Spinning disk confocal / FRAP microscope for quantitative live cell imaging

用于定量活细胞成像的转盘共焦/FRAP 显微镜

基本信息

批准号：
7792018
负责人：
Torsten Wittmann
金额：
$ 50万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-27 至 2011-05-26
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): We are requesting funds to purchase a state-of-the-art Spinning Disk Confocal Microscope System specifically designed for live cell imaging of mammalian cells and tissues. The instrument will be used by a core group of seven NIH-funded investigators and their laboratories to facilitate fourteen NIH- funded projects. The principle types of experiments for which this microscope system will be used include multidimensional imaging of fluorescently tagged proteins in living cells and tissues, analysis of protein activity by FRET, analysis of protein and organelle dynamics by FRAP or photoactivation, and photoablation or release of caged compounds in living cells or tissues. The system we are requesting includes a Yokogawa CSU-X1 spinning disk scanner on a Nikon Ti-E inverted microscope stand, which has a built-in focus feedback system allowing imaging for many hours at high resolution with no discernable focus drift. The microscope will also be equipped with an environmental chamber, a motorized stage, and CCD cameras for high resolution and high sensitivity imaging. Oil- and water- immersion objectives are requested for the optimal collection of data from live cells or tissues in a 2D or 3D environment at the highest achievable optical resolution. Solid-state laser lines are requested at 440, 488, 514, and 561 nm to permit imaging of all commonly used fluorescent protein variants. In addition, this instrument will be equipped with a pulsed dye laser-based photobleaching / photoablation module extending its usefulness beyond simple observation of live specimens. Currently, no spinning disk microscope system exists at UCSF that has FRAP or photoactivation capabilities, which severely limits the types of experiments that can be performed. It is expected that the major users will utilize the instrument for ~80% time. However, because the instrument will be housed in and managed through the UCSF Parnassus Biological Imaging Development Center, training and use will be available to additional NIH-funded investigators within the UCSF community.

描述（由申请人提供）：我们要求资金购买专门设计用于哺乳动物细胞和组织的活细胞成像的最新旋转磁盘共聚焦显微镜系统。该工具将由由NIH资助的七个研究人员及其实验室组成的核心组来促进14个NIH资助的项目。使用该显微镜系统的主要实验类型包括对活细胞和组织中荧光标记的蛋白质的多维成像，通过FRET分析蛋白质活性，通过FRAP或光活化对蛋白质和细胞器动力学分析，以及在活细胞或组织中蛋白质的化合物的光化或释放。我们要求的系统包括尼生CSU-x1旋转磁盘扫描仪上的尼康Ti-e倒置显微镜支架上，该磁盘倒置的显微镜支架具有内置的重点反馈系统，可在高分辨率上进行数小时的成像，而无需明显的焦点漂移。显微镜还将配备环境室，电动阶段和CCD摄像头，用于高分辨率和高灵敏度成像。要求在2D或3D环境中以最高可实现的光学分辨率在2D或3D环境下从活细胞或组织中收集油性和浸入式目标。在440、488、514和561 nm处请求固态激光系，以允许对所有常用的荧光蛋白质变体进行成像。此外，该仪器将配备基于脉冲染料激光的光漂白 /光化模块，该模块扩展了其有用性，而不是简单地观察活样本。当前，在UCSF上没有旋转磁盘显微镜系统具有FRAP或光激活功能，这严重限制了可以执行的实验类型。预计主要用户将使用该仪器约80％的时间。但是，由于该仪器将通过UCSF Parnassus生物成像开发中心安置并管理，因此将向UCSF社区中的其他NIH资助的调查人员提供培训和使用。