This dissertation focuses on the applications of solid supported phospholipid membranes as mimics of the cellular membrane using lab-on-a-chip devices in order to study biochemical events such as ligand-receptor binding and the chemical mechanism for the preservation of the biomembrane. Supported lipid bilayers (SLBs) mimic the native membrane by presenting the important property of two-dimensional lateral fluidity of the individual lipid molecules within the membrane. This is the same property that allows for the reorganization of native membrane components and facilitates multivalent ligand-receptor interactions akin to immune response, cell signaling, pathogen attack and other biochemical processes. The study is divided into two main facets. The first deals with developing a novel lipopolymer supported membrane biochip consisting of Poly(ethylene glycol) (PEG)-lipopolymer incorporated membranes. The formation and characterization of the lipopolymer membranes was investigated in terms of the polymer size, concentration and molecular conformation. The lateral diffusion of the PEG-bilayers was similar to the control bilayers. The air-stability conferred to SLBs was determined to be more effective when the PEG polymer was at, or above, the onset of the mushroom-to-brush transition. The system is able to function even after dehydration for 24 hours. Ligandreceptor binding was analyzed as a function of PEG density. The PEG-lipopolymer acts as a size exclusion barrier for protein analytes in which the binding of streptavidin was unaffected whereas the binding of the much larger IgG and IgM were either partially or completely inhibited in the presence of PEG. The second area of this study presents a molecular mechanism for in vivo biopreservation by employing solid supported membranes as a model system. The molecular mechanism of how a variety of organisms are preserved during stresses such as anhydrobiosis or cryogenic conditions was investigated. We investigated the interaction of two disaccharides, trehalose and maltose with the SLBs. Trehalose was found to be the most effective in preserving the membrane, whereas maltose exhibited limited protection. Trehalose lowers the lipid phase transition temperature and spectroscopic evidence shows the intercalation of trehalose within the membrane provides the chemical and morphological stability under a stress environment.

To harness the wealth of success and computational power from the microelectronics industry, lab-on-a-chip (LOAC) applications should be fully integrated with silicon platforms. This works demonstrates a dielectrophoresis-based LOAC device built entirely on silicon using standard CMOS (complementary metal oxide semiconductor) processing techniques. The signal phases on multiple electrodes were controlled with only four electrical contacts, which connected to the device using three metal layers separated with interlayer dielectric. Indium tin oxide was deposited on a milled plastic lid to provide the conductivity and optical clarity necessary to electrically actuate the particles and observe them. The particles and medium were in the microfluidic chamber formed by using conductive glue to bond the plastic milled lid to the patterned silicon substrate. A correlation between the particle velocities and the electric field gradients was made using video microscopy and COMSOL Multiphysics ® simulations.

To harness the wealth of success and computational power from the microelectronics industry, lab-on-a-chip (LOAC) applications should be fully integrated with silicon platforms. This works demonstrates a dielectrophoresis-based LOAC device built entirely on silicon using standard CMOS (complementary metal oxide semiconductor) processing techniques. The signal phases on multiple electrodes were controlled with only four electrical contacts, which connected to the device using three metal layers separated with interlayer dielectric. Indium tin oxide was deposited on a milled plastic lid to provide the conductivity and optical clarity necessary to electrically actuate the particles and observe them. The particles and medium were in the microfluidic chamber formed by using conductive glue to bond the plastic milled lid to the patterned silicon substrate. A correlation between the particle velocities and the electric field gradients was made using video microscopy and COMSOL Multiphysics ® simulations.

Zugl.: Dortmund, Techn. Univ., Diss., 2009

The development of a novel method of droplet levitation to be employed in lab-on-a-chip (LOC) applications relies upon the mechanism of thermocapillary convection (due to the temperature dependence of surface tension) to drive a layer of lubricating gas between droplet and substrate. The fact that most droplets of interest in LOC applications are aqueous in nature, coupled with the fact that success in effecting thermocapillary transport in aqueous solutions has been limited, has led to the development of a technique for the controlled encapsulation of water droplets within a shell of inert silicone oil. These droplets can then be transported, virtually frictionlessly, resulting in ease of transport due to the lack of friction as well as improvements in sample cross-contamination prevention for multiple-use chips. Previous reports suggest that levitation of spherical O(nL)-volume droplets requires squeezing to increase the apparent contact area over which the pressure in the lubricating layer can act allowing sufficient opposition to gravity. This research explores thermocapillary levitation and translation of O(nL)-volume single-phase oil droplets; generation, capture, levitation, and translation of O(nL)-volume oil-encapsulated water droplets to demonstrate the benefits and applicability to LOC operations.

No description available.

