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: 6796927
• 负责人: Kevin D Gillis
• 金额: $ 56.05万
• 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/6796927
• 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定量胞吐的微型设备。膜片钳电生理和碳纤维电化学方法代表了高时间分辨率和高信息含量的胞吐分析的最新技术，但速度慢且劳动密集型。细胞群分泌的生化分析具有有限的时间分辨率，不能解决单个量子融合事件。我们将利用微芯片技术开发能够在一天内对数千个细胞进行定量胞吐分析的设备，从而大大加快基础神经科学研究的步伐。该方法还将首次实现影响神经递质胞吐的候选药物的快速和高信息量筛选。例如，用于治疗帕金森病的左旋多巴通过增加多巴胺释放量来起作用。该方法将是跨学科的，并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。