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.

项目概要 目前的细胞分离技术仅限于分选和分析异质细胞群 细胞大小或细胞密度的差异或通过利用特定免疫靶标的表达或 受体-配体相互作用。其他细胞物理特征，例如变形性、电荷和 电极化率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/细胞电参数分离细胞的能力。安科生物正在研发 “ElectroFlow”是第一台动电流式细胞仪，旨在解决这一空白。 ElectroFlow 结合介电泳 (DEP)，一种电场感应力，是一种成熟的分离和表征方法 颗粒，通过场流分级 (FFF) 根据膜极化和分离的差异来分离细胞 其他细胞电表型。 ElectroFlow 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、膜组成和形态、表面标记、表面电荷和胞质溶胶 作品）。 Aincobio 团队构建了 ElectroFlow 仪器原型并进行了证明- 真核和原核细胞的概念，速度约为 5,000 个细胞/分钟。为了能够缩放以适应大 细胞数量，Aincobio应用多物理场模拟来重新设计设备结构和DEP 电子设备允许每次运行的细胞数量扩大 10 倍。在第一阶段 SBIR 中，Aincobio 将利用 该仿真数据用于 1) 构建和台架测试原型微电极阵列电路板和信号放大器 电子设备，2) 通过测量分离效率来改进热管理并验证原型 激活和静息 T 细胞的混合物作为电压和离子强度的函数。圆满完成了 第一阶段的目标是为商业上可行的仪器建立概念验证，该仪器可以分析 106 个细胞 20 分钟运行并按细胞电/细胞物理特征分类。 ElectroFlow 的商业化将提供 为所有研究人员提供研究和利用细胞电特性差异的可用方法，完全开放 生物研究和生物医学应用的新途径。