Microscopy & Enteroids

项目摘要-核心B-显微镜和肠道(PRASAD和ESTES) 显微镜和肠核的作用是为电子显微镜提供一个集中的设施 项目中所有研究人员的(EM)、荧光显微镜和肠道培养要求 项目。此集成核心具有成本效益，有助于有序、高效地使用资源来完成 及时提出项目建议。此外，核心的使用也保证了一致性和质量 控制所有项目。按照三个项目的常规要求，核心的具体职能将 包括：(1)电子显微镜和荧光显微镜的样品制备，包括荧光、共聚焦和 去卷积显微镜；(2)诺如病毒感染者粪便样本的EM成像 重组颗粒(VLP)、VLP-碳水化合物、VLP-抗体复合体、由感染产生的病毒 培养物，全长p41寡聚体，空的和p41重组的纳米盘；(3)染色和 固定病毒感染细胞的薄片以评估细胞结合和复制；(4)纯度、颗粒的评估 用于低温电磁成像和计算机重建的样品的完整性、浓度和适用性；(5) 使用金标记物的免疫电子显微镜；(6)数据的获取、解释、共享和存档 EM、荧光、共聚焦和去卷积显微镜成像；(7)日程安排和日常EM 维护EM、荧光、共焦和去卷积显微镜；(8)培养和提供 所有项目的媒体和特征良好的人肠道肠样(HIE)培养；(9)准备和 用于所有项目的转基因肠系线的特性；和(10)制备和特性 用于监测感染或宿主细胞过程的表达生物传感器的HIE品系。核心关键人员将与 与计划项目组组长和其他调查人员一起满足显微镜和肠样要求 在所有项目中。总体而言，显微镜和肠腔核心是方案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成果可比性 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.