Imaging Resource

用于单分子检测、光学切片的荧光显微镜的最新进展 与物体识别算法结合使用的三维数据采集现在允许 足够的时间和空间分辨率以可视化的顺序招募和细胞内流量 活细胞中的蛋白质、脂类和病原体，包括病毒和细菌。Kirchhausen实验室已经 长期以来，人们对了解毒素、病毒和细菌进入人体的机制很感兴趣 细胞，以及这些过程如何与囊泡小管的组织和细胞内交通有关 膜载体。作为这项工作的一部分，我们已投入大量精力开发成像套件 包含最先进的显微镜和支持软件，适用于数据收集和分析 高空间和时间分辨率。维护100%可访问的映像套件也是当务之急 向所有用户开放，这样经过初步培训后，任何调查员都可以完全灵活地开展工作 独立。 在这份提案中，我们概述了NERCE成像核心设施的计划，该设施将提供访问当代 活细胞和单分子成像的工具和专业知识旨在但不限于定量 细菌和病毒病原体入侵哺乳动物细胞的机制描述 复制，以及与毒素进入细胞有关的分子方面。

Recent developments in fluorescence microscopy geared for single-molecule detection, optical sectioning and three-dimensional data acquisition, used in combination with object-identification algorithms, now allow sufficient temporal and spatial resolution to visualize the sequential recruitment and intracellular traffic of proteins, lipids, and pathogens in living cells including viruses and bacteria. The Kirchhausen laboratory has a longstanding interest in understanding the mechanisms underlying entry of toxins, viruses and bacteria into cells, and how these processes relate to the organization and intracellular traffic of vesiculo-tubular membrane carriers. As part of this effort, we have devoted substantial efforts to developing an imaging suite containing state-of-the-art microscopes and supporting software suited for data collection and analysis with high spatial and temporal resolution. It is also a priority to maintain an imaging suite that is 100% accessible to all users, so that after initial training, any investigator can perform their work with total flexibility and independence. In this proposal we outline plans for a NERCE Imaging Core facility that will provide access to contemporary tools and expertise for live-cell and single molecule imaging aimed towards, but not limited to, quantitative descriptions of mechanisms of invasion for bacterial and viral pathogens into mammalian cells, of viral replication, and of molecular aspects related to toxin entry into cells.