Cell and Molecular Imaging Core

细胞和分子氧化还原成像核心-项目摘要 南卡罗来纳州COBRE在氧化剂，氧化还原平衡和应激信号（氧化还原COBRE）细胞和 分子成像氧化还原核心（CMRI）提供先进的技术和专业知识（目标1）， 新方法开发（目标2）和培训（目标3），针对研究者和中心成员， 最先进的细胞和组织显微成像技术。这包括共焦、多光子和超分辨率 活的和固定的细胞和组织的显微镜检查、活体成像和组织学载玻片的自动成像。 此外，核心维护和开发各种分子工具和用于COBRE的氧化还原指示剂 调查事务所CMRI包含以下主要显微镜系统：1）Zeiss LSM 880 NLO 多光子/共焦系统，配备相干变色龙多光子激光器，类星体光谱 2）Olympus Fluoview FV 1200多光子显微镜， SpectraPhysics MaiTai DeepSee激光和硅油光学器件用于活体成像; 3）Olympus Fluoview FV 10 i LIV活细胞共聚焦显微镜，带水浸光学系统; 4）Zeiss LSM 510 Meta共聚焦显微镜; 5） BD BioSciences CARV II盘式扫描共聚焦显微镜，用于视频速率成像; 6）Zeiss Axiovert 200 M 宽视野荧光显微镜;和7）Perkin-Elmer Vectra Polaris自动定量病理学 成像系统。除了为组织学切片定制的Vectra Polaris外，所有显微镜均 配备温度和气相控制环境室，可实现无损3D 活细胞、组织和生物体的成像。主要应用包括：1）参数的活细胞成像- 灵敏的荧光团，用于监测离子、电势、自由基生成、吡啶核苷酸还原， 细胞膜通透性、细胞活力（凋亡和坏死）以及荧光标记的亚微米分布。 蛋白质和其它荧光报告物; 2）用于免疫细胞化学的组织切片的高分辨率成像 和荧光蛋白分布; 3）荧光共振能量转移（FRET）和DuoLink， 表征和量化特定分子之间的相互作用; 4）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;和5）高通量，定量多路成像， 常规和免疫染色的临床和研究标本。标本所需的辅助设备 还提供了制备。透射和扫描电子显微镜（TEM）的咨询和服务 和SEM）可通过病理学和实验室医学系获得。计算机 工作站提供离线图像处理/分析（ImageJ FIJI、Metamorph、IPLab和其他软件）。

Cell & Molecular Redox Imaging Core – Project Summary The South Carolina COBRE in Oxidants, Redox Balance and Stress Signaling (Redox COBRE) Cell and Molecular Imaging Redox Core (CMRI) provides advanced technologies and expertise (Aim 1), capabilities for new method development (Aim 2), and training (Aim 3) to target investigators and Center members for state-of- the-art cell- and tissue-based microscopic imaging. This includes confocal, multiphoton and super-resolution microscopy of live and fixed cells and tissues, intravital imaging, and automated imaging of histological slides. Additionally, the Core maintains and develops a variety of molecular tools and redox indicators for use in COBRE investigations. The CMRI houses the following major microscope systems: 1) Zeiss LSM 880 NLO multiphoton/confocal system equipped with a Coherent Chameleon multi-photon laser, Quasar spectral detection and Airyscan super-resolution capability; 2) Olympus Fluoview FV1200 multiphoton microscope with SpectraPhysics MaiTai DeepSee laser and silicone oil optics for intravital imaging; 3) Olympus Fluoview FV 10i LIV live cell confocal microscope with water immersion optics; 4) Zeiss LSM 510 META confocal microscope; 5) BD BioSciences CARV II disk-scanning confocal microscope for video-rate imaging; 6) Zeiss Axiovert 200M wide-field fluorescence microscope; and 7) Perkin-Elmer Vectra Polaris Automated Quantitative Pathology Imaging System. Except for the Vectra Polaris, which is customized for histological slides, all microscopes are equipped with environmental chambers for temperature and gas phase control to allow non-destructive 3D imaging of living cells, tissues and organisms. Major applications include: 1) live cell imaging of parameter- sensitive fluorophores to monitor ions, electrical potentials, radical generation, pyridine nucleotide reduction, membrane permeability, cell viability (apoptosis and necrosis), and the submicron distribution of fluorescent proteins and other fluorescent reporters; 2) high resolution imaging of tissue sections for immunocytochemistry and fluorescent protein distribution; 3) fluorescence resonance energy transfer (FRET) and DuoLink to characterize and quantify interactions between specific molecules; 4) intravital microscopy to monitor microcirculation, leukocyte margination, mitochondrial polarization and permeability, radical generation, gene expression and other parameters in living animals; and 5) high throughput, quantitative multiplexed imaging of conventionally and immunostained clinical and research specimens. Ancillary equipment required for specimen preparation is also provided. Consultation and services for transmission and scanning electron microscopy (TEM and SEM, respectively) are available through the Department of Pathology & Laboratory Medicine. Computer workstations provide offline image processing/analysis (ImageJ FIJI, Metamorph, IPLab and other software).