Theoretical and numerical studies of chaotic mixing are performed to circumvent the difficulties of efficient mixing, which come from the lack of turbulence in microfluidic devices. In order to carry out efficient and accurate parametric studies and to identify a fully chaotic state, a spectral element algorithm for solution of the incompressible Navier-Stokes and species transport equations is developed. Using Taylor series expansions in time marching, the new algorithm employs an algebraic factorization scheme on multi-dimensional staggered spectral element grids, and extends classical conforming Galerkin formulations to nonconforming spectral elements. Lagrangian particle tracking methods are utilized to study particle dispersion in the mixing device using spectral element and fourth order Runge-Kutta discretizations in space and time, respectively. Comparative studies of five different techniques commonly employed to identify the chaotic strength and mixing efficiency in microfluidic systems are presented to demonstrate the competitive advantages and shortcomings of each method. These are the stirring index based on the box counting method, Poincare sections, finite time Lyapunov exponents, the probability density function of the stretching field, and mixing index inverse, based on the standard deviation of scalar species distribution. Series of numerical simulations are performed by varying the Peclet number (Pe) at fixed kinematic conditions. The mixing length (lm) is characterized as function of the Pe number, and lm ∝ ln(Pe) scaling is demonstrated for fully chaotic cases. Employing the aforementioned techniques, optimum kinematic conditions and the actuation frequency of the stirrer that result in the highest mixing/stirring efficiency are identified in a zeta potential patterned straight micro channel, where a continuous flow is generated by superposition of a steady pressure driven flow and time periodic electroosmotic flow induced by a stream-wise AC electric field. Finally, it is shown that the invariant manifold of hyperbolic periodic point determines the geometry of fast mixing zones in oscillatory flows in two-dimensional cavity.

Lipid bi-layers are ubiquitous components of biological cells and are found in a variety of cell components. In biological membranes, lipid bi-layer membranes carry membrane proteins, which control transport of material and communication of signals in and out the cell. There are several disadvantages involved with patch clamping method as a way of studying biological membranes and protein interactions. Hence, artificial synthesis of bi-layer has been of great interest in basic biophysical studies, drug discoveries in pharmaceutical studies and study of protein nanopores for precise engineering applications. However, conventional lipid bi-layer synthesis techniques require skilled operators, have low repeatability (reliability), have portability restrictions and result in unstable bi-layers having a short lifetime. In this investigation a novel microfluidic device and a method for artificial synthesis of lipid bi-layer in-situ are explored. In the proposed method, lipid trapped at an aperture on a Teflon sheet, is thinned to form a lipid bi-layer by a continuous flow of buffer solution on both sides of the aperture in the microfluidic device. The microfluidic device is expected to have advantages from its compact design. Further, the new approach is expected to be repetitive and good for automation removing the requirement of a skilled operator. The microfluidic device was fabricated using two glass substrates. Two channels of ? ? shape were etched and through holes were fabricated at all four terminal ends of the microchannels on each glass substrate. A thin Teflon sheet carrying a 100?m diameter hole was sandwiched between the two glass wafers forming two sets of microchannels on both sides of the aperture. An analytical microfluid model of the microchannels was developed to investigate the nature of the flow and to select microchannel parameters. Experiments using the proposed device were performed to verify the feasibility of the novel approach for lipid bi-layer synthesis. Experimental results suggest formation of a lipid bi-layer at an aperture on the Teflon sheet but further investigation might be necessary for verification. Life time of the bi-layer is short mainly due to low quality of the used aperture.

Thesis (M.S.)--Oregon State University, 2010. / Printout. Includes bibliographical references (leaves 77-78). Also available on the World Wide Web.