Many applications for the analysis and processing of biological materials require the enrichment of cell subpopulations. Conventional cell sorting systems are large and expensive with complex equipment that necessitates specialized personnel for operation. Employing microfluidics technology for lab-on-a-chip adaptation of these devices provides several benefits: improved transport control, reduced sample volumes, simplicity of operation, portability, greater accessibility, and reduced cost. The designs of microfluidic cell sorting chips vary widely in literature; evaluation and optimization efforts are rarely reported. This study intends to investigate the primary components of the design to understand the effect of various parameters and to improve the performance of the microfluidic chip. Optimized individual elements are incorporated into a proposed cell sorter chip with the ability to dynamically sort target cells from a non-homogeneous solution using electrical driving forces. Numerical and experimental results are used to evaluate the sample focusing element for controlled cell dispensing, the sorting configuration for target cell collection, and the flow elements for reduced pressure effects and prevention of flow blockages. Compact models are adapted to solve the potential field and flow field in the chip and to predict the focused sample stream width. A commercial CFD package is used to perform 2-D simulations of the potential, velocity, and concentration fields. A fluorescence microscopy visualization system is implemented to conduct experiments on several generations of chip designs. The data from sample focusing experiments, performed with fluorescent dye samples, is analyzed using a Gaussian distribution model proposed in this work. A technique for real-time monitoring of fluorescent microspheres in the microfluidic chip enables the use of dynamic cell sorting to emulate fully autonomous operation. The performance values obtained from these experiments are used to characterize the various design configurations. Sample focusing is shown to depend largely on the relative size of the sheath fluid channel and the sample channel, but is virtually independent of the junction shape. Savings in the applied potential can be achieved by utilizing the size dependency. The focusing performance also provides information for optimizing the widths of the channels relative to the cell size. Successful sorting of desired cells is demonstrated for several designs. Key parameters that affect the sorting performance are discussed; a design employing the use of supplemental fluid streams to direct the particle during collection is chosen due to a high sorting evaluation and a low sensitivity to flow anomalies. The necessary reduction of pressure influences to achieve reliable flow conditions is accomplished by introducing channel constrictions to increase the hydrodynamic resistance. Also, prolonged operation is realized by including particle filters in the proposed design to prevent blockages caused by the accumulation of larger particles. A greater understanding of the behaviour of various components is demonstrated and a design is presented that incorporates the elements with the best performance. The capability of the microfluidic chip is summarized based on experimental results of the tested designs and theoretical cell sorting relationships. Adaptation of this chip to a stand-alone, autonomous device can be accomplished by integrating an optical detection system and further miniaturization of the critical components.

No description available.

Microdroplet microfluidics has gained much interested in the past decade due to its ability to conduct a wide variety of biological and microfluidic experiments with extremely high repeatability on a mass scale. In particular the ability to culture multiple batches of cells by creating microdroplets with a single encapsulated cell and observe their growth overtime allows for specific conditioning of cells. In addition, when conducting co-culture experiment the induction of a certain stimulus may provide observational rare differences in growth that may be characterized by harnessing a single batch of cells out of thousands of samples. This thesis first presents a variety of microdroplet microfluidic devices that use specific techniques to sufficiently produce, synchronize, merge, and analyze microdroplets. Although many of the devices are capable of producing stable droplets and somewhat efficient synchronization, the overall merging efficiency for most passive or active merging methods alone is lacking. Improvements on such methods and the incorporation of multiple merging methods can lead to a higher overall merging efficiency and greater droplet stability. Also, multiple droplet detection methods can be employed to analyze cellular growth under different conditions, while passive or active sorting methods can be used to acquire particular microdroplet samples downstream. The work presented in this thesis entails the characterization and detailed analysis of all aspects of microdroplet microfluidics necessary to adequately produce a microdroplet co-culture device for microbial studies. This includes the incorporation of multiple microdroplet generators for the production of water droplets immersed in oil serving as bio-reactors for cell culture experiments. In addition, multiple microdroplet synchronization devices were tested to sufficiently align multiple trains of droplets for downstream merging using a variety of passive, active, or combination merging methods. In particular, the use of an electric field can cause destabilization of the surfactant surrounding a microdroplet and allow for the formation of a liquid bridge. The formation of this liquid bridge in conjunction with passive merging methods can lead to droplet electrocoalescence. The incorporation of a more uniform electric field that reduces the angle between the droplet dipole moment and E-field can lead to better droplet merging while reducing voltage and frequency requirements observed in previously publications. The testing, observation, and optimization of such aspects of microdroplet microfluidics are crucial for the advancement and production of sound microdroplet culture devices for a variety of applications including the analysis of dangerous pathogenic substances, drug testing or delivery, and genetic studies.

Continuous progress in the nanotechnology field has allowed for the emergence of powerful, nanopore-based detection technology. Solid-state nanopores were developed for next-generation sequencing and single-molecule detection. They are advantageous over their biological counterpart because they offer robustness, stability, tunable pore size and the ability to be integrated within a microfluidic device. With all of these attractive attributes, solid-state nanopores are a top contender for point-of-care diagnostic technologies. However, hindering their performance is an inability to distinguish between small molecules, pore-clogging, and the detection rate's dependence on sample concentration. The concentration-dependent detection rate becomes particularly evident at low sample concentrations (<1 nM), sometimes taking hours for the nanopore to sense a single molecule because of diffusion. The inability to distinguish between small molecules can be addressed using DNA nanostructures; however, pore-clogging and variable detection rates hinder its potential in a clinical setting. This thesis proposes a microfluidic device design and methodology that seeks to mitigate pore-clogging and improve the detection rate for dilute samples. DNA coated microbeads will create a bead column within the microfluidic device and confine the target molecules to an extremely small (20 nL) volume. The sample can be washed, ridding the contaminants, and eluted on-chip, so the sample is purified and concentrated, affording a more reliable sensing performance. First, a magnetic microbead DNA assay was optimized off-chip, and the capture and release efficiencies were monitored using a Biotek™ Epoch™ 2 spectrophotometer (Chapter 2). Next, a novel microfluidic device design was optimized and validated to ensure precise sample manipulation (Chapter 3). Finally, the microbead assay was incorporated into the microfluidic device for sample concentration (Chapter 4). Fluorescence microscopy results suggest successful DNA elution from the microbeads within the microfluidic device, allowing for a 28.5 X concentration increase. This platform shows promise for sample preconcentration by reducing the starting DNA sample volume of 25 µL to 20 nL, which could improve the speed of solid-state nanopore sensing.