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.

单个膜片钳用于生物学的多个领域，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（肌纤维），甚至 微生物学（细菌离子通道）。应用纳米结构（AppNano）与伊坎学校合作 为市场带来了一种独特的解决方案，可满足电生理学的主要市场需求 测量.凭借其先进的功能和无与伦比的分辨率，该设备将使研究人员能够在 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， 开发和测试新药物，推动新药物解决方案的发现。因此，这些公司 将更好地满足消费者对药品不断增长的需求。在 我们正在开发一种基于微机电系统（MEMS）的半自动系统 与原子力显微镜（AFM）一起使用的传感器移液管，可以同时和直接地测量， 电生理特性（如动作电位（AP））、单个心肌细胞上的收缩力 (CM)和单细胞弹性。该系统在单细胞水平上提供高含量分析（HCA）。系统 能够显著提高性能并显著缩短完成测量的时间。与 与利用微加工实现的传统膜片钳（2-4小时）相比， 先进的原子力显微镜（力谱）。拟议的系统将简化膜片钳 测量和需要最少的培训。该系统将使任何实验室都能相当容易地 技术人员进行这些测量，与传统的膜片钳相比，其具有陡峭的 学习曲线，需要博士水平的科学家。除了动作电位和收缩力， 还评估了细胞的粘弹性和粘附性。我们的设备将能够解决 这是药物发现中的关键瓶颈，在候选药物的最终表征期间出现。设备 可以检测单个细胞的变化，否则将被掩盖时，平均在大群体，提供 测量罕见事件的优点，例如影响搏动表型或作用的毒性指标 潜力（AP）的CM亚群。该工具应用于：药物评价/发现，研究 来自人诱导多能干细胞（CM-iPSC）的心肌细胞（CM），作为一般补丁- 夹持工具和临床环境中。例如，在个性化医疗的设置中，该工具允许 询问足够的iPSC-CM（例如从患者的组织样本产生）以统计地产生 在几分钟内得出有意义的结果，表明个体对特定药物的反应。 此外，该工具还可应用于研究其他类型的心脏毒性作用和其他领域。 使用电生理学（膜片钳）的生物医学研究，如神经科学/神经病学， 内分泌学。