FLUORESCENCE LIFETIME IMAGING MICROSCOPY SYSTEM

描述(由申请人提供)：我们正在申请资金，以使用PicoQuant的系统特定升级套件将我们位于光学成像核心(OIC)的奥林巴斯FV1000共焦显微镜升级到完全功能的荧光寿命(FLT)成像显微镜(FLIM)。Flim测量细胞微环境中的Flt。与荧光强度不同的是，通过对激发态性质的微扰，Flt对局部探测环境非常敏感。位于圣路易斯的华盛顿大学(WUSTL)和周围的研究界无法使用FLIM系统，该系统提供易于使用的界面，并执行具有在多个通道中检测荧光的能力的不同波长成像。FLIM系统将支持17个以上由NIH资助的项目，最初包括来自WUSTL、圣路易斯大学(SLU)和密苏里大学圣路易斯分校(UMSL)的3个主要用户和11个次要用户。所有的研究人员都发表了大量的分子成像方面的工作，并证明了对薄膜的持续需求。计划中的研究将集中在发现导致报告分子Flt变化的新的分子相互作用，设计新型Flt造影剂，定量确定目标生物分子的亚细胞分布，建立不同荧光分子探针和抗癌药物在细胞间的行为，了解细胞中大分子的复杂动力学，解开未知功能蛋白质的生化作用，并阐明在高分辨率胶片的非侵入性小动物成像研究中观察到的Flt异质性的生物学意义。该系统不仅可以进行Flt和常规的荧光强度测量，还可以提供目标生物分子或过程的时空信息，以响应细胞微环境的变化，如pH、亲脂性、细胞代谢状态、自由基的存在和缺氧。拟议的系统升级将加强OIC提供的生物过程分子成像服务，并推动圣路易斯许多不断发展的研究小组的研究。它将促进新的合作并产生新的概念，以促进我们在细胞水平上对复杂生物系统的理解。由于OIC位于医疗中心的中心位置，该系统将向所有当前和未来的用户开放，就像目前OIC中安装的其他仪器一样。经验丰富的人员将管理仪器，培训新用户，并帮助实施新项目。该仪器靠近威斯康星科技大学、南加州大学和密歇根州立大学的研究实验室，将吸引新的用户，并利用最先进的技术平台促进对学生和研究员的培训。

DESCRIPTION (provided by applicant): We are requesting funding to upgrade our Olympus FV1000 confocal microscope at the Optical Imaging Core (OIC) to a fully capable fluorescence lifetime (FLT) imaging microscope (FLIM) using system-specific upgrade kit from PicoQuant. FLIM measures FLT in cell microenvironment. Unlike fluorescence intensity, FLT is exquisitely sensitive to local probe environment through perturbation of excited state properties. Washington University in St. Louis (WUSTL) and surrounding research community do not have access to a FLIM system that provides easy-to-use interface and performs diverse wavelength imaging with the capability of detecting fluorescence in multiple channels. The FLIM system will support 17+ NIH-funded projects, initially consisting of 3 major and 11 minor users from WUSTL, Saint Louis University (SLU), and University of Missouri at St. Louis (UMSL). All the investigators have substantially published work in molecular imaging and demonstrable ongoing need for FLIM. Planned studies will focus on discovering novel molecular interactions that result in changes in the FLT of reporter molecules, designing new class of FLT contrast agents, quantitatively determining the sub-cellular distribution of target biomolecules, establishing the behavior of different fluorescent molecular probes and cancer drugs in cell compartments, understanding the complex dynamics of macromolecules in cells, unraveling the biochemical roles of proteins of unknown functions, and elucidating the bio-significance of FLT heterogeneity observed in noninvasive small animal imaging studies with high resolution FLIM. Not only will this system perform FLT and conventional fluorescence intensity measurement, it will also provide spatio-temporal information of a target biomolecule or process in response to cell microenvironmental changes stimulated by factors such as pH, lipophilicity, cell metabolic state, presence of free radicals, and hypoxia. The proposed system upgrade will enhance the services available in OIC for molecular imaging of biological processes and advance the research of many growing research groups at St. Louis. It will facilitate new collaborations and generate new concepts to advance our understanding of complex biological systems at cell level. Because of OIC's location in the heart of the medical center, the system will be available to all current and future users, as is the current practice with other instruments housed in OIC. Highly experienced personnel will manage the instrument, train new users, and help conduct new projects. The proximity of the instrument to research laboratories at WUSTL, SLU, and UMSL will attract new user and facilitate training of students and fellows using the state-of-the-art technology platform.