Stereology & Live Cell Imaging Workstation

描述（由申请人提供）：本申请资金用于购买成像工作站，该工作站能够基于待计数特征的多通道荧光识别进行体视学测量、延时活细胞荧光成像和测量青色和黄色荧光蛋白（CFP和YFP）之间的荧光共振能量转移（FRET）。形态测量，如细胞的数量和大小，脑回路中神经元纤维的密度，特定区域或特定细胞上的突触密度，卵泡的数量和阶段是与构成俄勒冈州国家灵长类动物研究中心（ONPRC）多个NIH资助的赠款重点的条件相关的生理变化的重要决定因素。如果不遵循现代体视学的采样和分析技术，基于二维显微图像估计这些参数是非常费力的，并且受到偏差的影响。因此，体视学兼容的成像系统是必要的研究我们的调查。对于其他项目，使用绿色荧光蛋白和它们的突变体已经为鉴定感兴趣的基因的细胞内位置、易位和功能创造了机会。一个特别有吸引力的应用是可视化和测量之间的相互作用 生物分子在生理相关的状态，在真实的时间，使用FRET，特别是CFP和YFP之间。这是不可能的与我们目前可用的仪器，但我们的NIH资助的研究人员必须。该工作站将成为位于俄勒冈州健康与科学大学西校区的ONPRC成像和形态学核心的一部分，并将使该核心能够提供现在不可能提供的服务，或者在准确性和速度方面将得到显着提高。一个内部咨询委员会将制定程序并监督成像工作站的一般运作。主要研究者将负责新成像工作站的安装、培训、调度、使用和维护。该机构致力于维护仪器，并提供监督、支持工作人员工资和培训。

DESCRIPTION (provided by applicant): This application requests funds for purchasing an imaging workstation that enables stereological measurements based on multichannel fluorescence identification of features to be counted, time-lapse live cell fluorescence imaging and measuring Fluorescence Resonance Energy Transfer (FRET) between cyan and yellow fluorescent proteins (CFP and YFP). Morphometric measurements, such as numbers and sizes of cells, density of neuronal fibers in brain circuits, density of synapses in specific areas or on specific cells, number and stages of ovarian follicles are important determinants of physiological changes associated with conditions that constitute the focus of research of multiple NIH-funded grants at the Oregon National Primate Research Center (ONPRC). Estimating these parameters based on two-dimensional microscopic images is very laborious and is affected by biases, if sampling and analysis techniques of modern stereology are not followed. A stereology compatible imaging system is therefore necessary for the research of our investigators. For other projects, use of green fluorescent proteins and their mutants has created opportunities for identifying the intracellular location, translocation and function of genes of interest. A particularly attractive application is the visualization and measurement of the interaction between biomolecules in physiologically relevant states, in real time, using FRET, especially between CFP and YFP. This is not possible with the instruments currently available to us, but necessary for our NIH-funded investigators. This workstation will be part of the Imaging and Morphology Core of ONPRC located on the West campus of the Oregon Health & Sciences University and will enable the core to provide services that are either not possible now, or that will be significantly improved in accuracy and speed. An internal advisory committee will establish procedures and oversee the general operation of the imaging workstation. The principal investigator will be in charge of the installation, training, scheduling, use and maintenance of the new imaging workstation. The institution is committed to maintenance of instrument and providing oversight, support for staff salary and training.