Josiah Rudge

BioE Ph.D. Thesis Proposal

Time and Date: 9:00 a.m., Tuesday, November 14th, 2023

Location: EBB 5029



Aniruddh Sarkar, Ph.D. (Georgia Institute of Technology)



John Blazeck, Ph.D. (Georgia Institute of Technology)

Gabe Kwong, Ph.D. (Georgia Institute of Technology)

David Myers, Ph.D. (Georgia Institute of Technology)

Fatih Sarioglu, Ph.D. (Georgia Institute of Technology)


Single-Cell Impedance Cytometry Enabling Novel Single-Cell Electroporation Capabilities in a High Throughput Device


Cell therapies have shown great promise in treating diseases such as CAR-T cells for blood cancers. Manufacturing these therapies presents considerable challenges and costs. These include quality of donor source material, transfection of cells, and controlling and measuring the quality of the product. Current therapies often transfect cells by viral vectors which are costly, have payload limitations, are difficult to target specific cells with, and present safety concerns due to immunogenicity and oncogenicity. The objective of this thesis is to create a microfluidic single-cell electroporation device and scheme that addresses these cell manufacturing concerns. Electroporation is an alternative non-viral method of transfection that is inexpensive and non-immunogenic. Additionally, it is applicable to different cell types or delivery payloads including larger payloads (e.g. CRISPR-Cas). However, electroporation can cause significant cell death, toxicity, and/or low delivery efficiencies. Here we propose to build a microfluidic device which will electronically measure properties of single cells and apply an electroporation voltage based on that measurement. This feedback can reduce cell death by tailoring the electroporation voltage for each cell. The impedance measurement itself can be used to characterize the quality of cells or distinguish between cell subtypes, all of which can alter electroporation parameters if desired. We propose to use this to develop a scheme for targeted or selective electroporation of specific cells from mixtures. We also propose scale up cell throughput to produce clinically relevant quantities of cells. Finally, we intend to verify the use of this scheme to make CAR-T cells and verify their function using in-vitro assays.