A scalable electrokinetic flow cytometer and cell sorter

项目总结 目前的细胞分离技术仅限于分类和分析基于 细胞大小或细胞密度的差异，或通过利用特定免疫靶点或 受体-配体相互作用。其他细胞物理特性，如可变形性、电荷和 电极化率与细胞亚群和细胞亚群之间的重要生物学差异有关 将作为这些人群的有价值的生物标志物；然而，目前可用的工具缺乏 根据这些细胞物理/细胞电学参数分离细胞的能力。阿因科比奥正在开发 “ElectrFlow”，第一台电动流式细胞仪，解决了这一空白。电流法结合介电泳法 (DEP)，一种电场感生的力，被公认为一种分离和表征 颗粒，场流动分级(FFF)，根据膜极化和细胞的差异分离细胞 其他细胞电学表型。ElectrFlow将允许对不同的力组合进行编程 不同的细胞类型或细胞的不同方面，实现细胞特征(大小， 形状、密度、膜成分和形态、表面标志、表面电荷和胞浆 构图)。Aincobio团队已经建造了一个电流仪的原型，并展示了 概念与真核和原核细胞都以~5,000个/分钟的速度进行。支持扩展以适应大型 细胞的数量，Aincobio应用了多物理模拟来重新设计设备结构和DEP 电子设备允许每次运行的电池数量扩大10倍。在第一阶段SBIR中，Aincobio将利用 该仿真数据用于1)搭建微电极阵列电路板和信号放大器样机并进行台架测试 2)改进热管理，并通过测量分离效率来验证样机 作为电压和离子强度的函数的激活和静止T细胞的混合物。圆满完成 第一阶段AIMS将为商业上可行的仪器建立概念验证，该仪器可以分析106细胞 一次20分钟的跑步，根据细胞电/细胞物理特征进行分类。ElectrFlow的商业化将提供 便于所有研究人员研究和利用细胞电学特性差异的手段，完全开放 生物研究和生物医学应用的新途径。

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.