Super-resolution confocal microscope

超分辨率共焦显微镜

• 批准号: 9274885
• 负责人: Klaus F. Ley
• 金额: $ 56.63万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-19 至 2018-04-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9274885
• 关键词: 10 year old; Atherosclerosis; Biological Assay; Cells; Collection; Data; Detection; Devices; Disease; Fluorescence; Fluorescence Recovery After Photobleaching; Fluorescence Resonance Energy Transfer; Funding; Hypersensitivity; Image; Immune; Immunofluorescence Immunologic; Immunology; Infection; Institutes; Insulin-Dependent Diabetes Mellitus; Journals; Lasers; Microscope; NIH Grants and Contracts; Nature; Noise; Principal Investigator; Proteins; Protocols documentation; Publishing; Research Institute; Research Personnel; Research Project Grants; Resolution; Rheumatoid Arthritis; Scanning; Science; Scientist; Signal Transduction; System; Technology; Tissues; United States National Institutes of Health; Visible Radiation; Work; Zika Virus; base; behavioral study; detector; instrument; intravital imaging; novel; photoactivation; receptor

ABSTRACT The La Jolla Institute for Allergy and Immunology (LJI) is a premier immunology research institute in the US whose 24 principal investigators are supported by ~$40 million in NIH grants and contracts. At LJI we study the behavior of immune cells in diseases prominent in the US and around the world, including: type 1 diabetes, atherosclerosis, rheumatoid arthritis, and Zika infection. Nearly all of our scientists use laser scanning confocal imaging to visualize cellular interactions, signaling, and function and the results have been published in Nature, Science, and other prominent journals. However, to progress further in this important work, we need a versatile confocal microscope with super-resolution capabilities. Currently, LJI lacks a dynamic, capable, and technologically current confocal system and needs one that will yield the most impactful data for each NIH research project. This need includes a host of fluorescence based assays such as quantitative receptor translocation, protein co-localization intracellularly and in tissue, fluorescence resonance energy transfer (FRET), and fluorescence recovery after photobleaching (FRAP), and photoactivation. We have a 10 year old Leica SP5 combined confocal and multiphoton system that is dedicated to intravital imaging and a simple automated Olympus FV10i confocal suitable for fixed cells and tissues at lower resolutions. LJI has no super-resolution capability. Here, we seek support for purchasing a state-of-the-art Zeiss 880 confocal microscope with an Airyscan super-resolution detector, highly sensitive detection devices (GaAsP detectors), the greatest signal-to-noise ratio system for both fixed and live acquisition, multiple laser lines covering the entire visible light spectrum, and a fully tunable spectral emission collection grating. Airyscan is a novel super-resolution technology that can achieve 140 nm resolution using standard immunofluorescence protocols. After comparing all instruments on the market, it is clear that the Zeiss LSM 880 system is best suited for the needs of LJI investigators and, if funded, it will undoubtedly have a significant impact on NIH-supported scientific progress.

摘要 拉霍亚过敏和免疫学研究所（LJI）是一个首屈一指的免疫学研究机构， 美国的一个研究所，其24名主要研究人员得到了大约4000万美元的NIH赠款的支持， 合同.在LJI，我们研究免疫细胞在美国和周边地区突出疾病中的行为。 1型糖尿病、动脉粥样硬化、类风湿性关节炎和寨卡病毒感染。我们几乎所有的 科学家们使用激光扫描共聚焦成像来可视化细胞的相互作用、信号传导和功能， 研究结果已发表在《自然》、《科学》和其他著名期刊上。然而，为了在 这项重要的工作，我们需要一个多功能的共焦显微镜与超分辨率的能力。 目前，LJI缺乏一个动态的，有能力的，技术上最新的共焦系统，需要一个 将为每个NIH研究项目产生最有影响力的数据。这种需要包括大量的荧光 基于测定如定量受体易位，细胞内和组织中的蛋白共定位， 荧光共振能量转移（FRET）和光漂白后荧光恢复（FRAP），以及 光活化我们有一个10岁的徕卡SP5组合共聚焦和多光子系统， 专门用于活体成像和简单的自动化Olympus FV10i共聚焦，适用于固定细胞， 低分辨率的组织。LJI没有超分辨率能力。在这里，我们寻求支持购买 最先进的Zeiss 880共聚焦显微镜，配备Airyscan超分辨率探测器， 灵敏的探测器件（GaAsP探测器），最大的信噪比系统， 和实时采集，多条激光线覆盖整个可见光谱， 光谱发射收集光栅。Airyscan是一种新颖的超分辨率技术，可以实现140 nm分辨率。在比较了市场上的所有工具后， 很明显，蔡司LSM 880系统最适合LJI调查人员的需求，如果获得资助，它将 无疑对NIH支持的科学进步产生了重大影响。

项目成果

Structure of the rabies virus glycoprotein trimer bound to a prefusion-specific neutralizing antibody.

• DOI: 10.1126/sciadv.abp9151
• 发表时间: 2022-06-17
• 期刊: Science advances
• 影响因子: 13.6

Spatial and functional arrangement of Ebola virus polymerase inside phase-separated viral factories.

• DOI: 10.1038/s41467-023-39821-7
• 发表时间: 2023-07-13
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Fang, Jingru;Castillon, Guillaume;Phan, Sebastien;McArdle, Sara;Hariharan, Chitra;Adams, Aiyana;Ellisman, Mark H.;Deniz, Ashok A.;Saphire, Erica Ollmann
• 通讯作者: Saphire, Erica Ollmann

Proximity interactome analysis of Lassa polymerase reveals eRF3a/GSPT1 as a druggable target for host-directed antivirals.

• DOI: 10.1073/pnas.2201208119
• 发表时间: 2022-07-26
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1