Continuous-Imaging HT Screening Instrument(RMI)

描述（由申请人提供）：人类和其他几种生物体基因组的完成为将这一丰富的知识基础转化为用于改善人类医学的新产品奠定了基础。实现基因组学工作的承诺的一个关键组成部分是获得充分表征的化合物，这些化合物结合并改变特定基因产物的活性。为此，用于全基因组表达谱分析和高通量蛋白质组学的现代技术为大规模化学生物学举措提供了基础，使用化合物作为发现工具来探测生物途径，从而揭示以潜在有益或有见地的方式改变细胞表型的新蛋白质靶点。人们逐渐认识到，生物途径调控的许多关键点是基于蛋白质-蛋白质相互作用，而不是基于酶的细胞产物催化底物。因此，传统的药物发现方法在它们可以充分解决的靶点范围上是有限的。 筛选科学家发现，在生物途径调节的关键点上，细胞对化合物的反应可以通过基于图像的细胞测定来实现，该细胞测定通过模式和细胞器易位的计算机视觉自动测量蛋白质动力学。膜和细胞内受体的亚单位通常通过重组或易位来应答。此外，生理上的细胞异质性（例如，细胞分裂周期阶段特异性的）或明显随机的可以压倒整个孔的常规高通量筛选HTS读数。这些是基于细胞图像的仪器可以显著增加自动化测定的信息内容的方式的示例。基于细胞图像的筛选的缺点是，与HTS常规全孔板读取器HTS系统的100，000或更多威尔斯/天的速率相比，以中等显微镜分辨率（大约0.5 - 2.0 μ/m，具有10 - 40倍物镜）成像的仪器通常限于每天约25，000个威尔斯。虽然已经报道了更高的速率，但是这些增加通常是通过牺牲分辨率来实现的（例如，甚至更低放大率的物镜或相当大的照相机装仓）。 在这里，我们建议增加基本的图像扫描带宽（以像素/秒为单位）的10倍，目前贝克曼库尔特IC-100仪器，其原型是在博士的学术实验室开发的，并首次商业化的Q3DM公司。这一增长将通过应用全新的原理来实现，这些原理将自动对焦和图像采集并行化，以长而连续的多色条扫描载玻片和微量滴定板。这将导致使用中等分辨率物镜（10 - 40倍干放大率）的筛选速度超过每天100，000个威尔斯孔。

DESCRIPTION (provided by applicant): Completion of the genomes of humans and several other organisms creates the foundation for translating this rich knowledge base into new products for the improvement of human medicine. A critical component of realizing the promise of genomics efforts is access to well-characterized chemical compounds that bind to and alter the activities of specific gene products. To this end, modern technologies for genome-wide expression profiling and high throughput proteomics provide the enabling foundation for large-scale chemical biology initiatives, using chemical compounds as discovery tools to probe biological pathways, thereby revealing new protein targets that alter cellular phenotypes in potentially beneficial or insightful ways. It is becoming recognized that many critical points of biological pathway regulation are predicated on protein-protein interactions rather than enzyme-based catalysis of cellular products to substrates. Thus, traditional methods of drug discovery are limited in the scope of targets they can adequately address. Screening scientists are finding that elucidation of cellular responses to chemical compounds at critical points of biological pathway regulation can be enabled by image-based cellular assays that automatically measure protein dynamics via computer vision of pattern and organelle translocations. Subunits of membrane and intracellular receptors often respond by reorganization or translocation. In addition, cellular heterogeneities that are both physiological (e.g., cell division cycle phase-specific) or apparently random can overwhelm whole-well conventional high throughput screening HTS readouts. These are examples of the ways that cell-image-based instruments can dramatically increase the information content of automated assays. The drawback of cell-image-based screening has been that instruments imaging at medium microscopy resolution (approximately 0.5 - 2.0 mu/m with 10-40x objectives) are typically limited to about 25,000 wells per day as compared with rates of 100,000 or more wells/per day for HTS conventional whole-well plate-reader HTS systems. While higher rates have been reported, these increases have been typically achieved by sacrificing resolution (e.g., even lower magnification objectives or substantial camera binning). Here we propose to increase the fundamental image scanning bandwidth (measured in pixels/s) by 10- fold over the current Beckman Coulter IC-100 instrument, the prototype of which was developed in Dr. Price's academic laboratory and first commercialized by Q3DM Inc. This increase will be gained by application of fundamentally new principles that parallelize auto-focus and image acquisition to scan slides and microtiter plates in long, unbroken continuous-motion multi-color strips. This will result in screening speeds of over 100,000 wells per day using medium resolution objectives (10-40x dry magnification).