Microscopy and Flow Cytometry Core

显微镜和流式细胞术核心

基本信息

批准号：
9978922
负责人：
Chang-UK Lim
金额：
$ 22.35万
依托单位：
UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9978922
关键词：
3-Dimensional Affect Air Anatomy Animal Model Animal Organ Animals Antibodies Apoptosis Applications Grants Biological Carbon Dioxide Cell Aging Cell Count Cell Cycle Progression Cell Death Cell Separation Cells Centers of Research Excellence Chemicals Color Computer software Core Facility Cryoultramicrotomy Data Analyses Diagnosis Equipment Flow Cytometry Fluorescence Fluorescence Resonance Energy Transfer Genetic Histopathology Human Human Pathology Human Resources Image Image Analysis Immunofluorescence Immunologic Immunohistochemistry In Vitro Ions Laboratories Lasers Lesion Light Manuscripts Measurement Metabolism Microscope Microscopic Microscopy Modification Molecular Target Morphology Paraffin Pathologic Pathologist Pathology Pathology Report Pharmaceutical Preparations Phase Preparation Protocols documentation Reagent Research Design Research Personnel Research Project Grants Resources Running Scanning Services Sorting - Cell Movement South Carolina Specimen Handling Speed Stains Sterility Techniques Temperature Time Tissue Sample Tissues Training Universities Video Microscopy animal tissue cell growth chemical genetics design experience experimental study flexibility fluorescence imaging fluorescence microscope human tissue in vivo instrument instrumentation live cell imaging medical schools microsystems programs ratiometric reconstruction small molecule success targeted treatment tumor vibration

项目摘要

ABSTRACT The Microscopy and Flow Cytometry Core (MFCC) of the COBRE Center for Targeted Therapeutics (CTT) provides services to all four CTT research projects and other investigators supported by the CTT and other IDeA programs at the University of South Carolina (USC). These services include training in microscopy and flow cytometry, advice on study design, with an emphasis on the analysis of cellular drug effects, and assistance with the instrumentation usage, both at the Core and at other USC facilities (in particular at the USC School of Medicine), as well as expert assistance in anatomic pathology of human and animal tissues. The available equipment includes: Carl Zeiss LSM700 Laser scanning confocal microscope with 4 lasers functioning from 405 to 633 nm, equipped for live cell imaging (with environmental chamber), standard point scanning, 3D reconstruction, FRAP, FRET, time lapse and tile/stitching; a Leica Microsystems AS MDW fluorescence microscope optimized for live cell imaging with temperature and CO2 control; two Leica Microsystems DM IRE2 fluorescent microscopes, which are motorized for z-stack acquisition and one of which has an environmental chamber. Olympus IX81 fluorescence microscope with workstation running Metamorph software is used for fluorescence imaging and deconvolution of wide field fluorescence images; it also has a Sutter Instruments Smartshutter and rapid filter changing wheels for ratiometric measurement of intracellular ion changes. There are 3 Carl Zeiss Axiovert 200 microscopes and an Olympus upright BX41 microscope, each with a color camera. All microscopes are in dedicated rooms and on vibration isolation tables. Flow cytometric equipment includes the BD FACS ARIA III for cell sorting, BD LSR II Flow Cytometer with high-throughput option and a 96-well plate loader, and Beckman Coulter FC500 flow cytometer. Users may operate the microscopes and one of the flow cytometers independently, after training by the co-directors. Many standard protocols have been established for immunohistochemistry and immunofluorescence analysis of cells and tissues, and for the analysis of cell growth, cell death and cell cycle progression. The core also provides services of an experienced anatomic pathologist, with a broad experience in gross pathology and histopathology in human and animal studies. The pathology core services include advice on study design and specimen processing including paraffin and cryosectioning, routine H&E staining, histochemical stains, and immunohistochemical analysis. Pathology reports will include macroscopic and microscopic findings, diagnosis of lesions, discussion of significant findings, and biological significance of the pathological effects of genetic modifications and experimental compounds in different animal organs, as well as tumor morphology in animal models and in human tissue samples. The core personnel offer advice on the choice of instruments and experimental protocols, provide training and assist with preparation of manuscripts and grant proposals.

抽象的 COBRE 靶向治疗中心 (CTT) 的显微镜和流式细胞术核心 (MFCC) 为 CTT 和其他 IDeA 支持的所有四个 CTT 研究项目和其他研究人员提供服务南卡罗来纳大学 (USC) 的课程。这些服务包括显微镜和流程培训细胞计数、研究设计建议，重点是细胞药物效应分析以及协助仪器使用情况，无论是在核心还是在其他南加州大学设施（特别是在南加州大学医学），以及人类和动物组织解剖病理学方面的专家协助。可用的设备包括： Carl Zeiss LSM700 激光扫描共焦显微镜，配备 4 个激光器，405 至 633 nm，配备活细胞成像（带环境室）、标准点扫描、3D 重建、FRAP、FRET、延时和平铺/拼接； Leica Microsystems AS MDW 荧光针对活细胞成像优化的显微镜，具有温度和 CO2 控制；两台 Leica Microsystems DM IRE2 荧光显微镜，用于 z 堆栈采集，其中之一具有环境室。带有运行 Metamorph 软件的工作站的 Olympus IX81 荧光显微镜用于宽视场荧光图像的荧光成像和反卷积；它还有一个萨特仪器智能快门和快速滤光片更换轮，用于细胞内离子变化的比例测量。那里包括 3 个 Carl Zeiss Axiovert 200 显微镜和一个 Olympus 正置 BX41 显微镜，每个显微镜都配有彩色摄像头。所有显微镜都位于专用房间和隔振台上。流式细胞仪设备包括用于细胞分选的 BD FACS ARIA III、具有高通量选项的 BD LSR II 流式细胞仪和 96 孔板装载机和贝克曼库尔特 FC500 流式细胞仪。用户可以操作显微镜和流程之一经过联合主任的培训后，可以独立使用细胞仪。已经建立了许多标准协议细胞和组织的免疫组织化学和免疫荧光分析，以及细胞生长分析，细胞死亡和细胞周期进展。该核心还提供经验丰富的解剖病理学家的服务，在人类和动物研究的大体病理学和组织病理学方面拥有丰富的经验。病理核心服务包括研究设计和标本处理（包括石蜡和冷冻切片）、常规 H&E 染色、组织化学染色和免疫组织化学分析。病理报告将包括宏观和微观发现、病变诊断、重要发现的讨论以及生物学基因修饰和实验化合物对不同动物的病理影响的意义器官，以及动物模型和人体组织样本中的肿瘤形态。核心人员报价关于仪器和实验方案选择的建议，提供培训并协助准备手稿和资助提案。