A scalable electrokinetic flow cytometer and cell sorter

可扩展的动电流式细胞仪和细胞分选仪

• 批准号: 10546934
• 负责人: Lorenzo Damico
• 金额: $ 35.71万
• 依托单位: AINCOBIO LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10546934
• 关键词: Address; Affect; Amplifiers; Area; Biological; Biological Markers; Biological Process; Buffers; Calibration; Cell Density; Cell Separation; Cell Size; Cell physiology; Cells; Cellular Morphology; Cellular biology; Characteristics; Charge; Communities; Complex; Construction Materials; Custom; Cytosol; Data; Devices; Electronics; Elements; Eukaryotic Cell; Field Flow Fractionation; Flow Cytometry; Fluorescence; Frequencies; Goals; Human; Immune Targeting; Immunologic Receptors; Ionic Strengths; Label; Length; Ligands; Light; Liquid substance; Maps; Measurement; Measures; Medical; Membrane; Methods; Microelectrodes; Microspheres; Morphology; Phase; Phenotype; Physics; Population; Positioning Attribute; Process; Prokaryotic Cells; Property; Research; Research Personnel; Resolution; Rest; Risk; Running; Saline; Sampling; Shapes; Signal Transduction; Small Business Innovation Research Grant; Sorting - Cell Movement; Speed; Surface; System; T-Lymphocyte; Testing; Time; Travel; Universities; Validation; Width; base; biological research; biophysical properties; cell injury; cell type; commercialization; cytotoxic CD8 T cells; density; design; digital imaging; electric field; electric impedance; electrical property; flexibility; improved; instrument; light scattering; microelectronics; microscopic imaging; novel; operation; particle; physical property; population based; prototype; receptor; scale up; simulation; voltage

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.

项目摘要 目前的细胞分离技术局限于基于细胞的异质细胞群的分选和分析。 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的表达或 受体-配体相互作用其他细胞物理学特征，如变形性、电荷， 电极化与细胞亚群之间的重要生物学差异相关， 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 基于这些细胞物理学/细胞电参数分离细胞的能力。Aincobio正在开发 “ElectroFlow”，第一台电动流式细胞仪，解决了这一差距。ElectroFlow结合介电电泳 (DEP)电场诱导力是一种公认的分离和表征 粒子，利用场流分级（FFF）基于膜极化的差异分离细胞， 其他细胞电表型。ElectroFlow将允许编程不同的力组合， 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、膜组成和形态、表面标记、表面电荷和胞质溶胶 组成）。Aincobio团队已经建立了一个原型ElectroFlow仪器，并证明了 以约5，000个细胞/分钟的速度使用真核细胞和原核细胞的概念。 Aincobio已经应用多物理模拟来重新设计设备结构和DEP 电子设备，以允许每次运行的细胞数量增加10倍。在第一阶段SBIR中，Aincobio将利用 此仿真数据用于1）构建和台架测试原型微电极阵列电路板和信号放大器 电子，和2）改善热管理，并通过测量分离效率验证原型， 活化和静息T细胞的混合物作为电压和离子强度的函数。成功完成 第一阶段的目标是建立一个商业上可行的仪器，可以分析106细胞的概念验证， 运行20分钟并通过细胞电/细胞物理学特征进行分类。ElectroFlow的商业化将提供 所有研究人员都可以使用这种方法来研究和利用细胞电特性的差异， 生物学研究和生物医学应用的新途径。