geometrias e identificouse que os micromisturadores M2 e M3 são os que apresentam o melhor desempenho para a faixa de vazão simulada (120 < Re < 1200). / Microfluidics and process intensification are two research areas interested in the study and development of new micrometric-scale devices capable of manipulating and processing small quantities of reagents. These processes have to deal with small scale equipment and at the same time be as reliable and efficient as the large-scale one. Because of the scale of this equipment and the material it is made of, large pressure differential is not possible, as a consequence in the interior of the micromixers, as they are known; a laminar flow develops, under those circumstances the mixing process is controlled by the diffusion mechanism between the two components. One way to suppress this deficiency is to generate a chaotic flow on the micromixer, which can be done by using external energy (active micromixer) or its own flow energy (passive micromixer) through special geometry construction. The experimental development of such microdevices demands time and, generally, is very expensive. The main alternative for this activity is the use of computational fluid dynamics; this tool was employed on this work with the aim of studying three geometries proposed by Cunha (2007). To characterize their working process, four different volumetric flows were simulated and analyzed the pressure, velocity and mass fraction profiles. Two parameters were calculated in order to characterize their efficiency: the mixture quality along the micromixers cross sections and the pressure drop for different operational conditions. Although we have mesh size limitations and a mesh independent results were not obtained it was possible to compare the three micromixers geometries and it was found out that both M2 and M3 micromixers had the best performance under operational conditions tested (120 < Re < 1200).

Integrated microfluidic devices for biomedical analysis attract lots of interest in the MEMS (Micro-Electro-Mechanical-Systems) research field. However, the characteristic Reynolds number for liquids flowing in these microchannels is very small (typically less than 10). At such low Reynolds numbers, turbulent mixing does not occur and homogenization of the solutions occurs through diffusion processes alone. Hence, a satisfactory mixing performance generally requires the use of extended flow channels and takes longer to accomplish such that the practical benefits of such devices are somewhat limited. Consequently, accomplishing the goal of u¡VTAS requires the development of enhanced mixing techniques for microfluidic structures. This study first presents a microfluidic mixer utilizing alternatively switching electroosmotic flow and proposes two microchannel designs of T-form and double-T-form micromixer. Switching DC field is used to generate the electroosmotic force to drive the fluid and also used for mixing of the fluids simultaneously, such that moving parts in the microfluidic device and delicate external control system are not required for the mixing purpose. Furthermore, this study also proposed a novel pinched-switching mode in the T-form microfluidic mixer, which could be effectively increase the perturbation within the fluid to promote the mixing efficiency. In this study, computer simulation for the operation conditions is used to predict the mixing outcomes and the mixing performance is also confirmed experimentally. Result shows the mixing performance can be as larger as 95% within the mixing distance of 1 mm downstream the common boundary between the different sample fluids. The novel method proposed in this study can be used for solving the mixing problem in a simple way in the field of micro-total-analysis-systems. Furthermore, in order to demonstrate the proposed micromixer is feasible for on-line bio-reaction, this study designs a fully integrated device for demonstration of DNA/enzyme reaction within the microfluidic chip. The microchip device contains a pre-column concentrating region, a micro mixer for DNA-enzyme mixing, an adjustable temperature control system and a post-column concentration channel. The integrated microfluidic chip has been used to implement the DNA digestion and extraction. Successfully digestion of £f-DNA using EcoRI restriction enzyme in the proposed device is demonstrated utilizing large-scale gel electrophoresis scheme. Results show that the reaction speed doubled while using the microfluidic system. In addition, on-line DNA digestion and capillary electrophoresis detection is also successfully demonstrated using a standard DNA-enzyme system of $X-174 and Hae III. Finally, this reasearch also proposes a novel cell/microparticle manipulation platform by integrating an optical tweezer system and a micro flow cytometer. During operation, electrokinetically driven sheath flows are utilized to focus microparticles to flow in the center of the sample stream then pass through an optical manipulation area. An IR diode laser is focused to generate force gradient in the optical manipulation area to manipulate the microparticles in the microfluidic device. Moving the particles at a static condition is demonstrated to confirm the feasibility of the home-built optical tweezer. The trapping force of the optical tweezer is measured using a novel method of Stocks-drag equilibrium. The proposed system can continuously catch moving microparticles in the flowing stream or switch them to flow into another sample flow within the microchannel. Target particles can be separated from the sample particles with this high efficient approach. More importantly, the system demonstrates a continuously manipulation of microparticles using non-contact force gradient such that moving parts and delicate fabrication processes can be excluded. The proposed system is feasible of high-throughput catching, moving, manipulation and sorting specific microparticles/cells within a mixed sample and results in a simple solution for cell/microparticle manipulation in the field of micro-total-analysis-systems. In this thesis, low-cost soda-lime glass substrates are adopted for the microchip fabrication using a simple and reliable fabrication process. Three kinds of novel microfluidic devices including an electrokinetically-driven microfluidic mixer, a high throughput DNA/enzyme reactor and an optically cell manipulation platform are successfully demonstrated. It is the author¡¦s believes that the results of this study will give important contributions in the development of micro-total-analysis-systems in the future. With the success of this study, we have a further step approaching to the dream of lab-on-a-chip system for bio-analytical applications.

This study proposed a Y-type mixer which was driven by electroosmotic flow (Ex = 5 - 25 kV/m) with 7 different mixing angles (30¢X, 60¢X, 90¢X, 120¢X, -120¢X, -90¢X, -60¢X) to enhance mixing efficiency . The mixing performance of the device was demonstrated by using micro laser-induced fluorescence (£gLIF) technology to quantify the concentration distribution in the microchannel. Also, micro particle image velocimetry (£gPIV) was used for velocity measurements and analysis. It was found that the negative mixing angle could induce larger dead zone area than the positive one. The joule heating effect was found when electric field strength was larger than 15 kV/m. The combined dead zone and joule heating effect could enhance the mixing performance slightly. Although it has only a marginal effect on the mixing length for the positive mixing angles. Negative mixing angles allow a reduction of mixer size, which means a more efficient use of material and space. Finally, the best mixing angle was found to be -60¢X.

京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第21075号 / 工博第4439号 / 新制||工||1690(附属図書館) / 京都大学大学院工学研究科化学工学専攻 / (主査)教授 前 一廣, 教授 吉田 潤一, 教授 長谷部 伸治 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DFAM