In Vivo Imaging of Borrelia Burgdorferi Infection

多光子显微镜是一种对完整细胞内荧光标记的细胞进行成像的技术。 使用激光扫描显微镜和脉冲红外线激光扫描组织。此方法可用于 要么是切除的大块组织，要么是活的麻醉动物，这项技术被称为活体显微镜 (IVM)。拟议中的青藏铁路研究中心内的许多调查人员认为，将这些技术应用于他们的 研究将允许快速进展，并为他们在体内的生物学问题提供实质性的见解 以一种传统的体外分析不可能实现的方式。然而，为了执行该技术， 除了少数调查人员外，所需的设备都贵得令人望而却步，即激光扫描 显微镜(LSM)和多光子(MP)激光。耶鲁大学医学院的调查人员 幸运的是，学校已经投资了多光子激光器，并额外购买了一台 定制设计的激光扫描微槽配备了灵敏的外部探测器，允许 低亮度光的集合。一个独特的显微镜套件现在正在建设中，它将允许 准备动物及其随后在相邻房间进行的成像。这套显微镜系统 将提供给耶鲁大学的调查人员，包括YRDRCC的那些成员。然而，许多人 目前存在的障碍阻碍了研究人员利用这种方法进行体内研究。为 例如，MP激光器的用户成本显著高于显微镜本身的成本， 这在很大程度上是因为维护MP激光器的服务合同。所需的其他技能 对于大多数研究人员来说，在IVM期间进行动物麻醉、手术和约束似乎是无法获得的，因为 免疫学领域的研究人员相对较少，在他们的研究中使用IVM，因此可以使用 为更广泛的免疫学社区提供培训和建议。同样，体内荧光的检测 通常需要表达荧光蛋白的小鼠品系，而这些荧光蛋白很难及时获得 时尚。因此，该应用程序建议建立一个活体成像核心设施，不仅 减少这些障碍，但也提供深层组织和活体动物成像方面的培训和建议。 拟议的试管内受精将对细胞间相互作用的研究速度产生重大影响， 在健康、风湿病和自身免疫性疾病中体内的分化和迁移。

Multiphoton microscopy is a technique that involves the imaging of fluorescently tagged cells within intact tissue using laser scanning microscopes and pulsed infra-red lasers. This approach can be employed with either excised bulk tissue or with live, anesthetized animals, a technique referred to as intravital microscopy (IVM). Many investigators within the proposed YRDRCC feel that the application of these techniques to their research would allow for rapid progress and provide substantial insights into their in vivo biological questions in a way that is not possible with traditional in vitro assays. However, in order to perform this technique, equipment is required that is prohibitively expensive to all but a few investigators, namely a laser scanning microscope (LSM) and multiphoton (MP) laser. Yale University School of Medicine investigators are fortunate in that the school has already invested in the multiphoton laser and has purchased an additional custom designed laser scanning microsocpe equipped with sensitive external detectors that allow for the collection of low level light. A unique microscopy suite is now under construction that will permit the preparation of animals and their subsequent imaging to occur in adjacent rooms. This microscopy system will be available to Yale investigators, including those members of the YRDRCC. Nevertheless, many barriers currently exist that prevent investigators from utilizing this approach to their in vivo research. For example, the cost to users of the MP laser is significantly higher than the cost of the microscope by itself, largely due to the service contracts for maintenance of the MP laser. The additional skills needed for the animal anesthesia, surgery and restraint during IVM appear unobtainable to most researchers, given that relatively few investigators within immunology have employed IVM in their research and are thus available for training and advice to the broader immunology community. Likewise, the detection of fluorescence in vivo often requires strains of mice expressing fluorescent proteins that can be difficult to obtain in a timely fashion. Therefore, this application proposes to establish an In Vivo Imagng Core Facility that would not only reduce these barriers but also provide training and advice in the imaging of deep tissue and live animals. The proposed IVICF would have a dramatic impact on the pace of research into cell-cell interactions, differentiation and migration in vivo in both health and in rheumatic and autoimmune diseases.