Continuous-Imaging HT Screening Instrument(RMI)

描述(申请人提供)：人类和其他几种生物基因组的完成为将这一丰富的知识库转化为改进人类医学的新产品奠定了基础。实现基因组学努力前景的一个关键组成部分是获得具有良好特征的化合物，这些化合物与特定基因产物结合并改变其活性。为此，全基因组表达谱和高通量蛋白质组学的现代技术为大规模的化学生物学倡议提供了使能的基础，使用化合物作为发现工具来探索生物途径，从而揭示以潜在有益或有洞察力的方式改变细胞表型的新的蛋白质靶标。人们逐渐认识到，许多生物途径调控的关键点是基于蛋白质-蛋白质的相互作用，而不是基于酶的细胞产物对底物的催化。因此，传统的药物发现方法在它们能够充分解决的目标范围内是有限的。 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分析来实现，这种细胞分析通过计算机视觉的模式和细胞器易位来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反应。此外，生理性的细胞异质性(例如，细胞分裂周期特定的阶段)或表面上随机的细胞异质性可能会压倒常规的高通量高温超导读数。这些都是基于细胞图像的仪器可以极大地增加自动化分析的信息含量的方法的例子。基于细胞图像的筛选的缺点在于，与HTS传统的全孔板阅读器HTS系统每天100,000孔或更多的速率相比，在中等显微镜分辨率(具有10-40倍物镜的大约0.5-2.0微米/米)下成像的仪器通常被限制在每天约25,000个孔。虽然已经报道了更高的放大率，但这些增加通常是通过牺牲分辨率来实现的(例如，甚至更低的放大率目标或大量的相机投放)。 在这里，我们建议将基本图像扫描带宽(以像素/S衡量)比目前的Beckman Coulter IC-100仪器提高10倍，该仪器的原型是由普赖斯博士的学术实验室开发的，首先由Q3 DM Inc.商业化。这一提高将通过应用全新的原理实现，这些原理将自动对焦和图像采集并行化，以扫描长的、不间断的连续运动多色条中的载玻片和微滴定板。这将导致使用中分辨率物镜(10-40倍干式放大倍率)每天筛选超过100,000口井的速度。

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).