Bislug flow in circular and noncircular channels and the role of interface stretching on energy dissipation

The area of microfluidics has increased in popularity with such fields as MEMS, microreactors, microscaleheat exchangers, etc. A comprehensive understanding of dissipation mechanisms for fluid flow in microchannels is required to accurately predict the behavior in these small systems. Tests were performed using a constant pressure potential created by two immiscible fluids juxtaposed in a microchannel. This study focused on the flow and dissipation mechanisms in round and square microchannels. There are four major dissipation mechanisms in slug flow; wall shear, dissipation at the contact line, menisci interaction and the stretching of the interface. A force balance between the internal driving potential, viscous drag and interface stretching was used to develop a model for the prediction of the velocity of a bislug in a microchannel. Interface stretching is a dissipation mechanism that has been included due to the unique system properties and becomes increasingly more important as the bislug decreases in length.

Non-intrusive pressure measurement in microchannels

A non-intrusive interferometric measurement technique has been successfully developed to measure fluid compressibility in both gas and liquid phases via refractive index (RI) changes. The technique, consisting of an unfocused laser beam impinging a glass channel, can be used to separate and quantify cell deflection, fluid flow rates, and pressure variations in microchannels. Currently in fields such as microfluidics, pressure and flow rate measurement devices are orders of magnitude larger than the channel cross-sections making direct pressure and fluid flow rate measurements impossible. Due to the non-intrusive nature of this technique, such measurements are now possible, opening the door for a myriad of new scientific research and experimentation. This technique, adapted from the concept of Micro Interferometric Backscatter Detection (MIBD), boasts the ability to provide comparable sensitivities in a variety of channel types and provides quantification capability not previously demonstrated in backscatter detection techniques. Measurement sensitivity depends heavily on experimental parameters such as beam impingement angle, fluid volume, photodetector sensitivity, and a channel’s dimensional tolerances. The current apparatus readily quantifies fluid RI changes of 10-5 refractive index units (RIU) corresponding to pressures of approximately 14 psi and 1 psi in water and air, respectively. MIBD reports detection capability as low as 10-9 RIU and the newly adapted technique has the potential to meet and exceed this limit providing quantification in the place of detection. Specific device sensitivities are discussed and suggestions are provided on how the technique may be refined to provide optimal quantification capabilities based on experimental conditions.

EFFECTS OF ION LIMITING CONDITIONS ON THE BEHAVIOR OF MICROFLUIDIC DEVICES

Finite numbers of ions are present in microfluidic devices. This leads to ion limiting effects in microfluidic channels and electrode surfaces. These effects include electrode surface changes and ion concentration gradient formation across microfluidic channels, and can influence microfluidic device behavior. A literature survey on the use of electrochemical analysis techniques in micro- and nanofluidic devices was carried out, which puts into perspective the importance of electrode surface changes with regards to analytical microfluidic applications. Surface changes in Pt wire electrodes under various physiological buffer and electric field conditions were investigated using cyclic voltammetry (CV), SEM-EDS and XPS. Effects of surface changes on electrochemical analysis performance of Pt wire and thin film electrodes were investigated. Electrode surfaces were subjected to varying phosphate buffer and electric field conditions, and their CV performance was monitored. Electrode surfaces were also studied with SEM-EDS. Two studies of ion concentration gradient formation in microfluidic channels were conducted. In the first, concentration gradients of H+ and OH- ions generated on electrode surfaces were found to cause significant pH decreases in certain buffer and electric field conditions, which was also found to play a key role in iDEP manipulation of proteins. The role of electrode surface reactions in this case shows the importance of understanding electrode surface changes in microfluidic devices. In the second study of ion concentration gradient formation, Cl- ion concentration gradient formation was attempted to be quantified upon electric field application across a KCl solution. Electrokinetic transport of the Cl- indicating fluorophore MQAE contributed significantly to the fluorescence microscopy signals collected, complicating Cl- quantification as a function of position and time. It was shown that a dielectric coating on electrode surfaces is effective at preventing MQAE electrokinetic transport.

