Infectious Diseases Research Technologies Core - RML

传染病研究技术核心 - RML

• 批准号: 9566797
• 负责人: Robert Hohman
• 金额: $ 611.86万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 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/9566797
• 关键词: Address; Antigens; Arts; Bacteria; Bacterial Genome; Bioinformatics; Biological Preservation; Biomedical Technology; Biotechnology; Cells; Chemicals; Collaborations; Communication; Communities; Complex; Computers; Confocal Microscopy; Congresses; Contracts; Core Facility; Data; Data Analyses; Data Collection; Databases; Development; Dideoxy Chain Termination DNA Sequencing; Dimensions; Discipline; Educational Background; Educational workshop; Electron Microscopy; Electrons; Environment; Equipment; Experimental Designs; Explosion; Flow Cytometry; Freeze Substitution; Freezing; Gene Expression Profiling; Genomics; Human; Human Resources; Image; Image Analysis; Imagery; Imaging technology; Immune; Immunology; Industry; Infectious Diseases Research; Institutes; Internet; Internships; Intramural Research; Intramural Research Program; Investigation; Knowledge; Label; Laboratories; Lasers; Lead; Light; Link; Macromolecular Complexes; Medical; Methods; Microscope; Microscopy; Mission; Molecular Biology; Molecular Structure; Morphologic artifacts; National Institute of Allergy and Infectious Disease; Optics; Phase; Photography; Preparation; Proteomics; Protocols documentation; Research; Research Personnel; Research Support; Reservations; Resolution; Rest; Running; Sampling; Scheme; Scientist; Services; Site; Sorting - Cell Movement; Specimen; Structure; Surveys; Techniques; Technology; Testing; Three-dimensional analysis; Time; Training; Training and Education; Transportation; United States National Institutes of Health; Vendor; Viral; Virus; Visual; Work; animation; cryogenics; data management; design; detector; digital; electron tomography; exome; experience; experimental study; high throughput analysis; improved; instrumentation; interest; light microscopy; meetings; microwave electromagnetic radiation; new technology; next generation sequencing; outreach; pathogen; pathogen genome; programs; protocol development; satisfaction; skills; structural biology; success; tomography; tool; web app; web site

In 2017, this project provides state-of-the-art research technologies that are developed, validated, and then applied in support of NIAID research. Technologies developed outside the NIH are likewise tested, evaluated, validated and, if appropriate, incorporated into the technology portfolio of the intramural program. Electron Microscopy The Electron Microscopy (EM) Unit provides expertise in advanced electron microscopy (EM)-related techniques and technologies to support the ultra-structural imaging needs of the DIR scientists both in Bethesda and at the Rocky Mountain Laboratories (RML). Real-time viewing of samples by video image sharing is available to all NIAID users, enabling scientists in Bethesda and Rockville to view samples simultaneously with the RML microscopy staff. The facility provides sample preparation, imaging, and analysis, ranging from basic structural studies and immune-localization of selected antigens to high resolution and three-dimensional analyses for a wide array of specimens. Optimal preservation of specimens is one of the major challenges in the field of EM, and the EM Unit uses a variety of methods, protocols, and equipment to accommodate different preparative and imaging needs. Conventional techniques for sample preparation inherently introduce structural changes in specimens, which can lead to artifacts. The EM Unit has put much effort into this critical first step as evidenced by the development of application-specific chemical and cryogenic preparative techniques to optimally preserve structures of interest for visualization and interpretation. These techniques in combination with new advanced imaging technologies and the ability to plunge freeze for near native state preservation, now allows visualization of macromolecular complexes, viruses, bacteria, and host-pathogen interactions at high resolution and in three dimensions. Concurrently, our bioinformatics component continues to evolve for high-resolution analyses of increasingly complex structures, and more efficient and comprehensive data collection for higher and better throughput. The broad educational background of the EM Unit staff, along with the opportunity to participate in microscopy meetings and hands-on workshops have been critical for expanding the repertoire of techniques. The Unit has expertise for both tomographic data collection and high resolution analysis tools, protocol development especially for immunology, optimizing parameters for techniques including data collection schemes, microwave-assisted freeze substitution (MWFS), and correlative light and electron microscopy (CLEM). Collaborations with various experts in structural biology and cryo-electron tomography (Cryo-ET) continue to provide both analytical assistance vital for determining viral and macromolecular structures, and automated tomographic alignments for improving database efficiency and workflow. New personnel include interns and contract hires who bring both microscopy and data management expertise. On-going developments in both techniques (i.e. immune-labeling, serial section SEM, montage tomography) and technologies (i.e. direct electron detector and the Volta phase plate) have expanded capabilities with significant improvement in 3D visualization and resolution potential at 3 angstroms. Collaborations resulting in improved data analysis, data management, specimen preparation and data collection have been invaluable for efficient developing highly specialized skills. Flow Cytometry Project-specific research applications are developed for flow cytometry analysis and sorting in BSL-2 and BSL-3 environments. RML Genomics Unit The RML Genomics Unit provides DIR investigators with expertise in experimental design, sample preparation, experimentation, and data analysis for high throughput genomic technologies such as Next Generation Sequencing (NGS), and qPCR. The RML Genomics Unit (GU) specializes in gene expression analysis of both host and/or pathogen, de novo assembly of bacterial genomes, and SNP/INDEL analysis of pathogen genomes and human exomes. The Unit also performs Sanger sequencing in a high throughput manner and assists investigators to troubleshoot difficult sequencing regions. The Units focus is to help the investigator optimize their use of genomic technologies. By using knowledge of the technologies and experience in experimental design and data analysis, the Unit can determine the ideal number of samples and replicates, the best technological platform, and the appropriate analysis to achieve the desired results in the most efficient manner. Visual and Medical Arts The Visual and Medical Arts Unit (VMA) at Rocky Mountain Laboratories (RML) provides highly specialized and advanced technologies to support the Division of Intramural Researchs (DIR) research program. The unit offers a wide variety of digital technologies, involving a comprehensive suite of illustration, design services, photography, 3D animation, video, and new media to document scientific findings. In addition, the VMA provided scientific communication to the Office of the Director, NIAID for presentation to scientists, the public as well as US lawmakers. The Visual and Medical Arts (VMA) Unit works in close collaboration with the research staff to provide the highest quality illustrations and images to assist in the visual communication of complex scientific ideas and findings. In addition to supporting research in the DIR, the VMA also provides varied media support for the National Institute of Allergy and Infectious Diseases (NIAID) Office of the Director. Some of these tasks have included creating visuals used in testimony to congress and foreign dignitaries.

