LASER SCANNING CONFOCAL IMAGING SYSTEMS

本提案中要求的资金将用于建立一个激光 扫描共聚焦显微镜设施（LSCM） 罗得岛医院中心研究实验室。 尼康PCM 2000年系统将大大扩展一个主要的研究能力， 由六名NIH资助的研究人员组成的用户组， 尖端的仪器和技术，将用于 使用一系列对比模式研究活的和固定的细胞和组织 和荧光探针。 各种各样的组织和分离/培养 将分析细胞类型，包括肝、脑、脾、肠 粘膜和网状内皮系统的细胞（以及其他）。 的 尼康PCM 2000系统包括：（a）尼康Eclipse E 800研究 正置显微镜和尼康TE-200倒置显微镜 （配置明场相位收缩、差分干涉 对比度和荧光显微镜光学），（B）多波长 通过光纤耦合的激发能力，氩气 离子/HeNe激光照明源;（c）多波长检测 通过高分辨率双通道荧光检测器系统;（d） 包括硬盘存档能力;（e）通过以太网传输数据 邻近和/或远程预先存在的图像分析工作站;以及 (f)三维重建软件。 提出的具体申请 这种仪器可能包括：（a）检测和鉴别 荧光探针在单标记、双标记或三标记样品中的浓度;（B） 用重要或pH敏感的细胞内活性监测 荧光染料或与荧光标签缀合的离子;和（c）光学 用于三维的嵌入式试样的厚切片 细胞和组织成分的重建。 尼康PCM 2000系统 将满足不同生物医学群体的迫切需要， 科学家的研究计划将大大受益于 这是非常需要的仪器。

Funds requested in this proposal will be used to establish a Laser Scanning Confocal Microscope facility (LSCM) within the Division of Central Research Laboratories at Rhode Island Hospital. The Nikon PCM 2000 System will significantly extend the research capabilities of a major user group comprising of six NIH-funded investigators by providing cutting-edge instrumentation and technologies which will be utilized to study living and fixed cells and tissues using a range of contrast modes and fluorescent probes. A diverse array of tissues and isolated/cultured cell types will be analyzed including liver, brain, spleen, intestinal mucosa, and cells of the reticuloendothelial system (among others). The Nikon PCM 2000 System incorporates: (a) the Nikon Eclipse E 800 Research Upright Microscope and the Research Nikon TE-200 Inverted Microscope (configured with bright field phase contract, differential interference contrast, and fluorescence microscopy optics), (b) multiple wave length excitation capability by means of a fiber optically coupled, Argon Ion/HeNe laser illumination source; (c) multiple wave length detection through a high resolution, two channel fluorescence detector system; (d) including hard disk archiving capability; (e) data transfer via Ethernet to adjacent and/or remote pre-existing image analysis workstations; and (f) three dimensional reconstruction software. Specific applications made possible by this instrumentation include: (a) detection and discrimination of fluorescent probes in single-, double-, or triple-labeled samples; (b) monitoring of intracellular activities with vital or pH sensitive fluorescent dyes or ions conjugated to fluorescent tags; and (c) optical sectioning of thick sections of embedded specimens for three dimensional reconstruction of cellular and tissue elements. The Nikon PCM 2000 System will address the immediate needs of a diverse group of biomedical scientists whose research programs will benefit significantly by the availability of this much needed instrumentation.