ALTERNATING CURRENT DIELECTROPHORETIC MANIPULATION OF ERYTHROCYTES IN MEDICAL MICRODEVICE TECHNOLOGY

Medical microdevices have gained popularity in the past few decades because they allow the medical laboratory to be taken out into the field and for disease diagnostics to happen with a smaller sample volume, at a lower cost and much faster. Blood is the human body's most readily available and informative diagnostic fluid because of the wealth of information it provides about the body's general health including enzymatic, proteomic and immunological states. The purpose of this project is to optimize operating conditions and study ABO-Rh erythrocytes dielectrophoretic responses to alternating current electric signals. The end goal of this project is the creation of a relatively inexpensive microfluidic device, which can be used for the ABO-Rh typing of a blood sample. This dissertation presents results showing how blood samples of a known ABO- Rh blood type exhibit differing behavior to the same electrical stimulus based on their blood type. The first panel of donors and experiments, presented in Chapter 4 occurred when a sample of known blood type was injected into a microdevice with a T-shaped electrode configuration and the erythorcytes were found to rupture at a rate specific to their ABO-Rh blood type. The second set of experiments, presented in Chapter 5, were originally published in Electrophoresis in 20111. Novel in this work was the discovery that treatment of human erythrocytes with β-galactosidase successfully removed ABO surface antigens such that native A and B blood no longer agglutinated with the proper antibodies. This work was performed in a medium of conductivity 0.9S/m which is close to the measured conductivity of pooled plasma (~1.1S/m). The ability to perform dielectrophoresis experiments at physiological conductivities conditions is advantageous for future portable devices because the device/instrument would not need to store dilution buffers. The final results of this project, presented in Chapter 6, explore the entire dielectrophoretic spectra of the ABO-Rh erythrocytes including the cross-over frequency and the magnitudes of the positive or negative dielectrophoretic response. These were completed at lower medium conductivities of 0.1S/m and 0.01-0.04S/m. These results show that by using the sweep function built into the Agilent alternating current generator it is possible to explore how a single group of blood cells will react to rapid changes in frequency and will provide the user with curve that can be matched the theoretical dielectrophoretic response curves. As a whole this project shows that it is possible to distinguish human erythrocytes by their ABO-Rh blood type via three different dielectrophoretic methods. This work builds on the foundation of that it is possible to distinguish healthy from infected cells2-7, similar cell types1,7-14 and other work regarding the dielectrophoresis of human erythrocytes1,10,11. This work has implications in both medical diagnostics and future dielectrophoretic work because it has shown that ABO-Rh blood type is now a factor, which must be identified when working with a human blood sample. It also shows that the creation of a microfluidic device that subjects human erythrocytes to a dielectrophoretic impulse and then exports an ABO-Rh blood type is a near future possibility.

DEVELOPMENT OF CONFOCAL IMAGING TECHNIQUES FOR PROBING INTERFACIAL DYNAMICS IN MICROSCALE, GAS-LIQUID, TWO-PHASE FLOW

Micro-scale, two-phase flow is found in a variety of devices such as Lab-on-a-chip, bio-chips, micro-heat exchangers, and fuel cells. Knowledge of the fluid behavior near the dynamic gas-liquid interface is required for developing accurate predictive models. Light is distorted near a curved gas-liquid interface preventing accurate measurement of interfacial shape and internal liquid velocities. This research focused on the development of experimental methods designed to isolate and probe dynamic liquid films and measure velocity fields near a moving gas-liquid interface. A high-speed, reflectance, swept-field confocal (RSFC) imaging system was developed for imaging near curved surfaces. Experimental studies of dynamic gas-liquid interface of micro-scale, two-phase flow were conducted in three phases. Dynamic liquid film thicknesses of segmented, two-phase flow were measured using the RSFC and compared to a classic film thickness deposition model. Flow fields near a steadily moving meniscus were measured using RSFC and particle tracking velocimetry. The RSFC provided high speed imaging near the menisci without distortion caused the gas-liquid interface. Finally, interfacial morphology for internal two-phase flow and droplet evaporation were measured using interferograms produced by the RSFC imaging technique. Each technique can be used independently or simultaneously when.

UTILIZING DIELECTROPHORESIS TO DETERMINE THE PHYSIOLOGICAL DIFFERENCES OF EUKARYOTIC CELLS

Type 1 diabetes affects over 108,000 children, and this number is steadily increasing. Current insulin therapies help manage the disease but are not a cure. Over a child’s lifetime they can develop kidney disease, blindness, cardiovascular disease and many other issues due to the complications of type 1 diabetes. This autoimmune disease destroys beta cells located in the pancreas, which are used to regulate glucose levels in the body. Because there is no cure and many children are affected by the disease there is a need for alternative therapeutic options that can lead to a cure. Human mesenchymal stem cells (hMSCs) are an important cell source for stem cell therapeutics due to their differentiation capacity, self-renewal, and trophic activity. hMSCs are readily available in the bone marrow, and act as an internal repair system within the body, and they have been shown to differentiate into insulin producing cells. However, after isolation hMSCs are a heterogeneous cell population, which requires secondary processing. To resolve the heterogeneity issue hMSCs are separated using fluorescent- and magnetic-activate cell sorting with antigen labeling. These techniques are efficient but reduce cell viability after separation due to the cell labeling. Therefore, to make hMSCs more readily available for type 1 diabetes therapeutics, they should be separated without diminishing there functional capabilities. Dielectrophoresis is an alternative separation technique that has the capability to separated hMSCs. This dissertation uses dielectrophoresis to characterize the dielectric properties of hMSCs. The goal is to use hMSCs dielectric signature as a separation criteria rather than the antigen labeling implemented with FACS and MACS. DEP has been used to characterize other cell systems, and is a viable separation technique for hMSCs.