Portable High Throughput Digital Microfluidics and On-Chip Bacteria Cultures

An intelligent, portable, and high throughput digital microfluidic (DMF) system is developed. Chapter 1 introduces microfluidics and DMF systems. In Chapter 2, a low-cost and high resolution capacitive-to-digital converter integrated circuit is used for droplet position detection. A field-programmable gate array FPGA is used as the integrated logic hub of the system for highly reliable and efficient control of the circuit. In this chapter a fast-fabricating PCB (printed circuit board) substrate microfluidic system is proposed. Smaller actuation threshold voltages than those previously reported are obtained. Droplets (3 µL) are actuated using 200 V, 500 Hz DC pulses. Droplet positions can be detected and displayed on a PC-based 3D animation in real time. The actuators and the capacitance sensing circuits are implemented on one PCB to reduce the size of the system. In Chapter 3, an intelligent EWOD (electrowetting on dielectric) top plate control system is proposed. The dynamic top plate is controlled by a piezoelectric (PZT) cantilever structure. A high resolution laser displacement sensor is used to monitor the deflection of the top plate. The gap height optimization and the harmonic vibration significantly improve the droplet velocity and decrease the droplet minimum threshold actuation voltage. The top plate vibration induced actuation improvement is magnitude and frequency dependent. 100 µm and 200 µm vibrations are tested at 25 Hz. Vibration frequencies at 5 Hz, 10 Hz, and 20 Hz are tested while the magnitude is 200 µm. Results show greater improvements are achieved at larger vibration magnitudes and higher vibration frequencies. With a vibrated top plate, the largest reduction of the actuation voltage is 76 VRMS for a 2.0 µl DI water droplet. The maximum droplet instantaneous velocity is around 9.3 mm/s, which is almost 3 times faster than the droplet velocity without top plate vibration. Liquid that has different hysteresis such as acetonitrile with various concentrations are used as a control to show its compatibility with the proposed DMF chip. Contact line depinning under top plate vibration is observed, which indicates the underlying mechanism for the improvements in actuation velocity and threshold voltage. The top plate control technique reported in this study makes EWOD DMF chips more reliable for point of care diagnostics. In Chapter 4, the mechanisms of the improvements were investigated by observing the detailed changes in the contact angle hysteresis using both parallel and nonparallel top plates. In Chapter 5, on-chip cell cultures are used for anti-biotic resistant bacteria detection. The passively dispensed on-chip cell cultures realize the isolated micro environment electrochemistry measurement, shorten the culturing time, and reduce the required sample volume. The design of the next generation ultra-portable DMF system is covered in the Appendix. Detailed technical notes and hardware design is covered in the Appendix. The proposed portable and high throughput DMF system with on-chip cell cultures have a great potential to change the standards for micro-environment culturing technologies, which will significantly improve the efficiency of actuation, sensing, and detecting performance of the DMF systems

Modelling and control of dynamic platelet aggregation under disturbed blood flow

Diagnosis of platelet function is fundamental for identifying blood disorders of patients, assessing the impact of antiplatelet agents, and enabling the appropriate titration of individual antithrombotic treatments. Following the advancement of new technologies such as microfluidic devices and the use of control engineering methods, new devices have the potential to offer new opportunities in point-of-care diagnosis of platelet function. Such new devices may have significant utility in the development of more tailored antiplatelet therapies. The aim of this thesis is to investigate modelling and control systems which support the study of the dynamic relationship between newly discovered mechanisms of platelet aggregation and disturbed blood flow, using state-of-the-art micro-engineered technologies. In order to observe the dynamics of platelet aggregation under disturbed blood flow, blood perfusion experiments carried out on a device mimicking a scenario of severe vessel narrowing are presented. The resulting biological response, that is the aggregation of platelets, is monitored in real-time and synthesised through novel measures developed using image processing techniques. A mechanistic model identifying four distinct stages observed in the formation of the aggregate is formulated, describing the nonlinear relationship between blood flow dynamics and platelet aggregation. The observed effect of disturbed blood flow on the aggregation of platelets is then modelled mathematically employing System Identification methods. A detailed account of a novel approach for the generation of experimental data is presented, as well as the formulation of tailored mathematical model structures and the calculation of their parameters using collected data. The proposed models replicate experimental results with low variation, and the reduced number of model parameters is suggested as a novel systematic measure of platelet aggregation dynamics in the presence of blood flow disturbances. In order to stabilise, optimise, and automate the measurement of platelet function in response to disturbed blood flow, custom-made control algorithms based on principles of Sliding Mode Control and Pulse-Width Modulation are developed. Moreover, the control algorithms are developed to handle the large variability of the aggregation responses from blood types with platelet hyper- and hypo-function. Simulation results illustrate the robustness of the control algorithms in the presence of time-varying nonlinearities and model uncertainty, and indicate the possibility to regulate the extent of aggregation in the device through modulation of the blood flow rate in the microchannel. The main contribution of this thesis is the development of dynamic models and control systems that allow a systematic measurement of platelet function in response to rapid changes in the blood flow (shear rate micro-gradients), in a microfluidics device containing a scenario of disturbed blood flow. Analysis of the platelet aggregation dynamics revealed that although the aggregate growth appears to be constant at times, measuring its mean fluorescence intensity indicates an increase in the dynamics of platelet density. This densification process appears fundamental for the development of an amplification phase in the aggregation response. The proposed mathematical models and control algorithms facilitate the systematic measurement of platelet function in vitro, pioneering the development of a novel framework for automated blood disorder diagnosis