Development of a prototype system for assaying exocytosis from individual cells

开发用于测定单个细胞胞吐作用的原型系统

• 批准号: 8335375
• 负责人: Kevin D Gillis
• 金额: $ 31.71万
• 依托单位: EXOCYTRONICS, LLC
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-20 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8335375
• 关键词: Amplifiers; Binding; Biological Assay; Biological Sciences; Biomedical Engineering; Biomedical Research; Biosensor; Botulinum Toxins; Botulism; Businesses; Cell Adhesion Molecules; Cell Size; Cell membrane; Cell secretion; Cells; Chromaffin Cells; Clinical; Communities; Consult; Data; Detection; Development; Device Designs; Devices; Disease; Docking; Drug Delivery Systems; Drug Industry; Electrodes; Endocrine; Event; Exocytosis; FDA approved; Feedback; Fluorescence; Fluorescence Microscopy; Food Poisoning; Glass; Goals; Grant; Hormones; Hour; Housing; Image; Incubators; Individual; Industry; Label; Laboratories; Laboratory Animals; Lead; Manuals; Marketing; Measurement; Measures; Medical; Medical Research; Membrane; Methods; Microelectrodes; Microfabrication; Modification; Molecular; NIH Program Announcements; Neurons; Neurosciences Research; Neurotoxins; Neurotransmitters; Optics; Pattern; Pharmacologic Substance; Phase; Positioning Attribute; Process; Recruitment Activity; Research; Research Personnel; Resolution; Security; Signaling Molecule; Small Business Innovation Research Grant; Solutions; Support System; Surface; System; Techniques; Technology; Testing; Time; Toxin; United States National Institutes of Health; Vesicle; base; carbon fiber; experience; high throughput screening; improved; meetings; multidisciplinary; neurotransmitter release; novel; peptide hormone; prototype; research facility; research study; therapy development; user-friendly

DESCRIPTION (provided by applicant): Cells secrete hormones and neurotransmitter via exocytosis. Understanding how exocytosis is regulated and how pharmaceuticals and toxins inhibit exocytosis is of broad medical significance. The overall goal of this SBIR project is to use microfabrication technology to develop a robust and easy-to-use prototype device that can assay release of transmitter from individual cells 10- 100 fold faster and cheaper than current approaches and thereby greatly accelerate the pace of biomedical research. In addition, the devices will provide entirely novel capabilities such as the ability to simultaneously image a fluorescently labeled vesicle while measuring release from the same vesicle with an underlying transparent electrochemical electrode. The Specific Aim is to develop and test a prototype transparent multi-well electrochemical-electrode array device. Each of the wells will be a separate experimental chamber and will contain an array of electrochemical electrodes on the bottom. Individual cells will be docked to each electrode through the use of cell-sized microwells and patterning of cell-adhesion molecules. The electrodes will be transparent and patterned on a thin glass substrate to allow simultaneous high-resolution optical and electrochemical measurements. The electrode arrays will be multiplexed using a proprietary approach to simplify the number of connections required to an external amplifier. The specific milestone for Phase 1 is to record quantal exocytosis from at least 100 cells within one hour using a single multi-well device. Achieving this milestone will demonstrate that the prototype device can acquire data ~10-fold faster than the current method using carbon-fiber microelectrodes, and thus will establish that radical improvements in throughput can be achieved in a practical platform. Potential markets for the devices include the biomedical research community, the pharmaceutical industry to carry out high-throughput screening of drugs that target exocytosis, a clinical market to perform automated assays of botulism without the use of laboratory animals and a defense / homeland security market to detect neurotoxin bioweapons with cell-based biosensors. Exocytronics LLC will consult potential academic users / customers to provide feedback on device design, will obtain marketing advice from an experienced corporate partner and will obtain entrepreneurial support while housed in the MU Life Sciences Business Incubator.

性状（由申请方提供）：细胞通过胞吐作用分泌激素和神经递质。了解胞吐作用是如何调节的，以及药物和毒素如何抑制胞吐作用具有广泛的医学意义。该SBIR项目的总体目标是使用微制造技术开发一种强大且易于使用的原型设备，该设备可以比现有方法快10- 100倍且便宜地测定单个细胞释放的发射器，从而大大加快生物医学研究的步伐。此外，该设备将提供全新的能力，如同时成像荧光标记的囊泡，同时测量从同一囊泡与下面的透明电化学电极释放的能力。具体目标是开发和测试原型透明多井电化学电极阵列装置。每个威尔斯孔都是一个单独的实验室，底部有一排电化学电极。通过使用细胞大小的微孔和细胞粘附分子的图案化，将单个细胞对接到每个电极。电极将是透明的，并在薄玻璃基板上形成图案，以允许同时进行高分辨率的光学和电化学测量。电极阵列将使用专有方法进行多路复用，以简化外部放大器所需的连接数量。阶段1的具体里程碑是使用单个多孔装置在1小时内记录至少100个细胞的量子胞吐。实现这一里程碑将证明原型设备可以比使用碳纤维微电极的当前方法快10倍地获取数据，从而将确定可以在实际平台中实现吞吐量的根本改善。该设备的潜在市场包括生物医学研究界，制药行业进行靶向胞吐作用的药物的高通量筛选，临床市场进行肉毒杆菌中毒的自动检测而不使用实验室动物和国防/国土安全市场检测神经毒素生物武器基于细胞的生物传感器。Exocytronics LLC将咨询潜在的学术用户/客户，以提供有关设备设计的反馈，将从经验丰富的企业合作伙伴那里获得营销建议，并将在MU生命科学企业孵化器内获得创业支持。