Nonlinear Optical Based Flow Cytometry for Purifying Stem Cell Derived Cardiomyocytes

人们对开发基于非线性光学（NLO）现象的无标记流式细胞术越来越感兴趣。这种生物光子技术，迄今为止主要用于显微镜，流式细胞仪平台的翻译需要开发新的光学方案，适合于从细胞中的NLO信号的激励流动。本项目拟研究基于非线性光学过程的无标记流式细胞仪原型的设计和开发。二次谐波产生（SHG）？通过贝塞尔光束光片几何形状与多通道微流体系统的新颖集成。此外，该SHG流式分选仪的重要性将通过解决一个具体问题来说明，即多能干细胞衍生的心肌细胞的纯化。 本项目提出研究一种新的流式细胞术方法，该方法可以在没有遗传修饰或外源标记的情况下分选细胞。尽管一些研究小组已经证明了将NLO过程应用于流式细胞术的可行性，但这些研究并没有解决与来自单个细胞的NLO信号在流过微流体系统时的一致采样相关的主要问题。该提议将提供适合于集成到流式细胞术配置中以实现可再现的细胞分析的NLO激发方案的光学设计的新进展。

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.