Confocal spinning disk high throughput screening microscope

我们正在申请资金，以获得几个正在进行和计划进行的跨学科研究项目迫切需要的共聚焦成像细胞仪。而且，在过去几年中，高端免疫荧光显微镜的技术要求也在稳步增长，我们目前在这一领域的需求已经无法满足Charité Campus米特现有的显微镜基础设施（柏林三个慈善机构之一）。最近，这导致我们在许多项目中的显微镜实验出现重大延迟，也影响了我们能够生成的数据的质量。此外，由于现有显微镜的技术限制，我们无法建立一些要求更高的实验装置，这将大大有利于现有的项目，并需要建立新的研究方向。最近，显微镜的设计取得了巨大的进步，并出现了许多新的先进应用在生物学中：更灵敏的相机、更大的视野和越来越全自动化的图像采集工作流程，以及新的光学设计，使得从数百个3D样本中长时间采集高质量的共焦图像成为可能。因此，可以在自然环境中随着时间的推移在3D中系统地跟踪单细胞，这对于更好地理解许多与医学直接相关的生物过程至关重要：例子包括癌症的发展，跟踪组织内的免疫反应，而且还涉及细胞生物学中的许多问题，例如需要在类器官中进行实验。成像细胞仪的另一个重要应用将是获得高-高分辨率共聚焦图像的组织学标本从病人，荧光染色与新的多路复用协议，使连续染色相同的幻灯片的几十种蛋白质。用校园现有的设备获取这种图像极其耗时，因此实际上不可行。所包含的图像分析软件包还将允许我们在采集过程中以高通量自动化方式分析数据。总之，共聚焦成像细胞仪将确保尖端显微镜设备的可用性，以实现高通量3D活细胞成像，再加上适用于定量如此大量数据的专用软件，并帮助我们建立新的基于显微镜的分析方法。这也将使参与团体的成员更容易为现有项目获得足够的显微镜时间，依靠校园内的现有设备已经变得越来越困难。在所附的申请表中，我们包括一个详细的计划，如何使用细胞仪，使七个不同的实验室作为共同申请人的许多现有和计划中的研究项目受益。

We are applying for funding to acquire a confocal imaging cytometer that is urgently needed in several ongoing and planned interdisciplinary research projects.The possibilities, but also the technical requirements in high-end immunofluorescence microscopy have steadily expanded during the last few years and our current needs in this area cannot be met anymore with the available microscopy infrastructure on Charité Campus Mitte (one out of 3 Charité sites in Berlin). Recently, this has led to significant delays in our microscopy experiments in many projects and also affected the quality of the data we were able to generate. Furthermore, because of technical limitations of the available microscopes, we were not able to establish some more demanding experimental setups that would have greatly benefitted existing projects and are required to establish new research directions.Recently, enormous progress has been made in the design of microscopes and lots of new advanced applications in biology have emerged: more sensitive cameras, larger field of view and increasingly fully automated image acquisition workflows, as well as new optical designs make it possible to acquire high quality confocal images from hundreds of samples in 3D over long periods of time. Thus, it becomes possible to systematically track single cells in 3D over time in their natural environment, which is essential for a better understanding of many biological processes that are of immediate relevance in medicine: examples include the development of cancer, tracking of the immune response within tissues, but also many questions in cell biology that require for example experiments in organoids.Another important application for the imaging cytometer will be to obtain high-resolution confocal images of histological specimens from patients, fluorescently stained with new multiplexed protocols that enable sequential staining of the same slide for dozens of proteins. Acquisition of such images with the equipment currently available on campus is extremely time-consuming and therefore practically not feasible. The included image analysis package will also allow us to analyze the data on the fly during acquisition in an automated manner in high throughput.In summary, the confocal imaging cytometer will ensure the availability of cutting edge microscopy equipment for high-throughput 3D live-cell imaging coupled with dedicated software appropriate for quantification of such large amounts of data and help help us to establish new microscopy-based assays. It will also make it easier for members of the involved groups to get enough microscopy time for existing projects, which has become increasingly difficult relying on the existing equipment on campus. In the included application forms, we include a detailed plan how the cytometer will be used as benefit many existing and planned research projects of seven different labs that act as coapplicants.