Single-Cell Trapping and Manipulations with Bipolar Electrodes

使用双极电极的单细胞捕获和操作

• 批准号: 9035392
• 负责人: Daniel T Chiu
• 金额: $ 23.18万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9035392
• 关键词: Address; Archives; Biological; Blast Cell; Cancer Relapse; Cell Line; Cell physiology; Cells; Clinical; Complex; Correlation Studies; Coupled; Cytolysis; DNA Sequence Alteration; DNMT3a; Detection; Development; Devices; Disease; Electrodes; Encapsulated; Evolution; Fluorescence; Fluorescence Microscopy; Fluorescence-Activated Cell Sorting; Future; Generations; Genes; Health; Heterogeneity; Individual; Ions; K-562; Label; Location; Microfluidics; Minority; Mutation; Myelogenous; NPM1 gene; Oils; Oxidation-Reduction; Patients; Phenotype; Population; Process; Protocols documentation; Reaction; Reagent; Refractory Disease; Research Personnel; Residual Neoplasm; Resistance; Sampling; Scheme; Shapes; Side; Silicon; Sorting - Cell Movement; System; Techniques; Technology; base; cancer cell; cancer survival; chemotherapy; electric field; fluid flow; leukemia; mutant; neoplastic cell; operation; outcome forecast; response; simulation

 DESCRIPTION: An unexpected response from a minority of cells can have a dramatic impact on the development, prognosis, and treatment of disease. For example, in the treatment of cancer, the survival and resurgence of an aggressive, subclonal lineage of the malignant cells is not exceptional. Yet, we are only just beginning to uncover these phenomena and the underlying biological mechanisms because they are obscured at the bulk scale. Detection of a rare phenotype or cellular response can require the analysis of thousands of individual cells. To address this need, we propose to develop a dielectrophoresis (DEP)-based device with a high yield of few- or single-cell capture in an array format using inexpensive and simple components. Specifically, we propose to use ion depletion and enrichment at bipolar electrodes (BPEs) to generate an array of narrow electric field maxima and minima for array-based cell capture. This technological development is significant because 1) it addresses a need for effective and inexpensive single-cell manipulation, 2) the ion enrichment and depletion zones provide strong electric field gradients (strong DEP force) having a tunable size and location, 3) the use of BPEs allows facile arraying. DEP is a versatile and powerful technique that has grown in use recently to include commercial technology. DEP can be used to transport, sort, trap, and filter cells without cell labels or expensive components and has been extended to single-cell arrayed capture. Despite these major advantages, current DEP technologies have remaining challenges to overcome, and three of the most serious shortcomings relate to the generation of the electric field gradient required for DEP force. First, while DEP technologies can be operated in parallel, there are practical barriers to achieving an array of local electric field gradients. Second, the range over which DEP force exists around these electrodes and insulating barriers can be too short for high throughput device operation. Third and finally, the size and location of the electri field produced by each of these strategies is fixed, lacking plasticity. Here, we propose to shape the electric field using localized control of the conductivity of the DEP medium via faradaic ion enrichment and depletion at an array of BPEs. The proposed technology has the following unique advantages: 1) the ion depletion and enrichment zones will generate strong DEP force around the trapping locations owing to steep electric field gradients and a synergistic effect on the complex permittivity of the DEP medium, 2) these electric field gradients can extend further from the electrodes than those generated by traditional DEP electrode arrays, 3) the trapping location can be mobilized using fluid flow.

 少数细胞的意外反应可能对疾病的发展，预后和治疗产生巨大影响。例如，在癌症的治疗中，恶性细胞的侵袭性亚克隆谱系的存活和复苏并不例外。然而，我们才刚刚开始揭示这些现象和潜在的生物学机制，因为它们在大规模上是模糊的。罕见表型或细胞反应的检测可能需要分析数千个单个细胞。为了解决这一需求，我们建议开发一种基于介电泳（DEP）的设备，该设备使用廉价且简单的组件以阵列形式具有高产量的少数或单个细胞捕获。具体而言，我们建议使用离子耗尽和富集在双极电极（BPE），以产生一个阵列的窄电场的最大值和最小值的阵列为基础的细胞捕获。这种技术发展是重要的，因为1）它解决了对有效且廉价的单细胞操作的需求，2）离子富集和耗尽区提供具有可调尺寸和位置的强电场梯度（强DEP力），3）BPE的使用允许容易的排列。DEP是一种通用且功能强大的技术，最近已在使用中发展到包括商业技术。DEP可用于运输、分选、捕获和过滤细胞，而无需细胞标签或昂贵的组件，并已扩展到单细胞阵列捕获。尽管有这些主要的优点，目前的DEP技术仍然存在需要克服的挑战，并且三个最严重的缺点涉及DEP力所需的电场梯度的产生。首先，虽然DEP技术可以并行操作，但实现局部电场梯度阵列存在实际障碍。其次，DEP力存在于这些电极和绝缘屏障周围的范围对于高通量装置操作来说可能太短。第三，也是最后一点，由这些策略中的每一个产生的电场的大小和位置都是固定的，缺乏可塑性。在这里，我们建议通过法拉第离子富集和耗尽在BPE阵列中使用DEP介质的电导率的局部控制来形成电场。所提出的技术具有以下独特优势：1）由于陡峭的电场梯度和对DEP介质的复介电常数的协同效应，离子耗尽和富集区将在捕获位置周围产生强DEP力，2）这些电场梯度可以比传统DEP电极阵列产生的电场梯度从电极延伸得更远，3）可以使用流体流动来移动捕获位置。