Primary Cell and Tissue Processing and Imaging Core

原代细胞和组织处理和成像核心

• 批准号: 8097279
• 负责人: Pawel J Kolodziejski
• 金额: $ 15.14万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 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/8097279
• 关键词: Cell Culture Techniques; Cell Line; Cells; Charge; Chimeric Proteins; Computer-Assisted Image Analysis; Confocal Microscopy; Culture Techniques; Detection; Electron Microscopy; Ensure; Epithelial Cells; Equipment and supply inventories; Floor; Fluorescence; Fluorescence Microscopy; Fluorescent Antibody Technique; Histology; Human; Human Resources; Image; Image Analysis; Immunohistochemistry; Label; Laboratories; Lasers; Life; Light; Microscopic; Microscopy; Molecular; Photobleaching; Primary Cell Cultures; Process; Proteins; Recovery; Research Personnel; Running; Sampling; Services; Structure of parenchyma of lung; Techniques; Thick; Three-Dimensional Image; Tissues; Transmission Electron Microscopy; cellular imaging; cryogenics; image reconstruction; monolayer; research study; tissue processing

The purpose of Primary Cell, Tissue Processing and Imaging Core (Core C) is to provide centralized and expert handling and processing of primary cells, tissues, and to serve as an imaging facility. This Core will be in charge of establishing and maintaining techniques for culturing primary human cells, including airway epithelial cells, and will provide molecular cellular, histological and ultrastructural information for each of the experimental projects. It is highly beneficial to the Center that these activities be performed in a centralized fashion, as it would be difficult and inefficient to run these highly specialized functions by separate laboratories. The Specific Aims of Core C are: Aim 1: The Core will provide expert, centralized initial processing and characterization of human samples. It will transform these samples into primary cells in culture. The Core will develop a cryogenic bank of primary cells, ensure appropriate storage, inventory, and coordinate their distribution to Center investigators. Aim 2: The Core will provide molecular, cellular, histological and ultrastructural information for each of the experimental projects. Core C will perform primary cell culturing, histology, immunohistochemistry, fluorescence microscopy deconvolusion microscopy, live cell imaging, confocal microscopy and electron microscopy. Additional services, including three-dimensional image analysis of fluorescently labeled cells using deconvolution microscopy, will also be available. The facilities available for the Core are all located within close proximity of the investigators' laboratories on the 5th and 6th floor of Baylor's main building. The Core has fluorescence and light microscopy with computer-assisted image analysis and deconvolution microscopy with three-dimensional image analysis capability. In addition, physically located within the Center is the Seymour Lieberman Electron Microscopy Laboratory, which is a dedicated facility for transmission electron microscopy and morphometric analysis. Specialized techniques will be used to localize specific proteins in primary cells, cell lines, and lung tissues. For monolayer cell culture analysis, light microscopic level using immunohistochemistry and, in cases where fluorescent antibody detection is used or when fluorescent fusion proteins are utilized, deconvolution microscopy will be employed in conjunction with computerassisted three-dimensional image reconstruction and analysis. Confocal microscopy will be used for thick tissue sections, for some live cell imaging experiments and for some of the experiments requiring laser, e.g., fluorescence recovery following photobleaching. In addition, Core personnel will carry out initial tissue processing and analysis.

原代细胞、组织处理和成像核心（核心C）的目的是提供集中和 专业处理和加工原代细胞、组织，并用作成像设备。这一核心将 负责建立和维护培养原代人体细胞的技术，包括气道 上皮细胞，并将提供分子细胞，组织学和超微结构的信息，为每一个 实验项目。这些活动以集中的方式进行，对中心非常有益。 时尚，因为将这些高度专业化的功能分开运行是困难和低效的。 laboratories.核心C的具体目标是：目标1：核心将提供专家，集中的初始 人类样本的处理和表征。它将把这些样本转化为原代细胞， 文化核心将建立一个原代细胞的低温库，确保适当的储存、库存， 协调他们分发给中心调查员。目标2：核心将提供分子，细胞， 每个实验项目的组织学和超微结构信息。 核心C将进行原代细胞培养、组织学、免疫组织化学、荧光显微镜检查 去卷积显微术、活细胞成像、共聚焦显微术和电子显微术。额外 服务，包括使用去卷积对荧光标记细胞进行三维图像分析 显微镜，也将提供。可供核心区使用的设施都位于 贝勒主楼第五和第六层的调查员实验室。核心 荧光和光学显微镜与计算机辅助图像分析和去卷积显微镜 具有三维图像分析能力。此外，位于中心内的是 西摩·利伯曼电子显微镜实验室，这是一个专门的设施，为透射电子 显微镜和形态测定分析。专门的技术将用于定位特定的蛋白质， 原代细胞、细胞系和肺组织。对于单层细胞培养分析，使用光学显微镜水平 免疫组织化学，并且在使用荧光抗体检测的情况下或当荧光抗体检测时， 利用融合蛋白，反卷积显微镜将与计算机辅助 三维图像重建和分析。共焦显微镜将用于厚的 组织切片，对于一些活细胞成像实验和一些需要激光的实验， 例如，在一个实施例中，光漂白后的荧光恢复。此外，核心人员将进行初始组织 处理和分析。