Shared Resource: Microscopy, Imaging and Cytometry Resources

摘要 显微镜、成像和细胞学资源(MICR)核心的目的是增强同级 审查KCI成员的资助研究活动，其研究需要共聚焦显微镜，FLOW 细胞学、小动物成像及相关技术。为此，在本供资期间， CORE为癌症中心成员发表的92份同行评议出版物提供了便利。MICR核心是分组的 在基础研究核心群中，除了MICR核心之外，还包括蛋白质组学和 动物模型和治疗评估核心。能力和服务包括：共聚焦显微镜、 多光子显微镜、常规光学显微镜、FRET和FRAP、TIRF和原子力显微镜， 活体小动物成像(包括SPECT、CT、PET、X射线和光学成像)、流式细胞术和 比率计量学分析(例如，细胞内pH和离子测量研究)，以及3-和4- 三维图像重建和定量测量。磁共振成像也可以通过 MICR和MR中心之间的合作努力。我们的先进成像系统由蔡司LSM组成 410共焦显微镜，配备六条激光线路，PerkinElmer UltraView ERS旋转盘共焦 配备三条激光线路的显微镜，蔡司LSM-510Meta NLO共焦显微镜八条激光 线和多光子成像，徕卡TCS-SP5 MP配备了九条激光线和多光子能力， 新获得的奥林巴斯/布鲁克TIRF/AFM定制显微镜和尖端的蔡司LSM 780共焦 显微镜有六条常规激光线路和Intune激光器，可调谐的白光激光器超过200 激光测线。我们的流式细胞仪系统包括Becton Dickinson(BD)LSR II和BD FACSCanto II Flow 细胞计、BD FACS Vantage SE SORP细胞分选仪和新收购的Amennis ImageStreamX Mark II 成像流式细胞仪。该设施还通过蔡司提供传统的荧光显微镜 最先进的蔡司活细胞成像显微镜--Axiophot三摄像机光学显微镜 Apotome模块能够进行四维成像(时间上为3D)。体内小动物成像单元是一种 通过在Vivo中添加Bruker而向核心提供的新升级技术组合 Extreme和MS FX Pro小型动物光学/X射线成像仪、西门子Inveon SPECT/CT和协和 MicroPET R4小型动物扫描仪。如上所述，小动物核磁共振也可以通过 MICR和位于隔壁的MR中心之间的合作努力。MICR的分析系统 核心包括八个Apple和PC工作站，配备各种分析软件和照片质量 打印机和MIP，分子成像门户。MIP是基于UNIX的中央服务器，存储容量为45 TB 容量和74 TB自动磁带备份系统，以及托管 高级图像分析软件MatLab™和Definiens™，两个许可证，每个许可证最多允许两个用户 同时访问每个软件。

ABSTRACT The purpose of the Microscopy, Imaging and Cytometry Resources (MICR) Core is to enhance the peer reviewed funded research activities of KCI members whose research requires confocal microscopy, flow cytometry, small animal imaging and related techniques. To this end, during the current funding period the Core has facilitated 92 peer-reviewed publications by the cancer center members. The MICR Core is grouped in the Basic Research Core Cluster which, in addition to the MICR Core, includes the Proteomics and the Animal Model and Therapeutics Evaluation Cores. Capabilities and services include: confocal microscopy, multiphoton microscopy, conventional light microscopy, FRET and FRAP, TIRF and Atomic Force Microscopy, in vivo small animal imaging (including SPECT, CT, PET, x-ray, and optical imaging), flow cytometry and ratiometric analyses (e.g., intracellular pH and ion measurement studies), as well as three- and four- dimensional image reconstruction and quantitative measurements. MR imaging is also available through a collaborative effort between MICR and the MR Center. Our advanced imaging systems consist of a Zeiss LSM 410 confocal microscope equipped with six laser lines, a PerkinElmer UltraView ERS spinning disk confocal microscope equipped with three laser lines, a Zeiss LSM-510 META NLO confocal microscope with eight laser lines and multiphoton imaging, a Leica TCS-SP5 MP equipped with nine laser lines and multiphoton capability, a newly acquired Olympus/Bruker TIRF/AFM custom microscope and the cutting-edge Zeiss LSM 780 confocal microscope with six conventional laser lines and the InTune laser, a tunable white light laser with over 200 laser lines. Our flow cytometry systems include Becton Dickinson (BD) LSR II and BD FACSCanto II flow cytometers, a BD FACS Vantage SE SORP cell sorter and the newly acquired Amnis ImageStreamX Mark II imaging flow cytometer. The facility also provides conventional fluorescent microscopy through a Zeiss Axiophot Triple-Camera Photomicroscope, a Zeiss live cell imaging microscope with the state-of-the-art Apotome module capable of four dimensional imaging (3D in time). The in vivo small animal imaging unit is a newly upgraded combination of technologies made available to the core through the addition of Bruker In Vivo Extreme and MS FX Pro small animal optical/x-ray imagers, Siemens Inveon SPECT/CT and Concord MicroPET R4 small animal scanners. As mentioned above, small animal MRI is also available through collaborative efforts between MICR and the MR Center located next door. The analytical systems of the MICR Core include eight Apple and PC workstations with various analytical software and photographic quality printers and the MIP, Molecular Imaging Portal. MIP is a UNIX based central server with 45 TB storage capacity and a 74 TB robotic tape backup system as well as a dedicated applications server hosting the advance image analysis software, Matlab™ and Definiens™, with two licenses each allowing up to two users access to each software simultaneously.