Nonlinear Optical Based Flow Cytometry for Purifying Stem Cell Derived Cardiomyocytes

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1264776 - ChanThere has been a growing interest in developing label-free flow cytometry based on nonlinear optical (NLO) phenomenon. The translation of such biophotonic techniques, which thus far have been primarily used in microscopy, to a flow cytometry platform requires the development of novel optical schemes appropriate for the excitation of NLO signals from cells in flow. This project proposes to study the design and development of a prototype label-free flow cytometer based on a NLO process ? second harmonic generation (SHG) ? by the novel integration of a Bessel beam lightsheet geometry with multichannel microfluidic systems. In addition, the importance of this SHG flow sorter will be illustrated by addressing a specific problem, the purification of pluripotent stem cell derived cardiomyocytes. This project proposes to study a new approach for flow cytometry that can sort cells without genetic modification or exogenous labeling. Although a few groups have demonstrated the feasibility of implementing NLO processes to flow cytometry, those studies did not address major issues related to the consistent sampling of NLO signals from individual cells as they flow through the microfluidic system. This proposal will provide new advancements in the optical design of the NLO excitation scheme appropriate for integration into a flow cytometric configuration to achieve reproducible cellular analysis.