Microchip devices to assay quantal exocytosis

用于测定量子胞吐作用的微芯片装置

• 批准号: 6943069
• 负责人: Kevin D Gillis
• 金额: $ 53.65万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6943069
• 关键词: action potentials; biomedical equipment development; biotechnology; calcium; calcium ion; chemical stimulation; chromaffin cells; electrochemistry; electrophysiology; electrostimulus; exocytosis; fluid flow; high throughput technology; laboratory rat; microprocessor /microchip; miniature biomedical equipment; neuroendocrine system; photolysis

DESCRIPTION (provided by applicant): The long-term objective of our research is to develop microdevices for high-throughput measurement of quantal exocytosis from neurons and neuroendocrine cells. Patch-clamp electrophysiological and carbon-fiber electrochemical approaches represent the state-of-the-art for high time resolution and high information content assays of exocytosis but are slow and labor-intensive. Biochemical assays of secretion from cell populations have limited time resolution and can not resolve individual quantal fusion events. We will use microchip technology to develop devices that can assay quantal exocytosis from thousands of cells in a day in order to greatly accelerate the pace of basic neuroscience research. This approach will also enable, for the first time, rapid and high information content screening of drug candidates that affect exocytosis of neurotransmitter. For example, L-DOPA used to treat Parkinson's disease acts by increasing the quantal content of dopamine release. The approach will be interdisciplinary and will bring together investigators with expertise is biomedical, electrical and mechanical engineering, materials science, physics, electrochemistry, physiology and biophysics. The specific aims are: 1) Develop approaches to automatically target individual cells to electrochemical microelectrodes on microfabricated devices. 2) Develop approaches to stimulate exocytosis from cells on microdevices including rapid microfluidic solution exchange, photolysis of caged Ca and electrical stimulation of action potentials. 3) Integrate new electrochemical electrode materials into microdevices to increase sensitivity and performance. 4) Develop electronic instrumentation to allow simultaneous recording of many channels of electrochemical or electrophysiological data. The five-year goal of the project is to have actual devices on the market to serve the exocytosis research community that are at least an order of magnitude faster than current carbon-fiber approaches. Our first-year milestones for each aim are: 1) Position one or more cells at predetermined sites on a microchip. 2) Exchange extracellular solution in < 100 ms on a microchip. 3) Characterize the electrochemical properties of a diamond-like carbon microelectrode. 4) Develop inexpensive modular circuitry for basic electrochemical measurements using off-the-shelf components that can be easily scaled for approximately 12 simultaneous channels.

描述（由申请人提供）：我们研究的长期目标是开发用于高通量测量神经元和神经内分泌细胞的量子胞吐作用的微型设备。膜片钳电生理学和碳纤维电化学方法代表了胞吐作用的高时间分辨率和高信息含量测定的最新技术水平，但缓慢且劳动密集。细胞群分泌物的生化测定具有有限的时间分辨率，并且不能分辨单个量子融合事件。我们将利用微芯片技术开发一种装置，可以在一天内分析数千个细胞的量子胞吐作用，以大大加快基础神经科学研究的步伐。这种方法也将首次实现对影响神经递质胞吐作用的候选药物的快速和高信息含量筛选。例如，用于治疗帕金森病的左旋多巴通过增加多巴胺释放的量子含量来起作用。该方法将是跨学科的，将汇集具有生物医学，电气和机械工程，材料科学，物理学，电化学，生理学和生物物理学专业知识的研究人员。 具体目标是：1）开发自动将单个细胞靶向微加工装置上的电化学微电极的方法。2)开发刺激微装置上细胞胞吐的方法，包括快速微流体溶液交换，笼状Ca的光解和动作电位的电刺激。3)将新的电化学电极材料集成到微型器件中，以提高灵敏度和性能。4)开发电子仪器，允许同时记录电化学或电生理数据的多个通道。 该项目的五年目标是在市场上推出实际设备，以服务于胞吐作用研究社区，这些设备至少比目前的碳纤维方法快一个数量级。我们每个目标的第一年里程碑是：1）将一个或多个细胞定位在微芯片上的预定位置。2)在微芯片上在< 100 ms内交换细胞外溶液。3)表征类金刚石碳微电极的电化学性能。4)开发廉价的模块化电路，用于基本的电化学测量，使用现成的组件，可以很容易地扩展到大约12个同时通道。