CONFOCAL UNIT

该提案的目的是为美国国立卫生研究院支持的弗吉尼亚大学的关键研究项目提供最先进的共聚焦激光扫描显微镜功能。为了实现这一目标，建议购买共聚焦激光扫描显微镜（CLSM），特别是Biorad MRC 1024扫描单元和奥林巴斯IX70倒置显微镜。该仪器将作为一个多用户系统集成到中央核心设施——先进细胞成像设施（ACIF）中，为6名研究人员提供服务，他们在研究中都有迫切的、未满足的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，研究基因产物的定位、功能和相互作用，已经在细胞和发育生物学和遗传学中取得了许多突破性的应用。在这种情况下，参与的教师需要一个高灵敏度的CLSM，能够做时间推移，重复成像的生物材料，最小的光毒性，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）都有需要对多个荧光标记进行三维成像的应用，包括固定制剂的静态成像和活细胞的动态时序成像。这些美国国立卫生研究院支持的研究项目包括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。其他应用还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，现有的仪器存在软件过时、灵敏度不足、无延时功能或仅限于单色成像等问题，无法满足用户的需求。MRC 1024将满足对高灵敏度和快速图像采集时间的仪器的需求，用于在最小光毒性作用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师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 Currently, the available instruments suffer from out of date software, inadequate sensitivity, no time-lapse capability, or are limited to single color imaging and thus are inadequate to meet the users' needs.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.