在2017年，该项目提供了最先进的研究技术，这些技术经过开发，验证，然后应用于支持NIAID研究。同样对NIH之外开发的技术进行了测试，评估，验证，并在适当的情况下纳入壁内计划的技术组合中。 电子显微镜 电子显微镜（EM）单元提供了高级电子显微镜（EM）相关技术和技术的专业知识，以支持贝塞斯达和落基山实验室（RML）的DIR科学家的超结构成像需求。所有NIAID用户都可以通过视频图像共享对样品进行实时查看，这使贝塞斯达和罗克维尔的科学家能够与RML显微镜工作人员同时查看样品。 该设施提供样品制备，成像和分析，范围从基本的结构研究和选定抗原的免疫定位到高分辨率和三维标本的三维分析。标本的最佳保存是EM领域的主要挑战之一，EM单元使用多种方法，协议和设备来满足不同的制备和成像需求。样品制备的常规技术固有地引入了样品的结构变化，这可能导致伪影。 EM单元已经在这一关键的第一步中付出了很多努力，这是通过开发特定的化学和低温制备技术来证明的，以最佳地保留感兴趣的可视化和解释结构。 这些技术与新的先进成像技术结合使用，并能够冻结近距离状态保存的能力，现在可以在高分辨率和三个维度下可视化大分子复合物，病毒，细菌和宿主 - 病原体相互作用。 同时，我们的生物信息学组成部分继续发展，以对日益复杂的结构进行高分辨率分析，并更有效，更全面的数据收集，以获得更高，更好的吞吐量。 EM单位员工的广泛教育背景以及参加显微镜会议和动手讲习班的机会对于扩大技术曲目至关重要。 该单元具有用于层析成像数据收集和高分辨率分析工具的专业知识，协议开发，特别是用于免疫学，优化了包括数据收集方案，微波炉辅助冻结替代（MWFS）以及相关光和电子显微镜（Clem）的技术。与各种结构生物学和冷冻电子层析成像（Cryo-ET）的专家的合作继续为确定病毒和大分子结构的分析辅助提供至关重要的分析辅助，以及自动层析成像对齐，以提高数据库效率和工作流程。 新人员包括实习生和合同雇用，他们带来了显微镜和数据管理专业知识。 两种技术（即免疫标记，SEM SEM，蒙太奇层析成像）和技术（即直接电子检测器和伏尔泰相板）的持续发展都具有扩展的功能，并在3 Angstroms处有显着改善3D可视化和分辨率的潜力。 导致数据分析，数据管理，标本准备和数据收集的协作对于有效发展高度专业的技能非常宝贵。 流式细胞仪 在BSL-2和BSL-3环境中开发了特定于项目的研究应用程序，用于流式细胞仪分析和分类。 RML基因组学单元 RML基因组学单元为DIR研究人员提供了有关高通量基因组技术（例如下一代测序（NGS）和QPCR）的实验设计，样品制备，实验和数据分析的专业知识。 RML基因组学单元（GU）专门研究宿主和/或病原体的基因表达分析，细菌基因组的从头组装以及病原体基因组和人类外观的SNP/INDEL分析。 该单元还以高吞吐量进行了桑格测序，并协助研究人员对困难的测序区域进行故障排除。 这些单位的重点是帮助研究人员优化其对基因组技术的使用。通过了解实验设计和数据分析中的技术和经验，该单元可以确定样品和复制的理想数量，最佳的技术平台以及适当的分析，以最有效的方式实现所需的结果。 视觉和医学艺术 Rocky Mountain Laboratories（RML）的视觉和医学艺术部门（VMA）提供了高度专业化和先进的技术，以支持壁内研究（DIR）研究计划。该单元提供了各种各样的数字技术，涉及一套综合的插图，设计服务，摄影，3D动画，视频和新媒体，以记录科学发现。 此外，VMA向主任办公室，NIAID提供了科学沟通，向科学家，公众以及美国立法者提供了介绍。 视觉和医学艺术（VMA）单元与研究人员密切合作，提供最高质量的插图和图像，以帮助进行复杂的科学思想和发现的视觉交流。除了支持DIR的研究外，VMA还为美国国家过敏和传染病研究所（NIAID）办公室提供各种媒体支持。其中一些任务包括创建用于国会和外国贵宾证词中的视觉效果。