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

基本信息

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
项目类别：
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

项目摘要

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。电极化率与细胞亚群之间的重要生物学差异相关可以作为这些人群的有价值的生物标志物；然而，目前可用的仪器缺乏 Aincobio 正在开发基于这些细胞物理/细胞电参数分离细胞的能力。 “ElectroFlow”是首款动电流式细胞仪，旨在解决这一问题，ElectroFlow 结合了介电泳技术。 (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 Aincobio 团队构建了 ElectroFlow 仪器原型并进行了验证。真核和原核细胞的概念，速度约为 5,000 个细胞/分钟，以便能够缩放以适应大型细胞。细胞数量，Aincobio应用多物理场模拟来重新设计设备结构和DEP 在第一阶段 SBIR 中，Aincobio 将利用电子技术将每次运行的细胞数量扩大 10 倍。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成功完成。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 ElectroFlow 的商业化将提供 20 分钟的运行和分类。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。