Two-photon microscope for intravital imaging

描述（由申请人提供）：由致病细菌和病毒引起的传染病对全球健康构成严重挑战。每年约有290万人死于艾滋病，约200万人死于肺结核，200万人死于细菌引起的腹泻。新出现的传染性病原体，如西尼罗河病毒和竞技场病毒，随着它们传播到地球仪以前未受影响的地区，构成了新的威胁。可能出现的快速传播的H5 N1禽流感毒株可能导致毁灭性的流行病。耶鲁大学和东北部其他卓越中心的科学家致力于了解这些疾病并开发新疗法。体外宿主-病原体相互作用的研究提高了对这些疾病的分子认识.尽管宿主-病原体相互作用越来越清楚，但对这些疾病如何在体内传播的潜在机制知之甚少。在缺乏体内环境的情况下，体外获得的结果的相关性往往是有限的。在这里，我们建议建立一个国家的最先进的双光子成像核心设施下的生物安全三级条件（BSL 3）的可视化宿主-病原体在活生物体的相互作用。活体成像将允许可视化感染的传播和真实的发生的免疫反应。包括人源化小鼠在内的小动物模型可以忠实地重建这些疾病的人类发病机制。新的荧光标记技术使得可视化包括病毒在内的单个病原体并监测其在活生物体中的时空传播成为可能。生物体中感染性病原体和应答宿主免疫细胞的成像有望彻底改变我们对疾病发病机制的理解，并允许合理设计新的治疗方法。

DESCRIPTION (provided by applicant): Infectious diseases caused by pathogenic bacteria and viruses pose a serious challenge to global health. Each year ~2.9 million people die from AIDS, ~2 million from tuberculosis and 2 million from bacterially caused diarrhea. Emerging infectious pathogens like West Nile and Arena viruses pose new threats as they spread into previously unaffected areas around the globe. The possible emergence of a rapidly spreading avian flu strain H5N1 could lead to a devastating epidemic. Scientists at Yale University and other centers of excellence in the northeast are committed to the understanding of these diseases and the development of novel therapies. The study of the host- pathogen interactions in vitro has improved the molecular understanding of these diseases. Despite the increasing clarity in host-pathogen interactions, very little is known about the underlying mechanisms of how these diseases spread in vivo. The relevance of results gained in vitro in the absence of the vivo context is often limited. Here we propose the establishment of a state of the art two-photon imaging core facility housed under biosafety level three conditions (BSL3) for the visualization of host-pathogen interactions in living organisms. Intravital imaging will allow the visualization of the spreading of the infection and the immune responses as it happens in real time. Small animal models including the humanized mouse allow a faithful reconstruction of the human pathogenesis of these diseases. Newly available fluorescent labeling techniques make it possible to visualize individual pathogens including viruses and monitor their temporal and spatial spreading in the living organism. The imaging of infectious pathogens and the responding host immune cells in living organism is expected to revolutionize our understanding of disease pathogenesis and allow the rational design of novel therapies.