Microscopy & Enteroids

项目总结-核心B -显微镜检查和肠镜检查（Prasad和Estes） 显微镜和肠样核心的作用是提供一个中央设施，为电子显微镜服务 (EM)、荧光显微镜检查和肠样细胞培养要求 项目这一综合核心具有成本效益，有助于有序和有效地利用资源， 及时提出建议。此外，使用核心允许的一致性和质量 控制所有项目。这三个项目例行要求的核心小组的具体职能将 包括：（1）EM和荧光显微镜的标本制备，包括落射荧光，共聚焦， （2）诺如病毒感染者粪便样本的EM成像，病毒样 重组颗粒（VLP），VLP-碳水化合物，VLP-抗体复合物，由感染的病毒产生的病毒 培养物、全长p41寡聚体以及空的和p41重构的纳米盘;（3）染色的和未染色的纳米盘的EM成像。 病毒感染细胞的固定薄切片，以评估细胞结合和复制;（4）纯度、颗粒 冷冻EM成像和计算机重建样品的完整性、浓度和适用性;（5） 使用金标记的免疫电子显微镜;（6）数据采集，解释，共享和存档 EM，荧光，共聚焦和去卷积显微镜成像;（7）调度和日常EM EM、落射荧光、共聚焦和去卷积显微镜的维护;（8）培养和提供 培养基和特征良好的人小肠类（HIE）培养物;（9）制备和 所有项目的转基因肠细胞系的表征;以及（10）制备和表征 表达生物传感器的HIE细胞系来监测感染或宿主细胞过程。核心关键人员将与 与项目组领导和其他研究人员一起，以满足显微镜和肠镜检查的要求 所有项目。总的来说，显微镜和肠样核心是该计划项目成功的关键组成部分 并将提供关键资源，加强不同组织之间的沟通和结果的可比性， 项目

Project Summary - Core B - Microscopy and Enteroids (Prasad and Estes) The role of the Microscopy and Enteroid Core is to provide a centralized facility to serve the electron microscopy (EM), fluorescence microscopy, and enteroid culture requirements of all of the investigators in the Program Project. This integrated Core is cost-effective and facilitates orderly and efficient use of resources to complete the proposed projects in a timely fashion. In addition, the use of the core allows for consistency and quality control across all projects. The specific functions of the Core, as required routinely by the three projects, will include: (1) Specimen preparation for EM and fluorescent microscopy, including epifluorescence, confocal, and deconvolution microscopy; (2) EM imaging of fecal samples from norovirus-infected persons, virus-like recombinant particles (VLPs), VLP-carbohydrate, VLP-antibody complexes, virus produced from infected cultures, full-length p41 oligomers, and empty and p41-reconstituted nanodiscs; (3) EM imaging of stained and fixed thin sections of virus-infected cells to assess cell binding and replication; (4) Evaluation of purity, particle integrity, and concentration and suitability of samples for cryo-EM imaging and computer reconstructions; (5) Immune electron microscopy using gold labels; (6) Data acquisition, interpretation, sharing, and archiving from the EM, fluorescent, confocal and deconvolution microscope imaging; (7) Scheduling and day-to-day EM maintenance of EM, epifluorescence, confocal and deconvolution microscopes; (8) Cultivation and providing media and well-characterized human intestinal enteroid (HIE) cultures to all Projects; (9) Preparation and characterization of genetically modified enteroid lines for all Projects; and (10) Preparation and characterization of biosensor-expressing HIE lines to monitor infection or host cell processes. Core key personnel will interface with program project group leaders and other investigators to meet the microscopy and enteroid requirements of all Projects. Overall, the microscopy and enteroid core is a key component of success of the program project and will provide critical resources, enhance communication and comparability of results between different projects.