A system for automated high-content screening and laser manipulation of single cells.

荧光显微镜是一种重要的技术，被广泛用于研究无数的细胞过程。许多荧光探针已被设计用于可视化和分析大范围的分子，从离子到核酸和蛋白质。这些探针通常用于细胞中，以报告固定细胞和活细胞以及组织和整个生物体中分子的存在和定位。因此，荧光显微镜是了解不同分子和蛋白质之间复杂的细胞内相互作用的基本工具，只需分析这些工程探针发出的光即可。*申请人目前每天将该技术用于广泛的应用，其中使用单个图像来表征数十或数百个细胞的行为。然而，一些关键的研究要求细胞的数量要高得多（例如，在许多不同的条件下平行测试几种药物的效果）。为此，我们需要一个机动系统，可以在大型细胞培养室中长时间成像数百万个细胞。该功能要求显微镜能够在获得图像之前自动重新聚焦样品。然而，大多数系统通过处理图像，通过算法分析照片的不同特征来实现这一点，但这种方法无法提供我们所需的准确性。我们正在申请的系统使用内部激光，该激光在细胞基底上反射，并提供关于样品位置的精确信息以保持焦点。定位和重新聚焦的自动化允许在大量细胞和条件下以高分辨率询问细胞行为的精细细节。* 最后，我们需要使用外部激光单独操纵细胞。例如，我们可以搜索表现出预定义表型的细胞，并仅照亮这些细胞。我们要求的系统配置允许集成/添加我们已经到位的外部激光系统。这些能够适应和扩展新技术发展的外部端口是我们迫切需要的独特功能，无法作为交钥匙解决方案获得。这一基础设施还将允许培训高素质的人才，开发创新的工程解决方案，以解决生物问题。

Fluorescence microscopy is an essential technology that is widely used to study a myriad of cellular processes. Numerous fluorescent probes have been designed to visualize and analyze a large range of molecules, from ions to nucleic acids and proteins. These probes are commonly used in cells to report the presence and localization of molecules in both fixed and live cells, as well as tissue and whole organisms. Fluorescence microscopes, thus, are fundamental tools to understand the intricate intracellular interactions between different molecules and proteins, by simply analysing light emitted by these engineered probes.******The applicants currently use this technology for a wide range of applications on a daily basis, where individual images are used to characterize the behaviour of tens or hundreds of cells. Nevertheless, several critical studies require the number of cells to be much higher (e.g. testing the effects of several drugs in parallel in many different conditions). For this, we are requesting a motorized system that can image millions of cells for extended periods of time in large cell culture chambers. This feature requires the microscope to be able to automatically refocus the sample before images are obtained. However, most systems do this by processing images, by analysing different features of the photos with algorithms, but this approach does not provide the accuracy we require. The system we are applying for uses an internal laser that is reflected on the cell substrate and provides precise information about the position of the sample to maintain focus. The automation of positioning and refocusing allows interrogation of fine details of cell behaviour with high resolution in a large number of cells and conditions. ******Finally, we need to use external lasers to manipulate cells individually. For example, we may search for cells that exhibit a predefined phenotype and illuminate only those cells. The configuration of the system that we request allows integration/addition of external laser systems that we already have in place. These external ports that enable adaptation and expansion for novel technology developments are unique features for which we have a pressing need and cannot be obtained as a turn-key solution. This infrastructure will also allow training of highly qualified personnel in the development of innovative engineering solutions to solve biological questions.********