Prof. Ruiguo Yang
Date of this Version
Brooks, J 2020, "An Equivalent Circuit Model for Localized Electroporation On Track-Etched Membranes", master's thesis, University of Nebraska-Lincoln, Lincoln.
In vitro intracellular delivery is a fundamental challenge with broad implications across biology and medicine. There are currently no widely adopted methods capable of both delivering to millions of cells and controlling that delivery to a high degree of accuracy. One promising method is porous substrate electroporation (PSEP), where cells are grown on porous membranes and electric fields are used to permeabilize discrete portions of the cell membrane for delivery. A major obstacle to widespread use of PSEP is a poor understanding of the various impedances that constitute the system, including the impedance of the porous membrane, and how these impedances affect parameter selection for increased delivery efficiency and cell viability. To improve upon this understanding, we have developed an equivalent circuit model that closely mimics the behavior of each of the main components of the PSEP system using impedance measurements of each of these main components. This circuit model reveals for the first time the distribution of voltage across the faradaic impedances and through the channels of the porous substrate during PSEP. We also reveal the influence confluency plays on the delivery process. Furthermore, we apply various sample waveforms through our model to show how waveforms may be improved for future studies. The conclusions drawn from our model are compared to experimental data of small molecules electrophoretically driven through the membrane and from intracellular delivery of protein using PSEP. Finally, a novel PSEP system is shown with increased throughput compared to alternative PSEP systems to improve the rate at which these processes are understood and the rate at which potential applications are evaluated.
Advisor: Ruiguo Yang