Parallel Solid-State Electrodes for Turn-Key Intracellular Electrophysiology

用于交钥匙细胞内电生理学的并行固态电极

• 批准号: 8454608
• 负责人: Ari Chaney
• 金额: $ 31.63万
• 依托单位: STEALTH BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8454608
• 关键词: Action Potentials; Address; Alkanes; Alzheimer' s Disease; Amplifiers; Apoptotic; Area; Autistic Disorder; Axon; Basic Science; Bathing; Biochemical; Biomimetic Devices; Biomimetics; Biotechnology; Brain; Brain Diseases; Businesses; Caliber; Cell Communication; Cell Culture Techniques; Cell Survival; Cell physiology; Cells; Cellular Membrane; Characteristics; Chemicals; Collaborations; Communication; Communities; Computer software; Cultured Cells; Data; Data Quality; Deposition; Development; Devices; Disease; Electric Capacitance; Electrodes; Electronics; Electrophysiology (science); Embryo; Equipment; Esthesia; Evaluation; Feasibility Studies; Frequencies; Functional disorder; Glass; Goals; Gold; Grant; Health; Height; Hippocampus (Brain); Hour; Incubators; Industry; Injury; Ion Channel; Ions; Lead; Learning; Masks; Measurement; Measures; Medicine; Membrane; Memory; Methods; Microfabrication; Molecular; Monitor; Movement; Neurons; Neurosciences; Noise; Pathway interactions; Pattern; Performance; Personal Satisfaction; Phase; Procedures; Process; Production; Property; Protocols documentation; Puncture procedure; Rattus; Reproducibility; Research Personnel; Resistance; Resolution; Running; Scientist; Serum; Signal Transduction; Silicon; Small Business Innovation Research Grant; Specimen; Stroke; Structure; Synapses; Synaptic Transmission; System; Techniques; Technology; Testing; Thick; Tight Junctions; Time; Universities; Width; Work; base; cell injury; cell motility; cell preparation; cognitive function; cost; design; drug development; drug discovery; electric impedance; electrical measurement; electrical property; improved; interest; nanofabrication; neurophysiology; new technology; patch clamp; programs; public health relevance; relating to nervous system; response; seal; solid state; technology development; tool; voltage; voltage clamp

DESCRIPTION (provided by applicant): Interconnected networks of cells in the brain called neurons underlie all cognitive functions. Key advances in our understanding of brain function during the last several decades have resulted from technologies that permit monitoring of electrical activity in neurons. This technique, broadly referred to as electrophysiology, permits the study of circuits of connected neurons responsible for sensation, movement, thought, learning, and memory. These techniques have also revealed how abnormal electrical signaling between neurons can lead to dysfunction as occurs in disorders such as autism or Alzheimer's disease as well as due to damage from a stroke or traumatic injury. The most sensitive form of electrophysiological recording monitors very small electrical currents in single cells with glass pipettes placed inside the neuron. These 'intracellular' patch-clamp recordings are a powerful tool for exploring how neurons work-and don't work. Despite these great advances, current intracellular recording technology has significant limitations: puncturing the cell damages it, leading to short recordings and abnormal biophysical and biochemical properties; skilled scientists are required, reducing the number of labs that can use this technique; and simultaneous recording from more than one or two neurons is rarely possible, making it challenging to study neuronal communication. Stealth Biosciences was established to overcome fundamental limitations of existing intracellular techniques. Our group invented a new technology which we call "Stealth" or biomimetic electrodes. These electrodes are able to fuse into the cellular membrane, providing a minimally damaging, electrically tight junction with the cell. Our initial measurements have demonstrated high-quality, long-term intracellular recordings that rival that of traditional patch-clamps. Biomimetic probes are based on standard silicon microfabrication processing, enabling large arrays of electrodes on inexpensive chips. Using support from a Phase I SBIR grant, we will refine the device and perform feasibility studies of this game changing, inexpensive, and easy-to-use intracellular recording platform. Our long-term goals are to develop this technology for wide commercial distribution among researchers to advance basic discoveries, accelerate drug development, and improve the health and well-being of those suffering from disorders of the brain. To achieve this ambitious program we outline two Phase I Specific Aims: Aim 1: Optimize Biomimetic Electrode Performance and Production In this Aim, we will assess the performance of different geometric and architectural designs for biomimetic electrodes. Designs will be evaluated for electronic characteristics as well as for electrophysiological performance with cultured neurons. The fabrication process will be streamlined and structured with the goal of eventual large-scale fabrication. Specific milestone goals include electrical performance of < 2mV noise, better than 0.1 ms time resolution, and <200MW input impedance. Timing: Q2 and Q3. Aim 2: Functional Characterization of Biomimetic Probes with Cells in Culture The second Aim will characterize the biomimetic device performance for recording from rat hippocampal neurons. This stringent test of intracellular recording capabilities will allow direct comparison to the gold- standard pipette-based patch-clamps. Cell recordings from the different probe designs in Aim 1 will be used to optimize fabrication techniques and probe design. Long-term recordings extending for days and possibly weeks will be used to demonstrate lifetime and temporal capabilities of the probes far exceeding what is possible with conventional patch-clamps. Timing: Q3 and Q4. We have brought together a strong team with expertise in electrophysiology, micro/nanofabrication, cell-to-cell communication, and business to support this technology development at Stealth Biosciences. At the end of this program, we will have an experimentally vetted system for 'turnkey' intracellular recordings. These will provide simple cell-preparation, >16 individually addressable electrodes per chip, high-quality recordings, and compatibility with existing electrophysiological software and recording hardware. We believe these devices will find broad interest within the neuroscience community, both as a basic research tool, and for advanced applications in drug discovery and personalized medicine.

