Advanced Imaging Core

高级成像核心-摘要 高级成像核心的总体目标是为DDRCC调查人员提供先进的 成功的高端细胞和组织成像和分析所需的技术、方法和培训， 包括共焦、多光子和超分辨率显微镜、活体成像和自动成像 组织学切片。具体目标是：1)为DDRCC调查人员提供最先进的光学 显微镜；2)开发新的、一线的成像应用，以满足正在出现的科学需求 DDRCC成员；以及3)提供专家指导、教育、培训和咨询 光学成像技术使DDRCC调查人员能够进行科学发现，以及更大 MUSC研究社区。核心包含以下主要显微镜系统：1)蔡司LSM 880 NLO 配备类星体光谱探测和快速空气扫描超分辨率的多光子/共焦系统 能力；2)奥林巴斯FluoView FV1200多光子显微镜，采用硅油光学技术进行活体成像；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 Meta 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 200米宽视场荧光显微镜；7)Perkin-Elmer Vectra Polaris自动化 定量病理成像系统。除了为组织学切片定制的Vectra Polaris外，所有 显微镜配备了用于温度和气相控制的环境室，以允许非 对活细胞、组织和生物体进行破坏性的3D成像。主要应用包括1)活细胞超级 指示参数的荧光团的分辨率成像以监测离子、电势、自由基的产生 吡啶核苷酸还原、膜通透性、细胞存活率(凋亡和坏死)和亚微米 荧光蛋白和其他荧光记者的分布；2)组织切片的高分辨率成像 用于免疫细胞化学和荧光蛋白分布；3)荧光共振能量转移(FRET) 和DuoLink来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，白细胞边集，线粒体极化和通透性，自由基产生， 活体动物的基因表达和其他参数；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。计算机工作站提供离线 图像处理/分析(Bitstream Imaris、ImageJ斐济、Metamorph、IPLab等)。亲身实践和 当前和下一代成像方法的教学培训通过研讨会、演示和 查尔斯顿生物科学光学显微镜研讨会，为期两年，为期一周。先进的成像核心 这些服务促进了DDRCC研究人员对消化系统和肝脏疾病的研究取得成功。

Advanced Imaging Core - Abstract The overall goal of the Advanced Imaging Core is to provide DDRCC investigators with sophisticated technologies, methods and training required for successful, high-end cell and tissue-based imaging and analysis, including confocal, multiphoton and super-resolution microscopy, intravital imaging, and automated imaging of histological slides. Specific Aims are: 1) Provide DDRCC investigators with state-of-the-art capabilities in optical microscopy; 2) Develop novel, front-line imaging applications that address the emerging scientific needs of DDRCC members; and 3) Provide expert mentoring, education, training, and consultation on sophisticated optical imaging technologies that enable scientific discovery by DDRCC investigators, as well as the greater MUSC research community. The Core houses the following major microscope systems: 1) Zeiss LSM 880 NLO multiphoton/confocal system equipped with Quasar spectral detection and Fast Airyscan super-resolution capability; 2) Olympus Fluoview FV1200 multiphoton microscope with 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 widefield fluorescence microscope; and 7) Perkin-Elmer Vectra Polaris Automated Quantitative Pathology Imaging System. Except for 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 super- resolution imaging of parameter-indicating 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 immuno-stained clinical and research specimens. Computer workstations provide offline image processing/analysis (Bitstream Imaris, ImageJ FIJI, Metamorph, IPLab and others). Hands-on and didactic training in current and next-generation imaging approaches are provided by seminars, demos and a biennial 1-week Charleston Workshop on Light Microscopy for the Biosciences. The Advanced Imaging Core services promote the success of DDRCC investigator research on digestive and liver disease.