Inverted two photon laser scanning microscope for host defense

描述（由申请人提供）：这是一个由五名NIH资助的主要研究者组成的小组提出的建议，要求提供资金，以购买一个商业上可用的倒置双光子激光扫描显微镜（TPLSM）系统，该系统将主要用于BSL 2项目。该共享仪器将为感染BSL 2级病原体的小鼠的内脏器官（肝、脾和肠）的深层组织成像提供新的能力。这些新功能对于专注于针对细菌、病毒和寄生虫攻击的主机防御的特定项目至关重要。我们建议在活小鼠脾脏、淋巴结和肝脏中表征对李斯特菌的免疫应答（项目1），淋巴结和肺组织中对分枝杆菌的免疫反应（项目2），重组人体组织中对人类疟疾寄生虫的免疫反应（项目3），确定细菌和原生动物感染期间肠道相关淋巴组织中炎性T细胞群的迁移模式病原体（项目4），并确定脓肿形成过程中葡萄球菌毒素对先天免疫细胞的影响（项目5）。对于每个项目，新系统特有的能力将是无价的。与现有系统相比，新系统的主要优势是1）倒置配置，这是内脏器官活体显微镜的理想选择，2）它将被安置在BSL-2设施中。该仪器将安置在Skirball研究所的二楼，3个主要用户和设施管理员都在那里。该设施经理拥有操作BSL 3显微镜设备的经验，并拥有超过30年的激光仪器经验。该设备将得到长期的机构支持，以涵盖服务合同。时间也将提供给其他NIH资助的研究人员在纽约大学和纽约地区谁需要的徕卡SP 5 TPLSM系统的独特功能。倒置系统是不寻常的活体显微镜系统，但提供了广泛的理由，为什么需要倒置系统的病原体进入和定植的主要目标的组织成像。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。假设，需要动态微观信息的病原体，宿主细胞和组织的变化，在感染过程中可以解决。对公共卫生的影响将是制定更好的战略来对抗感染，减少病原体引发的自身免疫的发生率。

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.