SD COBRE: CELL IMAGING CORE

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 在2006财年，成像核心保持了2005财年所述的共聚焦显微镜利用率的增长。2006财政年度，核心能力的重大进步源于对奥林巴斯FluoView 1000共聚焦显微镜的收购，该显微镜目前位于心血管研究所的设施中。这种新仪器提供了最初包含在COBRE赠款中的FVX显微镜的所有功能。FV 1000通过计算机控制显微镜和共焦采集参数，实现这些功能的自动化。FV 1000允许从三个荧光通道采集，允许评估细胞或组织样品中三种蛋白质的共定位。这三个荧光通道可以被分配给各种各样的细胞染料，其激发波长范围从405 nm（例如核染料DAPI）到675 nm（Cy 5）。这种广泛的染料选择为研究人员提供了利用荧光染料的机会，为他们的研究提供了最高水平的灵敏度和选择性。 FV 1000提供的激发和发射波长的精确定义允许这种灵活性。 核心还安装了一个显微镜安装，生理室，将允许采集共聚焦图像的活细胞受到各种实验条件下，在一系列的时间点。这些实验可以包括被显微注射到活细胞中的荧光底物。酶激活的细胞定位可以通过酶激活后荧光的出现在体内确定。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In FY2006, the Imaging Core maintained the growth in confocal microscopy utilization that was described for FY2005. A major advancement in the capabilities of the Core in FY2006 derived from the acquisition of the Olympus FluoView 1000 confocal microscope which is currently located in the facilities of the Cardiovascular Research Institute. This new instrument offers all the functions of the FVX microscope that was originally included within the COBRE grant. The FV1000 automates these functions with computer control over the microscope and confocal acquisition parameters. The FV1000 allows acquisition from three fluorescence channels, permitting an assessment of the co-localization of three proteins within a cellular or tissue sample. These three fluorescence channels can be assigned to a wide variety of cellular dyes with excitation wavelengths in a range from 405nm (e.g. the nuclear stain DAPI) to 675nm (Cy5). This broad range of dye options offers investigators the opportunity to utilize the fluoro-chrome that provides the highest level of sensitivity and selectivity for their studies. This level of flexibility is permitted by the precise definition of excitation and emission wavelengths that is provided by the FV1000. The Core also installed a microscope mounted, physiologic chamber that will permit the acquisition of confocal images from living cells subjected to a variety of experimental conditions over a series of time points. These experiments can include fluorescent substrates that are micro-injected into viable cells. The cellular localization of enzyme activation can be determined in vivo by the appearance of fluorescence subsequent to enzyme activation.