PFI-TT: Throughput Characterization of a Prototype Second Harmonic Generation Flow Cytometer for Stem Cell Derived Cardiomyocyte Purification

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The broader impact/commercial potential of this PFI project is the development of a new technology for purifying stem cell derived cardiomyocytes (SC-CMs). SC-CMs are a promising source of heart cells that can be grown in the laboratory from stem cells and used to address significant healthcare issues facing this country. These cells can potentially be used to engineer tissue to treat patients with heart disease/failure, or to screen for cardiotoxic side effects of drugs. Before these cells can be used for these applications, however, the cells must be purified to remove undesired cells from the cell population that may affect normal heart function. Currently, no established technique can reliably perform this purification. The project proposes to develop a new optical technique that can specifically identify SC-CMs and to create a prototype instrument that can efficiently perform this analysis. The long-term impact of this project will be to improve healthcare and quality of life of patients by being able to use these purified cells to treat patients with heart disease and to improve the drug discovery and development process. The proposed project focuses on developing a prototype second harmonic generation (SHG) flow cytometer to detect and identify SC-CMs without the use of exogenous labels. The research goals and objectives are to 1) design and integrate an optical system for SHG signal detection with microfluidic systems commonly used in flow cytometry, 2) optimize the system and characterize the maximum theoretical analytical throughput, and 3) assess cell viability after the analysis. A lightsheet optical system will be developed based on scanned Bessel and elongated Gaussian beams, and combined with microfluidic cell handling systems. Studies will be performed to evaluate the accuracy for identifying SC-CMs and characterizing the throughput of the system. Theoretical estimates of maximum throughput will be determined by extrapolation of signal strength and flow rate data. Cell viability post-analysis will be assessed using standard cell assays.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.