Zeiss LSM510 Meta Confocal Microscope

描述（由申请人提供）：该提案要求使用光谱共聚焦，Zeiss LSM 510（带Meta检测器），通过使用新型细胞内报告基因及其对NIH优先病原体列表中存在的感染因子的应用，来促进我们对细胞内感染和宿主反应的理解。该仪器具有关键功能，可对多种荧光剂进行定量荧光分析，并对环境室中的活样品进行高级数据分析。仪器将存放在专门为此目的设计的专用生物安全2级成像中心。我们有一个由10名联邦政府支持的研究人员组成的小组，他们建议使用这些方法来解决细胞内感染的基本问题。本案无关新出现的病原体的威胁以及传染性病原体的潜在武器化是我们日益认识到的问题。虽然在许多病原体的基因组测序方面取得了巨大进展，但基因和功能之间的联系及其在感染中的作用仍有待确定。大多数细胞内病原体进入宿主细胞的膜结合区室，即。吞噬体在正常情况下，这种吞噬体会将其内容物输送到溶酶体，病原体就会被杀死。但胞内病原体通过逃入胞质溶胶、重塑吞噬体或阻止其成熟来避免这种情况。该建议采用新的荧光检测和新兴的定量荧光显微镜来剖析一系列重要病原体的感染过程。这些数据将扩展正在进行的NIH资助的项目，用于识别新的药物靶点，疫苗和佐剂。

DESCRIPTION (provided by applicant): This proposal requests a spectral confocal, the Zeiss LSM 510 with Meta detector, to advance our understanding of intracellular infection and host response through the use of novel intracellular reporters and their application to infectious agents present on the NIH list of Priority Pathogens. This instrument has key capabilities that allow quantitative fluorescence of multiple fluors and advanced analysis of data from live samples contained in an environmental chamber. The instrument will be housed in a dedicated Biosafety Level 2 Imaging Center designed specifically for this purpose. We have a group of 10 federally-supported investigators who propose to use these methods to address questions fundamental to intracellular infections. Relevance. The threat from emergent pathogens, and the potential weaponization of infectious agents are issues of which we are increasingly aware. While there have been enormous strides in genomic sequencing of many pathogens, the links between gene and function, and their role in infection are still being determined. Most intracellular pathogens enter their host cells in a membrane-bound compartment of host origin, ie. the phagosome. Under normal circumstances this phagosome would deliver its contents to a lysosome and the pathogen would be killed. But intracellular pathogens avoid this by escaping into the cytosol, remodeling the phagosome, or arresting its maturation. This proposal employs novel fluorescence assays and emerging quantitative fluorescence microscopy to dissect out the infection process for a range of important pathogens. These data will extend ongoing, NIH-funded programs for identification of new drug targets, vaccines and adjuvants.