Total Internal Reflection Fluorescence Microscopy System

描述(申请人提供)：此申请是为了支持购买徕卡AM TIRF(全内反射荧光)显微镜系统。该仪器将被安置在托马斯·杰斐逊大学坎摩尔癌症中心的设施内。该设施为基梅尔癌症中心、整个托马斯·杰斐逊大学的用户提供光学显微镜服务，并根据需要向邻近研究机构的研究人员提供服务。托马斯杰斐逊大学目前没有TIRF设施。在TIRF模式下，所要求的仪器将能够成像质膜结构、进出质膜的囊泡运输和其他膜活动，以及具有异常高灵敏度和低本底的单分子。TIRF照明由灵敏的广域荧光成像补充，这也将提供出色的FRET能力。这些属性配备了多条激光线路和快速的AOTF波长切换，非常适合用户对活细胞快速成像的需求。TIRF系统对于支持来自托马斯杰斐逊大学五个系(生物化学和分子生物学、癌症生物学、医学、微生物学和免疫学以及生理学)和两个研究中心(基梅尔癌症中心、转化医学中心)的八个主要用户的研究项目至关重要。所有选定的主要用户都有正在进行的、资金充足的研究计划，以及在活细胞和固定细胞的荧光和共聚焦显微镜方面的经验和专业知识。此外，该仪器还将服务于基梅尔癌症中心内对TIRF显微镜感兴趣的其他临时用户，以及托马斯·杰斐逊大学的其他研究人员。与公共健康相关：质膜是细胞与其环境之间信息和材料传递的关键界面。本建议书中要求的徕卡AM TIRF显微镜是唯一专门用于研究这一至关重要的细胞领域的仪器，使用该仪器进行的研究将为包括癌症、心血管疾病、神经疾病和传染病在内的各种疾病提供重要的洞察力。这些疾病领域包括托马斯·杰斐逊大学的主要研究优势和长期生物医学目标。

DESCRIPTION (provided by applicant): This application is for support to purchase a Leica AM TIRF (total internal reflection fluorescence) microscope system. The instrument will be housed within the - Facility of the Kimmel Cancer Center of Thomas Jefferson University. This Facility provides light microscopy services to users within the Kimmel Cancer Center, throughout Thomas Jefferson University, and as needed and available to investigators at neighboring research institutions. There is no TIRF facility presently available at Thomas Jefferson University. In TIRF mode, the requested instrument will provide the ability to image plasma membrane structures, vesicle transport to and from the plasma membrane and other membrane activities, as well as single molecules with exceptionally high sensitivity and low background. The TIRF illumination is complemented by sensitive widefield epifluorescence imaging, which will also provide excellent FRET capability. Equipped with multiple laser lines and fast AOTF wavelength switching, these attributes are uniquely suited to the users' needs for rapid imaging of live cells. The TIRF system is vitally needed to support the research programs of eight major users drawn from five Departments (Biochemistry and Molecular Biology, Cancer Biology, Medicine, Microbiology and Immunology, and Physiology) and two Research Centers (Kimmel Cancer Center, Center for Translational Medicine) of Thomas Jefferson University. All of the selected major users have ongoing and well-funded research programs, as well as experience and expertise with fluorescence and confocal microscopy of live and fixed cells. In addition, the instrument will serve other more occasional users within the Kimmel Cancer Center who have expressed interest in TIRF microscopy, as well as other research investigators of Thomas Jefferson University. PUBLIC HEALTH RELEVANCE: The plasma membrane is the interface that is key to the transfer of information and material between the cell and its environment. The Leica AM TIRF microscope requested in this proposal is uniquely specialized for investigating this critically important cellular domain, and studies carried out with this instrument will provide important insight into a variety of diseases including cancer, cardiovascular disease, neurological disorders, and infectious diseases. These disease areas encompass the major research strengths and long-range biomedical goals of Thomas Jefferson University.