Xenogen IVIS Imaging System 200

描述(申请人提供)：威斯康星大学麦迪逊分校迫切需要对小动物进行非侵入性实时活体成像。这一需求集中在宿主-微生物相互作用的研究上。进行这些检查所需的设备是Xenogen IVIS成像系统200。这种实时活体成像最大的好处是使用的老鼠要少得多，因为该仪器允许科学家在整个实验过程中跟踪同一只老鼠，而不是在不同的时间点使用和牺牲几只老鼠。当D-荧光素或Coelenterazine被注射并分别被萤火虫或肾虫荧光素酶用作底物时，光从活动物体内发出。光谱成像使用从420 nm到720 nm的不同波长过滤的一系列图像的测量数据，以确定生物发光报告器的深度和位置。IVIS成像系统200包括一个灵敏的CCD相机、一个最大限度地减少入射光的黑暗成像室，以及一个同时允许生物发光和荧光成像的22滤光轮。安装在转塔上的定制镜头提供5级放大，从26厘米到2.9厘米，允许高分辨率(60？m)的详细成像。Living Image专业软件对结果进行分析和量化，甚至可以进行3D漫反射断层成像重建，以可视化内部器官。所有这些功能-带有多个报告器的荧光和生物发光成像、高分辨率放大和3D重建-将为我们的实验提供极大的灵活性。在这群用户之间共享这一特定的仪器是最大限度地利用技术的灵活性的一个很好的例子。这台仪器的成像能力将使用户能够可视化老鼠/微生物的相互作用，以及1)夏威夷鱿鱼Euprymna Scolope和其发光细菌伙伴鱼肝弧菌之间的共生相互作用模型，2)吉普赛蛾中肠的正常微生物区系，3)线虫(Steinenema Carpocapsae)、幼虫阶段昆虫和伽马保护细菌(Xnowhabdus Nematophila)之间的三方相互作用。威斯康星大学麦迪逊分校以在宿主-微生物相互作用领域处于领先地位而闻名；我们的目的是为我们的研究开发这些体内成像模型和试剂，并将它们提供给全国其他研究人员。

DESCRIPTION (provided by applicant): There is a dramatic need for noninvasive real-time in vivo imaging of small animals at the University of Wisconsin-Madison. This need is centered around the study of host- microbe interactions. The equipment required to perform these investigations is the Xenogen IVIS Imaging System 200. The most significant benefit of this real-time in vivo imaging is far fewer mice will be used because the instrument allows a scientist to follow the same mouse throughout an experiment instead of using and sacrificing several mice at various time points. Light is emitted from within a living animal when D-Luciferin or Coelenterazine is injected and used as a substrate by firefly or renilla luciferases, respectively. Spectral imaging uses measurement data from a sequence of images filtered at different wavelengths ranging from 420 nm to 720 nm, to determine the depth and location of the bioluminescent reporter. The IVIS Imaging System 200 includes a sensitive CCD camera, a dark imaging chamber to minimize incident light, and a 22-filter wheel that permits both bioluminescent and fluorescent imaging. Turret-mounted custom lens provide 5 levels of magnification, from 26 cm to 2.9 cm, allowing high-resolution (60 ¿m) detailed imaging. Living Image specialized software analyzes and quantifies the results, and can even perform 3D diffuse tomography reconstructions to visualize internal organs. All of these features - fluorescent and bioluminescent imaging with multiple reporters, high-resolution magnification, and 3D reconstruction - will give us great flexibility with our experiments. Sharing this particular instrument among this group of users is a beautiful example of exploiting the flexibility of a technology to its fullest. This instrument's imaging capabilities will enable users to visualize mouse/microbe interactions as well as the model symbiotic interactions between 1) the Hawaiian squid Euprymna scolopes and its luminous bacterial partner Vibrio fischeri, 2) the normal microbial flora of the gypsy moth midgut, and 3) the tripartite interaction between a nematode (Steinernema carpocapsae), larval stage insects, and a gamma-proteobacterium, Xenorhabdus nematophila. The University of Wisconsin-Madison is well known for leading the field of host-microbe interactions; it is our intent to develop these in vivo imaging models and reagents for our studies, and to make them available to other investigators across the country.