Histology Core

组织学核心

• 批准号: 8306847
• 负责人: Clifford A Lowell
• 金额: $ 19.22万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 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/8306847
• 关键词: Animals; Architecture; Autoimmune Diseases; Cell Count; Cell physiology; Cells; Data; Detection; Disease model; Freezing; Histologic; Histology; Human Resources; Image; Immune; Immunology; Inflammation; Inflammatory; Location; Lymphoid; Methods; Microscope; Microscopy; Nucleic Acids; Paraffin; Paraffin Embedding; Regulation; Running; Sampling; Scanning; Serum; Signaling Molecule; Staining method; Stains; System; Technology; Tissue Embedding; Tissue Fixation; Tissues; adaptive immunity; animal resource; base; cryostat; cytokine; digital; experience; fluorescence microscope; mouse model; tissue preparation

Each of the projects of this PPG application utilize mouse models to assess innate immune cell function in infectious, inflammatory and autoimmune disease. As such, each project will rely on histologic analysis of tissues from experimental animals to determine innate immune cell numbers, location, and function (such as cytokine secretion). Histologic analysis of lymphoid architectures in relationship to the disease models being studied will also be a key method for each of the projects. Since the PPG envisions extensive sharing of animal resources, ideas and technologies, it is important than standardized methods of tissue preparation, immune cell staining and immunofluorescent detection be utilized by all four projects in the PPG. This will allow rational comparison of results between the projects. The Histology Core will be run through Dr. Lowell's group (Project #3) and will perform the following analyses: 1) standard tissue fixation, paraffin embedding, sectioning and histochemical staining; 2) frozen tissue embedding sectioning and immunohistochemical staining; 3) frozen tissue embedding with immunofluorescent staining and fluorescent cell localization; 4) Multiplex cytokine analysis on serum, tissue and cell supernatant samples using Luminex technologies. Core personnel have extensive experience with all of the above methods, as described in the Preliminary data. The core is equipped with an OTF5000 cryostat for frozen tissue sectioning and a standard Leica microtomb for paraffin sections. Routine microscopy is done with an Eclipse TS100F epi-fluorescent inverted microscope with Cool-Scan 3.0 mega pixel digital imagining system. Access to larger upright Zeiss epi-fluorescent microscopes is available through the UCSF Immunology Imaging Core. This PPG core will also contain a staining station for routine histologic stains and will maintain a battery of mAbs for immunostaining. Multiplex bead based analysis of both cytokines, intracellular signaling molecules and nucleic acids can be performed on the new BioPlex200 analyzer that will be available to PPG personnel.

该PPG应用程序的每个项目都使用鼠标模型来评估先天免疫细胞 在感染性，炎症性和自身免疫性疾病中起作用。因此，每个项目都将依靠 实验动物的组织学分析以确定先天免疫细胞数量， 位置和功能（例如细胞因子分泌）。淋巴结构的组织学分析 与正在研究的疾病模型的关系也将是每个项目的关键方法。 由于PPG设想了动物资源，思想和技术的广泛共享，因此很重要 比标准化的组织制备方法，免疫细胞染色和免疫荧光检测 PPG中的所有四个项目都可以使用。这将允许合理比较结果 项目。 组织学核心将通过洛厄尔博士的小组（项目＃3）运行，并将执行 经过分析：1）标准组织固定，石蜡嵌入，切片和组织化学 染色； 2）嵌入切片和免疫组织化学染色的冷冻组织； 3）冷冻组织 嵌入免疫荧光染色和荧光细胞定位； 4）多重细胞因子 使用Luminex Technologies分析血清，组织和细胞上清液样品。 核心人员在上述所有方法上都有丰富的经验，如 初步数据。该核心配备了OTF5000低温恒温器，用于冷冻组织切片和A 石蜡切片的标准Leica Microtomb。常规显微镜使用Eclipse TS100F完成 带有酷扫描3.0 Mega Pixel数字想象系统的Epi荧光倒倒显微镜。访问 较大的直立Zeiss Epi荧光显微镜可通过UCSF免疫学成像获得 核。该PPG核心还将包含一个用于常规组织学污渍的染色站，并保持 电池电池可进行免疫染色。基于多重珠的两种细胞因子，细胞内的分析 可以在新的Bioplex200分析仪上进行信号分子和核酸 可用于PPG人员。