SD COBRE: CELL IMAGING CORE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 2006财年，成像核心保持了2005财年描述的共焦显微镜使用率的增长。2006财年核心能力的一个重大进步来自于收购奥林巴斯FluoView 1000共焦显微镜，该显微镜目前位于心血管研究所的设施中。这台新仪器提供了FVX显微镜的所有功能，这些功能最初包括在Cobre赠款中。FV1000通过计算机控制显微镜和共焦采集参数实现了这些功能的自动化。FV1000允许从三个荧光通道获取，从而可以评估细胞或组织样本中三种蛋白质的共存情况。这三个荧光通道可以分配给激发波长从405 nm(例如核染色DAPI)到675 nm(Cy5)的各种细胞染料。这种广泛的染料选择为研究人员提供了利用氟铬的机会，为他们的研究提供了最高水平的灵敏度和选择性。FV1000提供的激发和发射波长的精确定义允许这种程度的灵活性。Core还安装了一个安装在显微镜上的生理室，它将允许在一系列时间点上采集处于各种实验条件下的活细胞的共焦图像。这些实验可以包括显微注射到活细胞中的荧光底物。通过酶激活后荧光的出现，可以在体内确定酶激活的细胞定位。

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.