Nanion Syncro Patch 96

描述（由申请人提供）：电兴奋细胞的电生理学研究，如心肌细胞、骨骼肌细胞、神经元、成骨细胞，对于从各种干细胞来源中提取这些细胞及其在再生医学、基础生物学研究和疾病研究中的应用至关重要。例如，只有电生理测量才能确定是否产生了功能性心肌细胞和神经元，因为标记表达是必要的，但不足以建立细胞表型。这些测量需要在高速（短时间常数的浴液溶液的变化）和高通量的方式下完成，以提供关于大参数空间的细胞特性的真实信息。该请求是购买SyncroPatch 96 (Nanion Technologies)，这是一种自动化千兆密封膜片钳（APC）系统，用于研究单细胞的电生理特性，具有目前可用的最高通量。通过快速生成高质量的膜片钳数据，对电兴奋细胞进行准确的药理学和生理学表征，该核心仪器将为哥伦比亚大学两个校区的七个不同的nih资助实验室提供先进的技术能力。在96孔板格式下，通过快速交换浴液（在100 ms内），在单个细胞中快速测量全剂量响应曲线。该系统将提供哥伦比亚大学医学中心、哥伦比亚大学晨边校区或纽约大都会地区任何其他大学目前都无法提供的独特功能。正如B部分对单个研究项目的详细描述，SyncroPatch 96将极大地加速我们在七个不同领域的正在进行的研究：(i)遗传性心律失常患者iPS心肌细胞的高通量筛选，（ii）确定起搏和拉伸对人iPS和胚胎干细胞（hESCs）心肌细胞成熟的影响，（iii）筛选治疗心力衰竭细胞内钙泄漏的新药，（iv）电压依赖性钾通道的调节；(v)确定关键平滑肌钾通道的结构亚基间界面，（vi）鉴定新的遗传编码Ca通道阻断剂，以及（vii）通过患者来源的iPS细胞运动神经元（iPS- mns）的小分子筛选治疗肌萎缩性侧索硬化症（ALS）的新疗法。使用SyncroPatch 96收集了拟议研究中所有五个领域的初步数据。该设备的所有主要用户都是经验丰富的NIH pi，在细胞反应分析方面具有广泛的专业知识。我们期望整个哥伦比亚大学的研究人员和纽约地区的合作者充分利用这一仪器，促进合作。

DESCRIPTION (provided by applicant): Electrophysiological studies of electrically excitable cells, such as cardiac myocytes, skeletal myoblasts, neurons, osteoblasts, are critically important for the derivation of these cells from various stem cell sources and for their applicatio in regenerative medicine, fundamental biological research, and study of disease. For example, only electrophysiological measurements can determine if functional cardiomyocytes and neurons have been generated, as marker expression is necessary but not sufficient for the establishment of cell phenotype. These measurements need to be done at high speed (short time constants for changes of bath solutions) and in high-throughput fashion to provide realistic information about the cell properties in a large parameter space. This request is to purchase a SyncroPatch 96 (Nanion Technologies), an automated giga- seal patch clamp (APC) system for studies of electrophysiological properties of single cells with the highest throughput presently available. This core instrument would provide advanced technical capabilities to seven diverse NIH-funded laboratories across both campuses of Columbia University, by rapidly generating high quality patch clamp data for accurate pharmacological and physiological characterization of electrically excitable cells. Full dose response curves are rapidly measured in individual cells with a fast exchange of the bath solutions (within 100 ms), in a 96-well plate format. The system would provide unique capabilities not presently available either at the Columbia University Medical Center, the Columbia University Morningside Campus, or at any other University in the New York Metropolitan area. As detailed in the descriptions of the individual research projects in Section B, the SyncroPatch 96 would greatly accelerate our ongoing research in seven different areas: (i) High throughput screening of iPS-cardiomyocytes derived from patients carrying heritable arrhythmias, (ii) Determination of the impact of pacing and stretch on the maturation of cardiac myocytes derived from human iPS and embryonic stem cells (hESCs), (iii) Screening for novel drugs to treat intracellular calcium leak in heart failure, (iv) Modulation of voltage-dependent potassium channels; (v) Determination of the structural inter-subunit interface in a key smooth muscle potassium channel, (vi) Identification of new genetically encoded Ca channel blockers, and (vii) Novel therapies for amyotrophic lateral sclerosis (ALS) through small molecule screens of patient-derived iPS cell motor neurons (iPS-MNs). Preliminary data were collected using the SyncroPatch 96 in all five areas of proposed research. All Major Users of this equipment are seasoned NIH PIs with extensive expertise in analysis of cellular responses. We expect the instrument to be used at its full capacity, by investigators throughout the Columbia University and collaborators in the New York City area, promoting collaboration.