Cell and Molecular Imaging Core

细胞和分子氧化还原成像核心-项目总结 南卡罗来纳州氧化剂、氧化还原平衡和应激信号(Redox 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)，能够 新方法开发(目标2)和培训(目标3)，以调查人员和中心成员为目标 最先进的基于细胞和组织的显微成像技术。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、活体成像和组织切片的自动成像。 此外，Core还维护和开发了各种分子工具和氧化还原指示剂，供Cobre使用 调查。CMRI配备了以下主要显微镜系统：1)蔡司LSM 880 NLO 配备相干变色龙多光子激光、类星体光谱的多光子/共焦系统 探测和AiryScan超分辨率能力；2)奥林巴斯FluoView FV1200多光子显微镜，具有 光谱物理迈泰深见激光和硅油光学技术用于活体成像；3)奥林巴斯FluoView FV 10i LiV水浸光学活细胞共聚焦显微镜；4)蔡司LSM 510元共聚焦显微镜；5) BD BioSciences Carv II用于视频率成像的圆盘扫描共聚焦显微镜；6)蔡司Axiovert 200m 广场荧光显微镜；7)Perkin-Elmer Vectra Polaris自动定量病理学 成像系统。除了为组织切片定制的Vectra Polaris外，所有的显微镜都是 配备用于温度和气相控制的环境室，以实现非破坏性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- 用于监测离子、电位、自由基产生、吡啶核苷酸还原、 膜通透性、细胞活力(细胞凋亡和坏死)和荧光的亚微米分布 蛋白质和其他荧光记者；2)免疫细胞化学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 微循环、白细胞边际化、线粒体极化和通透性、自由基生成、基因 在活体动物中的表达和其他参数；以及5)高通量、定量多路复用成像 常规和免疫染色的临床和研究标本。标本所需的辅助设备 还提供了准备工作。透射式和扫描式电子显微镜的咨询和服务 和扫描电子显微镜)可通过病理和实验室医学系获得。电脑 工作站提供离线图像处理/分析(ImageJ斐济、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).