A High-Throughput Microfluidic Platform for Targeted Drug Delivery and Multiplex Analysis at the Single Cancer Stem Cell Level

Abstract

Cancer stem cells (CSCs) drive tumor initiation, therapy resistance, and relapse, but traditional bulk assays and multi-well screens average heterogeneous populations and obscure the behavior of rare CSCs. This dissertation reports the design and fabrication of a high-throughput microfluidic platform that enables gradient drug delivery and multiplex analysis at single-CSC resolution. Through iterative engineering of several chip architectures, we developed a final device that integrates microstructures for deterministic single-cell capture, a microchannel network that generates stable spatial drug gradients, and a multi-electrode array compatible with fluorescence imaging and label-free electrical measurements such as impedance spectroscopy. Using model CSC populations, we demonstrate robust single-cell trapping, long-term on-chip culture, and reproducible gradient exposure, and show that the resulting single-cell datasets reveal pronounced heterogeneity in drug response within nominally identical treatment groups. This platform provides a practical foundation for future integration with high-content imaging, multi-omics analysis, and patient-derived samples.