Zeiss LSM510 Meta Confocal Microscope

描述(由申请人提供)：这项建议要求一个光谱共焦，蔡司LSM 510与Meta检测器，以促进我们的理解细胞内感染和宿主反应通过使用新的细胞内报告及其应用于NIH优先病原体名单上的感染性病原体。该仪器具有多种荧光定量功能，并可对环境舱内的活体样品数据进行高级分析。该仪器将被安置在专门为此目的而设计的生物安全二级成像中心。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。关联性。新出现的病原体的威胁，以及传染性病原体的潜在武器化，都是我们越来越意识到的问题。虽然在许多病原体的基因组测序方面已经取得了巨大的进步，但基因和功能之间的联系以及它们在感染中的作用仍有待确定。大多数细胞内病原体进入它们的宿主细胞是在宿主起源的膜结合的隔间，即。吞噬菌体。在正常情况下，这种吞噬小体会将其内容物传递给溶酶体，病原体就会被杀死。但细胞内的病原体通过逃逸到胞浆、重塑吞噬小体或阻止其成熟来避免这种情况。这项建议使用新的荧光分析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NIH资助的正在进行的识别新药靶点、疫苗和佐剂的计划。

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.