A scalable electrokinetic flow cytometer and cell sorter

PROJECT SUMMARY Current cell separation technologies are limited to sorting and analyzing heterogenous cell populations based on differences in cell size or cell density or by leveraging the expression of specific immunological targets or receptor-ligand interactions. Other cytophysical characteristics such as deformability, electrical charge, and electrical polarizability are correlated with important biological differences between subpopulations of cells and would serve as valuable biomarkers for these populations; however, currently available instruments lack the capability to separate cells based on these cytophysical/cytoelectrical parameters. Aincobio is developing “ElectroFlow”, the first electrokinetic flow cytometer, to address this gap. ElectroFlow combines dielectrophoresis (DEP), an electrical-field-induced force that is well-established as a method to separate and characterize particles, with field-flow-fractionation (FFF) to separate cells based on differences in membrane polarization and other cytoelectric phenotypes. ElectroFlow will allow for different force combinations to be programmed for different cell types or different aspects of cells, achieving specialized differentiation of cellular features (size, shape, density, membrane composition and morphology, surface markers, surface charge, and cytosol composition). The Aincobio team has built a prototype ElectroFlow instrument and demonstrated proof-of- concept with both eukaryotic and prokaryotic cells at ~5,000 cells/min. To enable scaling to accommodate large numbers of cells, Aincobio has applied multi-physics simulations to redesign the device construction and DEP electronics to allow for a 10x scale-up in number of cells per run. In this Phase I SBIR, Aincobio will leverage this simulation data to 1) build and bench-test prototype microelectrode array circuit boards and signal amplifier electronics, and 2) improve thermal management and validate prototype by measuring separation efficiency of mixtures of activated and resting T-cells as a function of voltage and ionic strength. Successful completion of the Phase I aims will establish proof-of-concept for a commercially viable instrument that can analyze  106 cells in a 20-minute run and sort by cytoelectric/cytophysical features. Commercialization of ElectroFlow will provide an accessible means for all researchers to study and harness differences in cytoelectric properties, opening entirely new avenues of biological research and biomedical application.