描述（由申请人提供）：大脑中称为神经元的细胞互连网络是所有认知功能的基础。过去几十年来，我们对大脑功能的理解取得了重大进展，这得益于能够监测神经元电活动的技术。这项技术被广泛地称为电生理学，可以研究负责感觉、运动、思维、学习和记忆的连接神经元的回路。这些技术还揭示了神经元之间的异常电信号如何导致功能障碍，如自闭症或阿尔茨海默病等疾病以及中风或外伤造成的损伤。最灵敏的电生理记录形式是通过放置在神经元内部的玻璃吸管来监测单个细胞中非常小的电流。这些“细胞内”膜片钳记录是探索神经元如何工作和不工作的强大工具。尽管取得了这些巨大进步，但当前的细胞内记录技术仍存在显着的局限性：刺穿细胞会对其造成损害，导致记录时间短以及生物物理和生化特性异常；需要熟练的科学家，从而减少了可以使用该技术的实验室数量；同时记录超过一两个神经元的情况几乎是不可能的，这使得研究神经元通信变得具有挑战性。 Stealth Biosciences 的成立是为了克服现有细胞内技术的基本局限性。我们小组发明了一种新技术，我们称之为“隐形”或仿生电极。这些电极能够融合到细胞膜中，与细胞提供破坏性最小、电紧密的连接。我们的初步测量表明，高质量、长期的细胞内记录可与传统膜片钳相媲美。仿生探针基于标准硅微加工工艺，可在廉价芯片上实现大型电极阵列。利用第一阶段 SBIR 拨款的支持，我们将改进该设备，并对这个改变游戏规则、廉价且易于使用的细胞内记录平台进行可行性研究。我们的长期目标是开发这项技术，以便在研究人员中进行广泛的商业推广，以推进基础发现，加速药物开发，并改善大脑疾病患者的健康和福祉。为了实现这一雄心勃勃的计划，我们概述了第一阶段的两个具体目标： 目标 1：优化仿生电极性能和生产 在这个目标中，我们将评估仿生电极的不同几何和结构设计的性能。将评估设计的电子特性以及培养神经元的电生理性能。制造过程将得到简化和结构化，以实现最终大规模制造。具体的里程碑目标包括<2mV噪声的电气性能、优于0.1ms的时间分辨率以及<200MW的输入阻抗。时间：第二季度和第三季度。目标 2：培养细胞的仿生探针的功能表征第二个目标将表征用于记录大鼠海马神经元的仿生装置的性能。这种对细胞内记录能力的严格测试将允许与基于移液管的金标准膜片钳进行直接比较。目标 1 中不同探针设计的细胞记录将用于优化制造技术和探针设计。持续数天甚至数周的长期记录将用于证明探针的寿命和时间能力远远超过传统膜片钳的能力。时间：第三季度和第四季度。我们汇集了一支在电生理学、微/纳米加工、细胞间通信和商业方面拥有专业知识的强大团队，以支持 Stealth Biosciences 的这项技术开发。在该计划结束时，我们将拥有一个经过实验审查的“交钥匙”细胞内记录系统。这些将提供简单的细胞制备、每个芯片超过 16 个可单独寻址的电极、高质量的记录以及与现有电生理软件和记录硬件的兼容性。我们相信这些设备将在神经科学界引起广泛的兴趣，既可以作为基础研究工具，也可以用于药物发现和个性化医疗的高级应用。