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Cell and Molecular Imaging Core

细胞和分子成像核心

基本信息

批准号：
10460363
负责人：
John J Lemasters
金额：
$ 19.63万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10460363
关键词：
Address Animals Apoptosis Awareness Bioenergetics Biological Sciences Biology Biomedical Research Cell Membrane Permeability Cell Survival Cell physiology Cells Centers of Research Excellence Clinical Research Communities Computer Workstations Computer software Consultations Custom Data Data Analyses Detection Development Educational workshop Electron Microscopy Ensure Environment Equilibrium Equipment Equipment and supply inventories Event Fluorescence Resonance Energy Transfer Fluorescent Probes Freezing Funding Funding Mechanisms Gases Gene Expression Generations Genetic Grant Histopathology Human Resources Image Image Analysis Imaging technology Immersion International Investigation Ions Laboratories Lasers Leadership Leukocytes Measurement Medicine Mentors Methods Microcirculation Microscope Microscopy Mitochondria Molecular Monitor Necrosis Optics Organism Oxidants Oxidation-Reduction Paraffin Pathology Permeability Phase Preparation Proteins Reporter Research Research Personnel Research Proposals Research Support Resolution Resources Running Scanning Scanning Electron Microscopy Series Services Signal Transduction Silicone Oils Slide Source South Carolina Specimen Stress System Technology Temperature Three-Dimensional Imaging Tissue imaging Tissues Training Training Activity Training and Education United States National Institutes of Health Visualization Water Work base cellular imaging digital imaging electrical potential fluorescence microscope fluorophore high resolution imaging histological image histological slides image processing imaging approach imaging system immunocytochemistry instrument instrumentation intravital imaging intravital microscopy lectures light microscopy liquid crystal polymer live cell imaging member method development microscopic imaging molecular imaging multi-photon multiphoton microscopy multiplexed imaging next generation novel operation optical imaging pathology imaging programs protein distribution pyridine nucleotide quasar second harmonic second harmonic generation imaging skills submicron success targeted imaging tool transmission process

项目摘要

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).

细胞和分子氧化还原成像核心 – 项目摘要南卡罗来纳州 COBRE 的氧化剂、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）、能力新方法开发（目标 2）和培训（目标 3）以研究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。这包括共焦、多光子和超分辨率活体和固定细胞和组织的显微镜检查、活体成像以及组织学载玻片的自动成像。此外，该核心还维护并开发了用于 COBRE 的各种分子工具和氧化还原指示剂调查。 CMRI 拥有以下主要显微镜系统： 1) Zeiss LSM 880 NLO 多光子/共焦系统配备 Coherent Chameleon 多光子激光器、Quasar 光谱检测和Airyscan超分辨率能力； 2) 奥林巴斯 Fluoview FV1200 多光子显微镜用于活体成像的 SpectraPhysics MaiTai DeepSee 激光和硅油光学器件； 3）奥林巴斯Fluoview FV 10i 带水浸光学器件的 LIV 活细胞共焦显微镜； 4）蔡司LSM 510 META共焦显微镜； 5） BD BioSciences CARV II 盘扫描共焦显微镜，用于视频速率成像； 6）蔡司Axiovert 200M 宽视场荧光显微镜； 7) Perkin-Elmer Vectra Polaris 自动定量病理学成像系统。除了专为组织学载玻片定制的 Vectra Polaris 外，所有显微镜均采用配备用于温度和气相控制的环境室，以实现无损 3D 活细胞、组织和生物体的成像。主要应用包括：1）参数的活细胞成像敏感荧光团可监测离子、电位、自由基生成、吡啶核苷酸还原、膜通透性、细胞活力（细胞凋亡和坏死）以及荧光的亚微米分布蛋白质和其他荧光报告基因； 2) 免疫细胞化学组织切片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、白细胞边缘化、线粒体极化和通透性、自由基生成、基因活体动物的表达和其他参数； 5) 高通量、定量多重成像常规和免疫染色的临床和研究标本。标本所需辅助设备还提供了准备工作。透射电子显微镜（TEM）和扫描电子显微镜（TEM）的咨询和服务和 SEM）可通过病理学和检验医学系获得。电脑工作站提供离线图像处理/分析（ImageJ FIJI、Metamorph、IPLab 等软件）。