SUMMARY

Epithelia are barrier-type cells that regulate transport into and out of the body. Dysfunction of these cells is implicated in numerous diseases such as cystic fibrosis, age-related macular degeneration, and diabetes. Since epithelial tissues perform a variety of tasks that require full confluency (e.g., secretion, absorption, and filtration), validation of tissue polarity and function are critical components of a thorough physiological exam. These experiments are commonly done electrochemically, but existing methods are some combination of (1) destructive to the cells, (2) incomplete, (3) extremely difficult, and (4) low throughput (e.g., 1-2 tissues/day). Therefore, in this proposed work, a novel tool and measurement technique will be developed to study epithelial cell function that addresses these issues. Specifically, (Aim 1) build a robot that automatically inserts a pipette into epithelia with a success rate better than a highly trained expert. (Aim 2) create a new mathematical model that enables the measurement of the – previously unobservable – apical and basolateral membrane properties using extracellular electrochemical impedance spectroscopy. (Aim 3) Design a device that can utilize the math model, derived in aim 2, to non-destructively extract apical and basolateral membrane properties and become the platform for standardized quality control in future epithelial cell therapies.