Electric Impedance

Sickle Cell Disease (SCD) is a genetic disease that affects approximately 100,000 people in the USA and millions worldwide. The disease is defined by a mutation in hemoglobin, the red blood cell’s oxygen carrying component. Under hypoxic (low oxygen) conditions, the mutated hemoglobin (known as HbS) polymerizes into rigid fibers that stretch the cell into a sickle shape. These rigid cells can occlude blood vessels and cause an individual immense pain. Currently, no point-of-care devices exist in the market for assisting those with SCD. Using microfluidics with custom designed portable impedance measuring hardware we can achieve label-free in vitro analyses of SCD rheology.
This dissertation presents two impedance-based devices for finger-prick volume blood testing, including a microflow cytometer for SCD diagnostics and a vaso-occlusion tester for monitoring blood flow activities. First, the microflow cytometer is validated by measuring the electrical impedance of individual cells flowing through a narrow microfluidic channel. Cellular impedance is interpreted by changes in subcellular components due to oxygen association-dissociation of hemoglobin, using an equivalent circuit model and Multiphysics simulation. Impedance values of sickle cells exhibit remarkable deviations from normal blood cells. Such deviation is quantified by a conformity score, which allows for measurement of SCD heterogeneity, and potentially disease severity. Findings from this study demonstrate the potential for SCD screening via electrical impedance. Second, a vaso-occlusion tester is validated by measuring the impedance response of blood flow within a microfluidic mimic of capillary bed.

Electrical impedance of cells is a sensitive indicator of changes in cellular structure and biophysical characteristics. Integration of electrical impedance sensing in microfluidics can be a useful tool for characterization of blood cells for their disease state, such as sickle cell disease and malaria. The first part of this dissertation presents application of a microfluidics-based electrical impedance sensor for the study of sickle cell disease. Dynamic cell sickling-unsickling process of blood cells in response to cyclic hypoxia was measured. Strong correlation was found between the electrical impedance data and patients’ hematological parameters such as levels of sickle hemoglobin and fetal hemoglobin. In addition, application of electrical impedance spectroscopy in narrow microfluidic channel was used for label-free flow cytometry and non-invasive assay of single sickle cells under controlled oxygen level. We demonstrate the capability of this new technique in differentiating normal red blood cells from sickle cells, as well as sickled cells from unsickled cells, using normoxic and hypoxic conditions. The second part of this dissertation reports an application of electrical impedance sensing for the study of placental malaria. Testing conditions were optimized so that electrical impedance can be used for real time monitoring of different cellular and molecular level variations in this in vitro model of placental malaria. Impedance characteristics of cell proliferation, syncytial fusion and long-term response of BeWo cells to adhesion of infected erythrocytes were obtained and related to the immunostaining results and inflammatory cytokines measurements. Comparing to the conventional optical microscope-based methods, electrical impedance sensing technique can provide a label-free, real-time monitoring tool to study erythrocytes and cytoadhesion, and can further be extended to other disease models and cell types.