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

单一膜片钳被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(胰岛β细胞)、肌学(肌纤维)，甚至 微生物学(细菌离子通道)。应用纳米结构(AppNano)与伊坎学院合作 Of Medicine正在向市场推出一种独特的解决方案，以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 学术界和竞争激烈的生命科学行业回答重要的科学问题和 开发和测试新药，推动发现新的药物解决方案。因此，这些公司 将更好地装备以跟上消费者对医药产品日益增长的需求。在……里面 我们正在开发一种基于微电子机械系统(MEMS)的半自动系统 与原子力显微镜(AFM)一起使用的传感器吸管，可以同时和直接测量， 单个心肌细胞的电生理特性(如动作电位(AP))、收缩力 (Cm)和单细胞弹性。该系统提供单细胞水平的高含量分析(HCA)。系统 可显著提高性能并大幅缩短完成测量的时间。使用 与传统的膜片钳制(2-4小时)相比，通过利用微加工和 先进的原子力显微镜(力谱学)。建议的系统将简化膜片钳制 测量和要求最低限度的培训。这一系统将使任何实验室都相当容易地 技术人员进行这些测量，与传统的膜片钳相比，它具有陡峭的 学习曲线，并需要博士水平的科学家。除了动作电位和收缩力量外，我们还可以 还要评估细胞的粘弹性和粘附性。我们的设备将能够为 在候选药物的最终表征过程中出现的药物发现的关键瓶颈。该设备 可以检测到单个细胞的变化，否则在大量人群中平均时会被屏蔽，提供 测量罕见事件的优势，例如影响跳动表型或行为的毒性指示器 不育系亚群的潜力(Ap)。该工具在以下方面有应用：药物评估/发现、研究 人诱导多能干细胞(CM-IPSCs)来源的心肌细胞(CM)作为一种通用的补片- 夹紧工具，并在临床环境中使用。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，从患者的组织样本中产生)以统计产生 在几分钟内得出有意义的结果，这将表明个人对特定药物的反应。 此外，该工具还可应用于其他类型的心脏毒性效应的研究以及 使用电生理学(膜片钳)的生物医学研究，如神经科学/神经学和 内分泌学。

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.