ECIS-based High Throughput Screening Instrumentation

描述（由申请人提供）：研究的长期目标是开发和制造一种仪器，可以通过高通量筛选（HTS）促进新药的发现。ECIS（electric cell-substrate impedance sensing）是由Applied BioPhysics开发的用于真实的实时研究培养细胞行为的技术。该公司生产的仪器目前在全球150多个实验室中使用，用于测量细胞单层渗透性、细胞迁移和细胞与细胞外基质相互作用等基本细胞特性。ECIS仪器仅限于低通量研究，最多可监测16个单个细胞培养物。第一阶段的研究最终在开发和制造的第一个96井版本的ECIS。该仪器仅限于测量简单的阻抗，并具有限于单个96孔板的通量。尽管有这些局限性，ECIS 9600仪器现在主要成功地销售给学术实验室。第二阶段研究的目标是建立在第一阶段的成功，并创建一个真正的高通量ECIS系统。新仪器将能够监测ECIS培养威尔斯孔在宽频谱AC频率上的复阻抗。定制电子设备将适应大量96孔板的快速数据采集，从而允许筛选数千个单独的威尔斯孔。该仪器将利用监测复阻抗的能力，并将为两个重要的基于细胞的测定量身定制。这些测定之一将涉及基于自动化的基于ECIS的伤口愈合测定的细胞迁移测量。另一个将监测细胞层屏障功能的变化。这些重要的测量目前都不能在高通量测定中完成。这些目标的实现将涉及几个步骤：“开发和测试必要的电子设备”，设计和测试专门为每个测定量身定制的负担得起的耗材阵列，“生产定制软件来运行测定和管理产生的大型数据文件”，将ECIS仪器与标准机器人板处理器连接起来。一旦制造出完整的原型仪器，我们将建立两个beta测试站点，在那里将对仪器及其软件进行彻底评估，并实施更改以产生最终的生产单元。在第三阶段，该仪器将用于制药行业的HTS检测，在寻找现代治疗药物的过程中，将对数万种测试化合物进行评估。

DESCRIPTION (provided by applicant): The long-term objective of the research is to develop and fabricate an instrument that can facilitate the discovery of new drugs via high throughput screening (HTS). ECIS (electric cell-substrate impedance sensing) is a technology developed by Applied BioPhysics to study the behavior of cultured cells in real time. Instruments manufactured by the company currently find use in over 150 laboratories world wide to measure such fundamental cell properties as permeability of cell monolayers, cell migration and cell-extra cellular matrix interactions. The ECIS instrumentation had been limited to low throughput research endeavors where a maximum of 16 individual cell cultures could be monitored. Phase I research culminated in the development and fabrication of the first 96 well version of ECIS. This instrument is limited to measurements of simple impedance and has a throughput restricted to a single 96 well plate. In spite of these limitations, the ECIS 9600 instrument is now being successfully marketed primarily to academic laboratories. The objective of the Phase II research is to build upon the success of Phase I and create a truly high throughput ECIS system. The new instrument will be capable of monitoring the complex impedance of ECIS culture wells over a broad spectrum of AC frequencies. Custom electronics will accommodate rapid data acquisition of large numbers of 96 well plates permitting thousands of individual wells to be screened. The instrument will capitalize upon the power of monitoring complex impedance and will be tailored for two important cell-based assays. One of these assays will involve measurement of cell migration based on an automated ECIS-based wound-healing assay. The other will monitor changes in barrier function of cell layers. None of these important measurements can presently be accomplished in a high throughput assay. The achievement of these goals will involve several steps: "development and testing the necessary electronics," design and testing of affordable consumable arrays specifically tailored for each assay, "production of custom software to run assays and manage the large resulting data files, "interfacing the ECIS instrumentation with standard robotic plate handlers. Once a complete prototype instrument has been fabricated, we will establish two beta test sites where the instrumentation and its software will be thoroughly evaluated and changes implemented to yield the final production unit. In Phase III, the instrument will be used in HTS assays in the pharmaceutical industry, where literally tens of thousands of test compounds will be evaluated in the search for modern therapeutic drugs.