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丰富的知识库转化为新产品以改善人类医学奠定了基础。实现基因组学工作前景的一个关键组成部分是获得充分表征的化合物，这些化合物可以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化学生物学计划提供了有利的基础，使用化合物作为探索生物途径的发现工具，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蛋白质靶点。人们逐渐认识到，生物途径调节的许多关键点都是基于蛋白质-蛋白质相互作用，而不是细胞产物对底物的基于酶的催化。因此，传统的药物发现方法在其能够充分解决的靶标范围方面受到限制。 筛选科学家发现，基于图像的细胞检测可以通过模式和细胞器易位的计算机视觉自动测量蛋白质动态，从而阐明生物途径调节关键点上细胞对化合物的反应。膜和细胞内受体的亚基通常通过重组或易位做出反应。此外，生理性（例如细胞分裂周期特定阶段）或明显随机的细胞异质性可能压倒全孔传统高通量筛选 HTS 读数。这些是基于细胞图像的仪器可以显着增加自动化检测的信息内容的示例。基于细胞图像的筛选的缺点是，中等显微镜分辨率（约 0.5 - 2.0 mu/m，10-40 倍物镜）的仪器成像通常仅限于每天约 25,000 个孔，而 HTS 传统全孔读板机 HTS 系统的成像速度为每天 100,000 个或更多孔。虽然报道了更高的比率，但这些增加通常是通过牺牲分辨率来实现的（例如，甚至更低的放大倍率物镜或大量的相机合并）。 在这里，我们建议将基本图像扫描带宽（以像素/秒为单位）增加 10 倍，比当前的 Beckman Coulter IC-100 仪器增加 10 倍，该仪器的原型是在 Price 博士的学术实验室中开发的，并首先由 Q3DM Inc 进行商业化。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，以扫描长的、不间断的连续运动多色条带中的载玻片和微量滴定板。使用中等分辨率物镜（10-40 倍干放大倍率），这将导致每天超过 100,000 个孔的筛选速度。

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).