Controlled drug delivery devices provide numerous advantages such as reduced side effects, higher therapeutic efficiency and improved patient compliance. Biodegradable polymer has become the most important material for controlled drug delivery device because of the excellent biocompatibility and tunable physicochemical properties. Biodegradable polymeric drug delivery devices are usually processed into various types of micro-particles due to the ease of fabrication and administration. However, controlling the drug release kinetics of these microparticles is still a challenge. One important reason is that drug release kinetics is significantly influenced by the microstructure of drug delivery devices, which is difficult to control. 　Microfluidic method is a group of technologies involved in the manipulation of fluids using channels in the scale of micrometers. Microfluidic method is particularly useful in controlling the structure of micro-droplets and generating homogeneous droplets. Therefore, microfluidics suggests great potential in controlling microstructures of drug delivery devices and drug release kinetics. 　In this study, biodegradable polymer based controlled drug delivery devices were fabricated using microfluidic method. Various types of microstructures were developed such as microspheres, core-shell microspheres, hollow microspheres and hydrogel microspheres. The results showed that microstructures were well controlled by fluid flow rates and geometries of capillary microfluidic devices. Both hydrophobic and hydrophilic drugs could be delivered by choosing drug delivery devices with suitable microstructures. 　Drug release kinetics of biodegradable polymeric microspheres has been studies a lot, yet complete understanding is still to be achieved. The diameter is an important factor which contributes to the drug release kinetics. However, the influence of diameter has not been systemically studied because monodisperse microspheres are difficult to obtain. Using microfluidic method, monodisperse PLGA microspheres with different diameters were fabricated to study the influence of diameter on drug release kinetics. It was found that diameter only influence the duration of the first phase (lag phase) in drug release process and smaller microspheres exhibited shorter lag phase. The relatively faster expansion of smaller microspheres was found to be responsible for the size effect by monitoring physicochemical changes during drug release. 　Rifampicin, a broad-spectrum antibiotic, was encapsulated by PLGA microspheres and PLGA-alginate core-shell microspheres. The long-term antimicrobial effects of drug loaded microspheres were investigated by drug release test and antimicrobial test against Staphylococcus aureus. The results showed that drug delivery devices could provide antimicrobial effect for more than one month. These drug delivery devices show potential in applications of controlled drug delivery and long-term antimicrobial therapy. 　In conclusion, drug delivery devices with different microstructures were fabricated using microfluidic method. The diameter of PLGA microspheres only influence the first phase of drug release profile (lag phase) and smaller microspheres exhibited shorter lag phase. The size effect is due to the relatively faster expansion rate of smaller microspheres. Rifampicin loaded PLGA microspheres and PLGA-alginate core-shell microspheres could provide sustained release of rifampicin for more than one month. The released rifampicin was able to inhibit the growth of Staphylococcus aureus. The controlled drug delivery devices presented showed great potential in long-term antimicrobial applications. / published_or_final_version / Orthopaedics and Traumatology / Doctoral / Doctor of Philosophy

Microfluidic droplets are a powerful tool for screening large populations of cells, molecules, and biochemical reactions. Droplet systems are able to encapsulate, incubate, screen, and sort millions of samples, providing access to large number statistics that make searching for rare events feasible. Initial development of the microfluidic devices and methods has attracted applications in biology, biochemistry, and material science, but the set of tools remains incomplete. Efforts are required to develop micro-scale droplet analogs for all bulk-scale bench top procedures and instruments. The droplet analogs must be versatile, robust, and process samples rapidly. / Engineering and Applied Sciences

The improvement of blood cell sorting techniques in recent years have attracted the attention of many researchers due to the possible benefits that these methods can lead in biology, regenerative medicine, materials science and therapeutic area. In this work a cell sorting technique based on filtration is described. The separation occurs by means of a microfluidic device, suitably designed, manufactured and tested, that is connected to an external experimental set-up. The fabrication process can be divided in two parts: at first it is described the manufacturing process of a filtering membrane, with holes of specific size that allow the passage of only certain cell types. Following the microfluidic device is fabricated through the mechanical micromilling. The membrane and the microdevice are suitably bonded and tested by means of an external connection with syringe pumps that inject blood samples at specific flow rates. The device is designed to separate blood cells and tumor cells only by using differences in size and shape. In particular during the first experiments red blood cells and platelets are sorted from white blood cells; in the other experiments red blood cells and platelets are separated from white blood cells and tumor cells. The microdevice has proven to be very efficient, in fact a capture efficiency of 99% is achieved. For this reason it could be used in identification and isolation of circulating tumor cells, a very rare cancer cell type whose presence in the bloodstream could be symptom of future solid tumor formation. The various experiments have also demonstrated that tumor cells survive even after the separation treatment, and then the suffered stress during the sorting process does not harm the biological sample.

