Cell & Molecular Imaging Core

细胞和分子成像核心D -John Lemasters，MD，PhD，核心负责人 摘要 细胞和分子成像核心D为共聚焦、多光子和明场显微镜提供主要支持， 眼镜蛇调查员。成像核心包含以下主要显微镜系统：1）蔡司LSM 880 NLO共焦/多光子显微镜配备有具有类星体的相干变色龙多光子激光器 用于光谱分析的光电倍增管阵列; 2）Olympus FV 1200多光子显微镜，其具有Spectra- 物理MaiTai多光子激光器和硅油光学器件用于活体成像，3）Olympus FV 10 i LIV活细胞 4）Zeiss LSM 510 Meta激光扫描共聚焦显微镜， 显微镜;和5）BD CARV II圆盘扫描共聚焦显微镜，用于视频速率成像，以及 传统的宽视野显微镜。所有显微镜都配备了环境室， 温度和气相控制以允许对活体进行高分辨率非破坏性三维成像 细胞、组织和有机体。该仪器的主要应用包括：1）活细胞成像， 参数敏感的荧光团，用于监测离子、电势、氧和氮自由基的产生， 吡啶核苷酸还原，线粒体和质膜通透性，细胞活力 （凋亡和坏死），荧光蛋白标记和其他参数; 2）活体显微镜监测 微循环，白细胞集落，线粒体极化和通透性，自由基生成，基因 表达和其他参数; 3）免疫染色的荧光和明场成像 组织样品; 4）荧光共振能量转移（FRET）以表征和量化相互作用 5）Duolink邻近连接测定，以研究内源性分子之间的相互作用; 表达的蛋白质。辅助设备包括一个数字暗室设施，用于数字化，分析和标签 图像;三维和四维图像分析和体绘制的软件和计算机工作站;组织 用于标本制备的培养罩和培养箱;以及用于 在多孔板上生长的培养细胞中进行平行测量。成像核心服务将促进 各个COBRE项目的成功，并将为初级调查人员提供培训和帮助 研究与COBRE整体主题相关的氧化应激和应激信号。在第一阶段， COBRE，CMI Core D为COBRE 44篇出版物中的29篇和35份赠款申请中的26篇做出了贡献 investigators.

Cell & Molecular Imaging Core D —John Lemasters, MD, PhD, Core Leader Abstract Cell & Molecular Imaging Core D provides major support in confocal, multiphoton and brightfield microscopy to COBRE investigators. The Imaging Core houses the following major microscope systems: 1) a Zeiss LSM 880 NLO confocal/multiphoton microscope equipped with a Coherent Chameleon multiphoton laser with Quasar photomultiplier array for spectral analysis; 2) an Olympus FV1200 multiphoton microscope with Spectra- Physics MaiTai multiphoton laser and silicone oil optics for intravital imaging, 3) an Olympus FV10i LIV live cell confocal microscope with water immersion optics, 4) a Zeiss LSM 510 META laser scanning confocal microscope; and 5) a BD CARV II disk-scanning confocal microscope for video-rate imaging as well as conventional widefield microscopy. All microscopes are equipped with environmental chambers for temperature and gas phase control to allow high resolution non-destructive 3-dimensional imaging of living cells, tissues and organisms. Major applications of this instrumentation include 1) live cell imaging of parameter-sensitive fluorophores to monitor ions, electrical potentials, oxygen and nitrogen radical generation, pyridine nucleotide reduction, mitochondrial and plasmalemmal membrane permeability, cell viability (apoptosis and necrosis), fluorescent protein labeling and other parameters; 2) intravital microscopy to monitor microcirculation, leukocyte margination, mitochondrial polarization and permeability, radical generation, gene expression and other parameters in living animals; 3) fluorescence and brightfield imaging of immuno-stained tissue samples; 4) fluorescence resonance energy transfer (FRET) to characterize and quantify interactions between specific molecules; 5) Duolink proximity ligation assay to study interactions between endogenously expressed proteins. Ancillary equipment includes a digital darkroom facility for digitizing, analyzing and labeling images; software and computer workstations for 3- and 4-D image analysis and volume rendering; tissue culture hoods and incubators for specimen preparation; and fluorescence and absorbance plate readers for parallel measurements in cultured cells grown on multi-well plates. Imaging core services will promote the success of the individual COBRE projects and will also provide training and assistance to junior investigators studying oxidative stress and stress signaling related to the overall theme of this COBRE. In Phase 1 of the COBRE, CMI Core D contributed to 29 of 44 publications and 26 of 35 grant applications by COBRE investigators.