Inverted two photon laser scanning microscope for host defense

用于宿主防御的倒置双光子激光扫描显微镜

• 批准号: 7392075
• 负责人: Michael Loran Dustin
• 金额: $ 50万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7392075
• 关键词: Abscess; Address; Area; Autoimmunity; Cells; Contract Services; Depth; Devices; Equipment; Floor; Funding; Genus Mycobacterium; Gut associated lymphoid tissue; Host Defense; Housing; Human; Image; Immune; Immune response; Incidence; Infection; Inflammatory; Institutes; Intestines; Lasers; Life; Listeria; Liver; Microscope; Microscopic; Microscopy; Mus; Organ; Parasites; Pattern; Population; Principal Investigator; Public Health; Research Personnel; Scanning; Spleen; Structure of parenchyma of lung; System; T-Lymphocyte; Time; Tissues; Toxin; United States National Institutes of Health; Viral; Visceral; base; biosafety level 2 facility; experience; fighting; human tissue; instrument; instrumentation; intravital microscopy; lymph nodes; migration; novel; pathogen; reconstitution; two-photon

DESCRIPTION (provided by applicant): This is a proposal from a group of five NIH funded principal investigators requesting funds to purchase a commercially available inverted two-photon laser scanning microscope (TPLSM) system that will be primarily utilized for BSL2 projects. This shared instrument will provide new capabilities for deep tissue imaging in visceral organs (liver, spleen and intestines) in mice infected with BSL2 level pathogens. These new capabilities are essential for specific projects focusing on host-defense against bacterial, viral and parasitic attack. We propose to characterize the immune response to Listeria in live mouse spleen, lymph nodes and liver (Project 1), the immune response to Mycobacteria in the lymph nodes and lung tissue (Project 2), the immune response to human malarial parasites in reconstituted human tissue (Project 3), determine the migration patterns of inflammatory T cell populations in the gut associated lymphoid tissues during infection with bacterial and protozoan pathogens (Project 4), and determine the impact of Staphylococcal toxins on innate immune cells during abscess formation (Project 5). For each of the projects, capabilities unique to the new system will be invaluable. The key advantages of the new system compared to existing systems are 1) inverted configuration, which is ideal for intravital microscopy of the visceral organs, and 2) it will be housed in a BSL-2 facility. The instrument will be housed on the second floor of the Skirball institute where 3 of the major users and the facility manager are located. The facility manager has experience with operating BSL3 microscopy equipment and has over 30 years experience with laser based instrumentation. This device will have long-term institutional support to cover the service contracts. Time will also me made available to other NIH funded researchers at NYU and in the NY area who need the unique capabilities of the Leica SP5 TPLSM system. The inverted system is unusual for intravital microscopy systems, but extensive justification is provided for why the inverted system is needed for the imaging of tissues that are major targets for pathogen entry and colonization. The system will provide vital support for novel studies on host-defense and host- pathogen interactions. Hypotheses that require dynamic microscopic information about pathogen, host cell and tissue changes during infection can be addressed. The impact on public health will be to develop better strategies to fight infections and reduce the incidence of pathogen triggered autoimmunity.

描述（由申请人提供）：这是由 NIH 资助的五名主要研究人员组成的小组提出的提案，请求资金购买商用倒置双光子激光扫描显微镜 (TPLSM) 系统，该系统将主要用于 BSL2 项目。该共享仪器将为感染 BSL2 级病原体的小鼠内脏器官（肝脏、脾脏和肠道）的深层组织成像提供新功能。这些新功能对于专注于宿主防御细菌、病毒和寄生虫攻击的特定项目至关重要。我们建议表征活体小鼠脾脏、淋巴结和肝脏中对李斯特菌的免疫反应（项目 1）、淋巴结和肺组织中对分枝杆菌的免疫反应（项目 2）、重组人体中对人类疟疾寄生虫的免疫反应组织（项目 3），确定细菌和原生动物病原体感染期间肠道相关淋巴组织中炎症 T 细胞群的迁移模式（项目 4），并确定脓肿形成期间先天免疫细胞上的葡萄球菌毒素（项目 5）。对于每个项目来说，新系统独有的功能将是无价的。与现有系统相比，新系统的主要优点是 1) 倒置配置，非常适合内脏器官的活体显微镜检查，2) 它将安装在 BSL-2 设施中。该仪器将安装在 Skirball 研究所的二楼，其中 3 位主要用户和设施经理就位于该二楼。设施经理拥有操作 BSL3 显微镜设备的经验，并且拥有 30 多年使用激光仪器的经验。该设备将获得长期机构支持以涵盖服务合同。我还将为纽约大学和纽约地区其他需要 Leica SP5 TPLSM 系统独特功能的 NIH 资助研究人员提供时间。倒置系统对于活体显微镜系统来说是不寻常的，但是对于为什么需要倒置系统来对作为病原体进入和定植的主要目标的组织进行成像提供了广泛的理由。该系统将为宿主-防御和宿主-病原体相互作用的新研究提供重要支持。需要有关感染过程中病原体、宿主细胞和组织变化的动态微观信息的假设可以得到解决。对公共卫生的影响将是制定更好的策略来对抗感染并减少病原体引发的自身免疫的发生率。