MULTIUSER CONFOCAL MICROSCOPE FACILITY

我们正在请求设备建立多用户共焦 显微镜设备。本申请书中建议的每一个项目 需要使用共焦显微镜，但目前还没有提供，网址为 科罗拉多大学医学院。借阅试行研究 设备已经直接证明了大多数 建议的项目。 拟议设施有两个基本组成部分：a 激光扫描共聚焦显微镜与宽带共聚焦 现有显微镜的附件。每种类型的显微镜都有一定的 与其他类型相比的优势；这两种类型都需要执行 建议的项目。LSCM将设在一个专门的中心 该空间将通过耗资24,000美元的机构进行全面翻新 承诺。宽带共焦附着体及成像工作站 将是可移动的，因此可以安装在用户自己的实验室中。 这些设备最初将由4个用户中的9个共享 医学院的不同部门。行政规定 将鼓励增加新用户。每个用户目前都有资金 至少一项小灵通研究拨款；小灵通研究的总数 目前发放给所有参与调查人员的补助金为13笔。 为了最大限度地利用共焦显微镜，我们 将采集的图像传输到其他工作站(已可用) 用于处理和分析，如有必要。这一安排将继续 共焦计算机免费进行图像采集，避免了 图像处理和分析造成的瓶颈。 大多数项目涉及相对分子的荧光显微镜。 厚实的准备；既有固定的(分段和整体)，也有活的 将使用材料。共焦显微镜的主要优势是 它能够排除从平面上方和下方发射的光子 聚焦，从而允许解析否则将 被背景照明模糊。这种光学切片允许 从多焦点获得的图像进行三维重建 飞机。 总而言之，科学项目、管理结构和 机构承诺将提供灵活、节省时间和成本的 有效地解决了实现研究目标的问题 多个用户。

We are requesting equipment to establish a multiuser confocal microscope facility. Each of the projects proposed in this application requires use of a confocal microscope, but none is currently available at the University of Colorado Medical School. Pilot studies with borrowed equipment have demonstrated directly the feasibility of most of the proposed projects. There are two essential components to the proposed facility: a laser scanning confocal microscope (LSCM) and a broadband confocal attachment for existing microscopes. Each type of microscope has certain advantages compared to the other; both types will be necessary to perform the proposed projects. The LSCM will be located in a dedicated central space which will be fully renovated through a $24,000 institutional commitment. The broadband confocal attachment and imaging workstation will be mobile, and so can be housed in a user's own laboratory. The devices will initially be shared by nine users from four different departments of the Medical School. Administrative provisions will encourage the addition of new users. Every user is currently funded by at least one PHS research grant; the total number of PHS research grants currently awarded to all participating investigators is 13. To maximize the availability of the confocal microscopes, we will transfer acquired images to other workstations (already available) for processing and analysis, if necessary. This arrangement will keep the confocal computers free for image acquisition, and avoid the bottleneck created by image processing and analysis. Most projects involve fluorescence microscopy of relatively thick preparations; both fixed (sectioned and whole mount) and living materials will be used. The key advantage of the confocal microscope is its ability to exclude photons emitted from above and below the plane of focus, thereby permitting resolution of structures which would otherwise be blurred by background illumination. This optical sectioning allows three-dimensional reconstructions from images obtained in multiple focal planes. In summary, the scientific projects, administrative structure, and institutional commitments will provide a flexible, time saving, and cost effective solution to the problem of meeting the research goals of multiple users.