Lymphocyte migration is crucial for adaptive immunity. CCR7 and its ligands mediate the migration and positioning of T cells in lymph nodes but the underlying mechanism is complex. The research in this thesis investigated CCR7-mediated T cell migration using a microfluidics-based approach. A microfluidic method suitable for quantitative migration analysis of genetically modified lymphocyte transfectants was developed. Using this method, I demonstrated chemotaxis of Jurkat transfectants expressing wild-type or C-terminal mutated CCR7 to a CCL19 gradient, and characterized the difference in transfectant migration mediated by wild-type and mutant CCR7. The fluorescent tag allows identification of CCR7-expressing transfectants in cell migration analysis, and microscopy assessment of CCR7 dynamics in migrating cells. Furthermore, my results also showed interesting migratory behaviours of CCR7 Jurkat transfectants in a specific co-existing CCL19 and CCL21 fields. This developed method will be broadly useful for studying cell migration signalling.

This thesis presents optical trapping of various single nanoparticles, and the method for integrating the optical trap system into a microfluidic channel to examine the trapping stiffness and to study binding at the single molecule level. Optical trapping is the capability to immobilize, move, and manipulate small objects in a gentle way. Conventional trapping methods are able to trap dielectric particles with size greater than 100 nm. Optical trapping using nanostructures has overcome this limitation so that it has been of interest to trap nanoparticles for bio-analytical studies. In particular, aperture optical trapping allows for trapping at low powers, and easy detection of the trapping events by noting abrupt jumps in the transmission intensity of the trapping beam through the aperture. Improved trapping efficiency has been achieved by changing the aperture shape from a circle; for example, to a rectangle, double nanohole (DNH), or coaxial aperture. The DNH has the advantage of a well-defined trapping region between the two cusps where the nanoholes overlap, which typically allows only single particle trapping due to steric hindrance. Trapping of 21 nm encapsulated quantum dot has been achieved which shows optical trapping can be used in technologies that seek to place a quantum dot at a specific location in a plasmonic or nanophotonic structure. The DNH has been used to trap and unfold a single protein. The high signal-to-noise ratio of 33 in monitoring single protein trapping and unfolding shows a tremendous potential for using the double nanohole as a sensor for protein binding events at a single molecule level. The DNH integrated in a microfluidic chip with flow to show that stable trapping can be achieved under reasonable flow rates of a few µL/min. With such stable trapping under flow, it is possible to envision co-trapping of proteins to study their interactions. Co-trapping is achieved for the case where we flow in a protein (bovine serum albumin – BSA) and co-trap its antibody (anti-BSA). / Graduate / 0544 / 0752 / oskuie@uvic.ca

Endothelial cells (ECs) respond to flow but the exact mechanism producing alignment is not completely understood. We characterized EC alignment in microfluidic channels, 4 mm wide by 350 um high, to generate shear of 20 dynes / cm2 across the cell surface. In microchannels, ECs aligned perpendicular under flow. Analytical tools were developed to quantify nuclear alignment at 67% for human umbilical vein endothelial cells (HUVECs); cell elongation under shear flow shifted aspect ratio from 2.41 to 2.86. We next sought to probe the mechanism through which ECs communicate during realignment. The planar cell polarity (PCP) signaling pathway is involved in cell organization and coordination during development. A number of genes are known to affect the formation and organization of cellular structures through PCP signaling in human ECs. Higher expression of Vangl1 and Dvl1 proteins did not alter cell reorganization; knockdown of Vangl1 expression decreased EC alignment.

Endothelial cells (ECs) respond to flow but the exact mechanism producing alignment is not completely understood. We characterized EC alignment in microfluidic channels, 4 mm wide by 350 um high, to generate shear of 20 dynes / cm2 across the cell surface. In microchannels, ECs aligned perpendicular under flow. Analytical tools were developed to quantify nuclear alignment at 67% for human umbilical vein endothelial cells (HUVECs); cell elongation under shear flow shifted aspect ratio from 2.41 to 2.86. We next sought to probe the mechanism through which ECs communicate during realignment. The planar cell polarity (PCP) signaling pathway is involved in cell organization and coordination during development. A number of genes are known to affect the formation and organization of cellular structures through PCP signaling in human ECs. Higher expression of Vangl1 and Dvl1 proteins did not alter cell reorganization; knockdown of Vangl1 expression decreased EC alignment.