Live Cell Confocal Microscope for FRAP/PA

描述（由申请人提供）：本提案的目标是获得一台配备FRAP和光激活的旋转盘共聚焦显微镜，用于测量活细胞中的动态过程。达特茅斯医学院和学院的生物化学和生物科学系主要和次要使用该仪器。他们的研究项目旨在了解基本细胞过程的分子机制，如染色体分离、肌动蛋白组织、核转运和膜运输。圆满完成这些用户的nih资助项目的既定目标需要在高空间和时间分辨率和最小光毒性的活细胞中检查动态细胞过程。本方案中描述的自旋盘共聚焦显微镜是一种最先进的仪器，通过使用成熟的技术（横河自旋盘共聚焦头；尼康倒置显微镜；光子仪器光漂白/激活激光；即兴软件），以最小的光暴露，以快速的速度（高时间分辨率）提供实时共聚焦（高空间分辨率），满足这些需求。它将是同类仪器中唯一一个服务于整个达特茅斯生命科学界的仪器。公共卫生相关性：本提案的目标是深入了解各种人类疾病状态的分子机制，如肿瘤细胞非整倍体、转移和与健康问题（如胰岛素在糖尿病中的作用）相关的膜生物学。显微镜的独特属性赋予了研究人员观察活细胞动态过程的无与伦比的能力，并且可以通过理解这些基本细胞机制的动态来深入了解疾病病因学。

DESCRIPTION (provided by applicant): The goal of this proposal is to acquire a spinning-disc confocal microscope equipped for FRAP and photoactivation to be used for the measurement of dynamic processes in live cells. The major and minor users of this instrument are in the Departments of Biochemistry and Biological Sciences at Dartmouth Medical School and College. Their research projects are aimed at understanding molecular mechanisms of fundamental cellular processes such as chromosome segregation, actin organization, nuclear transport, and membrane trafficking. Satisfactory completion of the stated goals of the NIH-sponsored projects of these users requires examination of dynamic cellular processes in live cells with high spatial and temporal resolution and minimal phototoxicity. The spinning-disc confocal microscope described in this proposal is a state of the art instrument that satisfies these needs by using proven technology (Yokogawa spinning-disc confocal head; Nikon inverted microscope; Photonic Instruments photobleaching/activating laser; Improvision software) to provide real-time confocality (high spatial resolution) at rapid rates (high temporal resolution) with minimal light exposure. It would represent the only instrument of its kind to serve the entire Dartmouth life science community. PUBLIC HEALTH RELEVANCE: The goal of this proposal is to gain insight into the molecular mechanisms that underlie various human disease states such as tumor cell aneuploidy, metastasis, and membrane biology related to health issues such as insulin action in diabetes. The unique attributes of the microscope requested give the investigators unparalleled ability to observe dynamic processes in living cells, and insight into disease etiology can emerge from understanding the dynamics of these fundamental cell mechanisms.