Zeiss LSM 510 Laser Scanning Confocal Microscope

这项赠款提案寻求资助激光扫描共聚焦显微镜，以使用双重和三重标记抗原对各种蛋白质和细胞进行定位。用户群体的研究兴趣多种多样，从研究胃肠道内的细胞多样性到心脏生理学，但都有共同的兴趣，即测量细胞之间的空间位置，细胞转分化过程中蛋白质表达的变化，以及特定细胞类型中蛋白质的定位。用户群体的研究记录是优秀的。用户组有相当多的经验，并发表了广泛的使用共聚焦显微镜。然而，由于我们目前的显微镜(Biorad MRC600)的局限性、年龄和广泛使用，在当前研究过程中出现的许多形态问题都不能得到充分的解决。这对用户组正在进行的项目是一个严重的限制。需要解决的问题包括检查不同组织中细胞之间的空间关系，细胞器和蛋白质的共同定位以及蛋白质在细胞内的移位。这些问题都需要对表面或细胞内的抗原位置进行双重和三重标记。安装蔡司LSM 510激光共聚焦显微镜是促进严格完成已资助项目的特定目标的理想仪器。蔡司提供多达4个反射光共焦通道配置和1个透射光共焦通道配置(DIC)、关键荧光染料组合的多路跟踪、用于正确三维测量的单独可调针孔、12位数据采集和2048x2048像素分辨率。蔡司为这些研究提供了最好的集成系统。内华达大学的研究在过去的几年里有了戏剧性的增长。蔡司LSM 510的拟议用户群体竞争激烈，代表了内华达大学一些最成功的研究人员。该机构通过每年为服务合同提供支持来认可用户群体的质量。PI在共焦显微镜方面有丰富的经验，将担任内部咨询委员会的主席，并监督仪器的日常运行。该仪器将被安置在现有的由NIH资助的成像核心实验室中。

This grant proposal seeks funding for a laser scanning confocal microscope to perform localization of a variety of proteins and cells using double and triple labeling of antigens. The research interests of the user group are diverse ranging from investigating cellular diversity within the gastrointestinal tract to cardiac physiology, but all have common interests in measuring the spatial apposition between cells, changes in protein expression during cellular transdifferentiation, and the localization of proteins within specific cell types. The research record of the user group is excellent. The user group has considerable experienced and has published extensively using confocal microscopy. However, because of limitations, age and extensive use of our current microscope (Biorad MRC 600) many of the morphological questions that have arisen during the course of current investigations cannot be adequately addressed. This is a serious limitation for the ongoing projects of the user group. Questions that need to be addressed include an examination of the spatial relationships between cells in heterogeneous tissues, the co- localization of organelles and proteins and translocation of proteins within cells. These are questions that all require double and triple labeling of antigen sites on the surface or within cells. The ideal instrument to facilitate rigorous completion of specific aims of already funded projects to incorporate a Zeiss LSM 510 laser confocal microscope. The Zeiss provides up to 4 confocal channel configurations for reflected light plus one for transmitted light (DIC), multi-tracking for critical flurochrome combinations, individual adjustable pinholes for correct 3-dimensional measurements, 12 bit data acquisition and 2048x2048 pixel resolution. The Zeiss offers the greatest integrated system available for these studies. Research at the University of Nevada has grown dramatically over the last number of years. The proposed user group of the Zeiss LSM 510 is highly competitive, representing some of the most successful investigators at the University of Nevada. The institution recognizes the quality of the user group by providing yearly support for a service contract. The PI has had considerable experience with confocal microscopy and will chair the internal advisory committee and oversee the daily operation of the instrument. The instrument will be housed in an existing NIH funded imaging core laboratory.