Carbon Nanopipette ElectroProbe in Cell-Electrophysiology Applications

碳纳米移液器电探针在细胞电生理学应用中的应用

• 批准号: 7746216
• 负责人: Chamika Wansapura
• 金额: $ 20.02万
• 依托单位: VEGRANDIS, LLC
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7746216
• 关键词: Affect; Area; Automation; Biology; Blood capillaries; Caliber; Carbon; Cell Count; Cell Extracts; Cell Survival; Cell membrane; Cells; Characteristics; Chemical Stimulation; Chemicals; Classification; Data; Development; Devices; Diagnostic; Dimensions; Distal; Drug Delivery Systems; Electrodes; Electrolytes; Electron Microscopy; Electrons; Electrophysiology (science); Environment; Feasibility Studies; Film; Future; Generations; Glass; Goals; Individual; Injection of therapeutic agent; Laboratories; Left; Legal patent; Length; Letters; Licensing; Liquid substance; Location; Marketing; Measurement; Measures; Mechanics; Membrane Potentials; Methods; Microinjections; Modification; Molecular; Needles; Nucleic Acids; Organelles; Pattern; Pattern Recognition Systems; Penetration; Pennsylvania; Phase; Physiology; Positioning Attribute; Price; Process; Production; Productivity; Property; Proteins; Quality Control; Quartz; Reagent; Reproducibility; Research; Research Personnel; Scanning; Signal Transduction; Small Business Innovation Research Grant; Suction; Surface; Survival Rate; System; Technology; Testing; Time; Tube; Universities; Variant; Work; base; capillary; cellular engineering; cost; design; electrical measurement; electrical potential; extracellular; improved; innovative technologies; macromolecule; manufacturing process; nanometer; nanosized; novel; prototype; public health relevance; research study; response; tool; voltage; voltage clamp

DESCRIPTION (provided by applicant): The main goal of this Phase I SBIR project is to optimize the fabrication process and demonstrate the feasibility of using a novel carbon-based nanopipette (CNP electroprobe) for penetration of cell membrane in single cells with minimal cell intrusion and allow measurement of changes in potential or ionic currents across the cell membrane. CNP fabrication does not require any cumbersome assembly. The CNP consists of a pulled glass capillary terminating with an exposed carbon pipe with a diameter ranging from tens to hundreds of nanometers. The entire inner lumen of the glass capillary is coated with a carbon film that provides an electrically conducting path from the distal end of the capillary to the exposed carbon tip. Conducting films can be patterned on the glass capillary's outer surface to form counter/reference electrodes. These devices can inject chemicals and macromolecules into single cells, extract fluids and specific proteins from cell's interior, and generate electrical signals upon cell penetration. The CNPs offer significant advantages over the commonly used pulled glass pipette electrodes such as smaller tip size (minimal damage to cells and the ability to probe organelles), better mechanical properties, higher durability (tips do not break or clog easily), potential for automation (the cell's penetration can be sensed through an electric signal), potential to carry out electrophysiological measurements concurrently with injection, and multifunctional analytic capabilities while being competitive in price with the glass micropipettes. Moreover, given their durability, the CNP electroprobe offers higher efficiency and lower cost (on a per cell basis) than their glass counterparts. Unlike their glass counterparts, the CNPs do not require electrolyte inside the capillary tube due to the carbon tip is in contact with cellular medium. In addition to optimizing fabrication process to refine probe tips, in this proposal, the unique characteristic of CNPs as nanoelectrodes will be demonstrated by measuring the variations in change in membrane potential, or change in ionic currents by using CNP nanoelectrodes for recording inside a single cell. PUBLIC HEALTH RELEVANCE: Demonstration of CNP electrodes to penetrate single cells with minimum damage to cell membrane and allow reliable and reproducible measurements of changes in membrane potential or total ionic currents will allow advancement in electrophysiology applications of single cells by replacing the conventional glass micropipette electrodes with CNP electroprobes. The proposed CNP devices will have applications in manipulation and study of individual cells in fields of biology, physiology, biomedical diagnostics, and in drug delivery.

描述（由申请方提供）：该I期SBIR项目的主要目标是优化制造工艺，并证明使用新型碳基纳米移液管（CNP电探针）穿透单细胞中的细胞膜的可行性，细胞侵入最小，并允许测量跨细胞膜的电位或离子电流的变化。CNP制造不需要任何繁琐的组装。CNP由一个拉伸的玻璃毛细管组成，其末端是一个直径范围从几十到几百纳米的暴露碳管。玻璃毛细管的整个内腔涂有碳膜，该碳膜提供从毛细管远端到暴露的碳尖端的导电路径。导电膜可以在玻璃毛细管的外表面上图案化以形成对电极/参比电极。这些装置可以将化学物质和大分子注入单个细胞，从细胞内部提取液体和特定蛋白质，并在细胞穿透时产生电信号。 与常用的拉丝玻璃移液管电极相比，CNP具有显著优势，例如尖端尺寸较小（对细胞的损伤最小，探测细胞器的能力），更好的机械性能，更高的耐久性（尖端不易断裂或堵塞），自动化潜力（可以通过电信号感测细胞的穿透），与注射同时进行电生理学测量的潜力，和多功能分析能力，同时在价格上与玻璃微量移液器竞争。此外，由于其耐用性，CNP电探针提供了比玻璃对应物更高的效率和更低的成本（以每个电池为基础）。与其玻璃对应物不同，CNP不需要毛细管内的电解质，因为碳尖端与细胞培养基接触。除了优化制造工艺以改进探针尖端之外，在该提案中，CNP作为纳米电极的独特特性将通过使用CNP纳米电极在单个细胞内记录来测量膜电位变化或离子电流变化的变化来证明。 公共卫生相关性：证明CNP电极可以穿透单细胞，对细胞膜的损伤最小，并可以可靠且可重复地测量膜电位或总离子电流的变化，这将通过用CNP电探针取代传统的玻璃微量移液器电极来推进单细胞的电生理学应用。提出的CNP装置将在生物学、生理学、生物医学诊断和药物递送领域中的单个细胞的操纵和研究中具有应用。