FLUORESCENCE DIGITAL IMAGING MICROSCOPY & IMAGE

本申请的目的是建立一个中心， 荧光数字成像显微术和图像处理. 华盛顿大学。 主要用户群将使用此 研究荷尔蒙控制基本方面的仪器 细胞内钙和氢的稳态在广泛的范围内， 组织中 受体介导的信号转导研究 内吞作用，细胞生长和分化，趋化性，细胞向 细胞相互作用和细胞-细胞外基质相互作用是 纳入大型主要用户群体提议的研究 每项研究都直接代表一个扩展/新方向 适用于由PHS资助的研究。 因此，应用程序 代表着一个激动人心的机会， 对用户群的研究。 主要研究者 单细胞记录细胞内 Ca 2+和pH值。使用新仪器进行的研究 表示新信息在 细胞钙稳态的激素控制区。 这一领域 包括钙通过质膜进入的机制， 内质网与质膜的相互作用。 以及钙离子进入和流出机制的协调。 的 分析细胞氢代谢，其在趋化性中的作用， 并研究了pHi与（Ca ~（2+））i关系 通过这个提案。 亚细胞分布和调节 （H+）和（Ca ~（2+））i的亚细胞区室是 研究旨在分析受体介导的内囊化，细胞， 细胞外基质相互作用和细胞间通讯。 使用特定的荧光团来研究细胞运输， 这些物质与上述研究直接相关/整合。 所要求的系统设计用于确定三维 荧光团的荧光强度直方图 活细胞的亚细胞区室 干扰光发射。 此外，该系统还提供 能够快速分析显微荧光图像，数字化， 并通过运动的电影和视频图像来分析它们。 因此，该项目代表了一个机会， 用户组开发对理解至关重要的新信息 激素生理学，细胞生物学，以及 肾脏和骨骼疾病。

The purpose of this application is to establish a center for fluorescence digital imaging microscopy and image processing at Washington University. The major user group will employ this instrumentation to study fundamental aspects of hormonal control of cell calcium and hydrogen homeostasis in a broad range of tissues. Studies of signal transduction, receptor mediated endocytosis, cell growth and differentiation, chemotaxis, cell to cell interaction and cell-extracellular matrix interaction are incorporated in the studies proposed by the large major user group Each study represents an extension/new direction directly applicable to research funded by the PHS. Thus, the application represents in exciting opportunity to develop important new thrusts to the research of the user group. The principal investigator has developed technology for single cell recordings of intracellular Ca2+ and pH. The studies proposed using the new instrumentation represent critical extensions required of new information in the area of hormonal control of cell calcium homeostasis. This area includes mechanisms of calcium entry through the plasma membrane, interactions of the endoplasmic reticulum with the plasma membrane. and coordination of calcium entry and efflux mechanisms. The analysis of cell hydrogen metabolism, its role in chemotaxis and the relationship between pHi and (Ca2+)i will also be studied through this proposal. Subcellular distribution and regulation of subcellular compartments of (H+) and (Ca2+)i are major features of studies designed to analyze receptor mediated endocystosis, cell- extracellular matrix interaction and cell to cell communication. The use of specific fluorophores to study cell trafficking of substances are directly related/integrated into the above studies. The system requested is designed to determine the three dimensional histogram of fluorescent intensity of fluorophores associated with subcellular compartments in living cells with correction for interfering light emission. In addition, the system provides the capability to rapidly analyze microfluorimetric images, digitizing, them and analyzing them with cine and video images of movement. Thus, this project represents an opportunity for a large diverse user group to develop new information critical to the understanding of the hormonal physiology, cell biology, and pathophysiology of renal and osseous diseases.