A micromachining fluidic cantilever for single cell advanced patch clamping and cellular characterization using atomic force microscopy

使用原子力显微镜进行单细胞先进膜片钳和细胞表征的微加工流体悬臂

• 批准号: 10478331
• 负责人: Ami Chand
• 金额: $ 68.04万
• 依托单位: APPLIED NANOSTRUCTURES, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10478331
• 关键词: Academia; Action Potentials; Address; Adhesives; Affect; Area; Atomic Force Microscopy; Biological; Biological Assay; Biological Markers; Biological Sciences; Biology; Biomedical Research; Cardiac; Cardiac Myocytes; Cardiology; Cardiotoxicity; Cell surface; Cells; Clinical; Consumption; Development; Devices; Disease; Doctor of Philosophy; Drug Evaluation; Elasticity; Electrodes; Electronics; Electrophysiology (science); Endocrinology; Event; Failure; Feedback; Functional disorder; Genetic; Gold; Hour; Human; In Vitro; Individual; Industry; Ion Channel; Ions; Laboratories; Laboratory Technicians; Learning; Libraries; Liquid substance; Masks; Measurement; Measures; Membrane Potentials; Microbiology; Microelectrodes; Modality; Muscle Fibers; Nanostructures; Nanotechnology; Neurology; Neurons; Neurosciences; Patients; Performance; Pharmaceutical Preparations; Pharmaceutical Solutions; Pharmacologic Substance; Pharmacology; Phenotype; Population; Property; Publishing; Pump; Reaction; Records; Research; Research Personnel; Resolution; Scanning Probe Microscopes; Scientist; Small Business Innovation Research Grant; Spectrum Analysis; Structure of beta Cell of islet; System; Techniques; Technology; Testing; Time; Tissue Sample; Toxic effect; Training; base; cantilever; consumer demand; design; drug candidate; drug discovery; elastography; electrical property; heart cell; heart function; improved; induced pluripotent stem cell; medical schools; nanomachine; novel therapeutics; patch clamp; personalized medicine; response; seal; sensor; tool; validation studies; viscoelasticity

Single patch clamping is used to multiple areas of biology such as cardiology (cardiomyocytes), neurology/neuroscience (neurons), endocrinology (pancreatic beta cells), myology (muscle fibers), and even microbiology (bacterial ion channels). Applied Nanostructures (AppNano) in partnership with the Icahn School of Medicine is bringing to the market a unique solution addressing a major market need in electrophysiology measurements. With its advanced features and unmatched resolution, the device will enable researchers in academia and in the highly competitive life sciences industry to answer important scientific questions and develop and test new drugs fueling the discovery of new pharmaceutical solutions. As a result, these companies will be better equipped to keep up with the ever-increasing consumer demand for pharmaceutical products. In this SBIR we are developing a semi-automated system based on an micro-electromechanical systems (MEMS) sensor pipette used with atomic force microscopes (AFM) that can measure, simultaneously and directly, electrophysiological properties (such as action potentials (AP)), contractile forces on single cardiomyocytes (CM), and single cell elasticity. This system offers high content analysis (HCA) at a single cell level. The system enables a significant increase in performance and a dramatic decrease in time to complete a measurement. With times <5 min compared to conventional patch clamping (2-4 hours) achieved by leveraging micromachining and advanced atomic force microscopy (force spectroscopy). The proposed system will simplify patch clamping measurements and require minimal training. This system will make it reasonably easy for any laboratory technician to conduct these measurements, in contrast to conventional patch clamping, which has a steep learning curve and requires a PhD-level scientist. In addition to action potential and contraction force, we can also evaluate the viscoelastic and adhesive properties of the cells. Our device will be capable of addressing a critical bottleneck in drug discovery that arises during the final characterization of drug candidates. The device can detect single cell changes that would otherwise be masked when averaged over large populations, offering the advantage of measuring rare events, such as toxicity indicators that affect the beating phenotype or action potential (AP) of subpopulations of CMs. This tool finds applications in: drug evaluation/discovery, in the study of Cardiomyocytes (CM) derived from human induced pluripotent stem cells (CM-iPSCs), as a general patch- clamping tool, and in clinical settings. In the setting of personalized medicine, for example, the tool allows for interrogation of enough iPSC-CM (generated from a patient’s tissue sample for instance) to produce statistically meaningful results within several minutes that would indicate an individual’s reaction to a specific drug. Additionally this tool finds application in the study to other types of cardiotoxic effects and in other fields of biomedical research that use electrophysiology (patch clamping), such as neuroscience/neurology and endocrinology.

单片夹具用于生物学的多个领域，例如心脏病学（心肌细胞）， 神经学/神经科学（神经元），内分泌学（胰腺β细胞），肌肉学（肌肉纤维），甚至 微生物学（细菌离子通道）。与伊坎学校合作的应用纳米结构（APPNANO） 医学正在为市场带来一个独特的解决方案，以解决电生理学的主要市场需求 测量。凭借其高级功能和无与伦比的分辨率，该设备将使研究人员能够 学术界和竞争激烈的生命科学行业，以回答重要的科学问题和 开发和测试新药，以发现新的药物解决方案。结果，这些公司 将有更好的能力来跟上消费者对药品的不断增长的需求。 我们正在开发基于微电力系统（MEMS）的半自动化系统（MEMS） 传感器移液器与原子力显微镜（AFM）一起使用，可以轻松，直接测量 电生理特性（例如动作电位（AP）），单个心肌细胞上的收缩力 （CM）和单细胞弹性。该系统在单个单元格级别提供高含量分析（HCA）。系统 使性能显着提高和时间急剧减少，以完成测量。和 与传统的贴片夹具（2-4小时）相比，时间<5分钟（2-4小时）通过利用微加工和 晚期原子力显微镜（力光谱）。提出的系统将简化贴片夹具 测量并需要最少的培训。该系统将使任何实验室都非常容易 与传统的贴片夹具相比，技术人员进行这些测量 学习曲线，需要博士级的科学家。除了动作潜力和收缩力，我们还可以 还评估细胞的粘弹性和粘合性特性。我们的设备将能够解决 在候选药物的最终表征期间出现的药物发现中的关键瓶颈。设备 可以检测到在大量人群上平均时否则将掩盖的单细胞更改，提供 测量罕见事件的优点，例如影响跳动表型或动作的毒性指标 CMS亚群的潜力（AP）。该工具在研究中找到：药物评估/发现中的应用 源自人诱导的多能干细胞（CM-IPSC）的心肌细胞（CM），作为一般斑块 夹紧工具，以及在临床环境中。例如，在个性化医学的情况下，该工具允许 询问足够的IPSC-CM（例如，从患者的组织样本中生成），以产生统计 有意义的结果在几分钟之内表明个人对特定药物的反应。 此外，该工具在研究中发现对其他类型的心脏毒性作用和其他领域的应用 使用电生理学（斑块夹）的生物医学研究，例如神经科学/神经病学和 内分泌学。