CONFOCAL UNIT

这项提议的目标是为弗吉尼亚大学正在进行的NIH支持的关键研究项目提供最先进的激光扫描显微镜能力。为了实现这一目标，建议购买一台共焦激光扫描显微镜(CLSM)，特别是带有奥林巴斯IX70倒置显微镜的Biorad MRC1024扫描单元。该仪器将被整合到中央核心设施--高级细胞成像设备(ACIF)，作为一个多用户系统，为6名研究人员提供服务，他们在研究中都有迫切的、未得到满足的CLSM需求。利用绿色荧光蛋白(GFP)及其颜色变体作为融合蛋白，以及其他荧光标记来研究基因产物的定位、功能和相互作用，在细胞和发育生物学和遗传学方面已经产生了许多开创性的应用。在这种情况下，参与的教员需要一种高灵敏度的激光共聚焦显微镜，能够以最小的光毒性对生物材料进行延时、重复成像，并能够成像几种不同颜色的荧光标记。6个主要用户(Adler、Barroso、Corwin、Green、Hays和Keller)都有需要以3-D方式成像多个荧光标记的应用，既包括固定制剂的静态成像，也包括活细胞的动态时间序列成像。这些由美国国立卫生研究院资助的研究项目涉及对细胞结构和功能的几个方面至关重要的分子的位置、运动和相互作用进行成像。其他应用包括对发育生物学中形态发生和细胞运动的重要方面进行成像。目前，现有的仪器软件陈旧，灵敏度不足，没有延时能力，或仅限于单色成像，不能满足用户的需求。MRC1024将满足对具有高灵敏度和快速图像采集时间的仪器的需求，以便在光毒作用最小的情况下对活细胞进行近实时和延时成像。这些能力对于我们的教员在研究和研究培训中保持竞争力至关重要。

The objective of this proposal is to provide state of art confocal laser scanning microscopy capabilities for key, ongoing NIH-supported research programs at the University of Virginia. To accomplish this goal, purchase of a confocal laser scanning microscope (CLSM), specifically a Biorad MRC 1024 scanning unit with an Olympus IX70 inverted microscope, is proposed. This instrument will be integrated into a central core facility, the Advanced Cellular Imaging Facility (ACIF), as a multi-user system, serving 6 investigators, all of whom have pressing, unmet needs for CLSM in their research. Use of green fluorescent protein (GFP) and its color variants as fusion proteins, as well as other fluorescent markers, to study the localization, function, and interaction of gene products, has resulted in many ground-breaking applications in cell and developmental biology and genetics. In this context, the participating faculty need a CLSM of high sensitivity, capable of doing time- lapse, repeated imaging of living material with minimal photoxicity, and capable of imaging several fluorescent markers of different colors. The 6 major users (Adler, Barroso, Corwin, Green, Hays, and Keller) all have applications requiring imaging multiple fluorescent labels in 3-D, both static imaging of fixed preparations and dynamic time-series imaging of living cells. These NIH-supported research projects involve imaging the locations, movements, and interactions of molecules critical in several aspects of cell structure and function. Other applications involve imaging important aspects of morphogenesis and cell movement in developmental biology. Currently, the available instruments suffer from out of date software, inadequate sensitivity, no time-lapse capability, or are limited to single color imaging and thus are inadequate to meet the users' needs. The MRC 1024 will fill the need for an instrument with high sensitivity and fast image acquisition times for near real time and time-lapse imaging of living cells with minimal phototoxic effects. These capabilities are essential for our faculty to remain competitive in research and in research training.