In Vito Imaging

多光子显微镜是一种技术，涉及成像的荧光标记的细胞内完整的 组织使用激光扫描显微镜和脉冲红外激光。这种方法可以用于 无论是切除大块组织或活的，麻醉的动物，一种技术称为活体显微镜 （IVM）。在拟议的YRDRCC中，许多研究人员认为，将这些技术应用于他们的研究， 研究将允许快速进展，并为他们的体内生物学问题提供实质性的见解 这在传统的体外分析中是不可能的。然而，为了执行该技术， 除了少数调查人员之外，需要昂贵得令人望而却步的设备，即激光扫描仪。 显微镜（LSM）和多光子（MP）激光。耶鲁大学医学院的研究人员正在 幸运的是，学校已经投资了多光子激光器，并购买了额外的 定制设计的激光扫描显微镜，配备灵敏的外部探测器， 收集低水平光。一个独特的显微镜套件正在建设中，将允许 动物的准备和随后的成像在相邻的房间进行。这个显微镜系统 将提供给耶鲁大学的研究人员，包括YRDRCC的成员。但许多 目前存在的障碍阻止研究者将这种方法用于他们的体内研究。为 例如，MP激光器的用户成本明显高于显微镜本身的成本， 这主要是由于维护MP激光器的服务合同。所需的额外技能 大多数研究人员似乎无法在IVM期间获得动物麻醉、手术和约束， 在免疫学中，相对较少的研究者在他们的研究中使用了IVM， 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，而这些蛋白很难及时获得 时尚.因此，本申请建议建立一个体内Imagng核心设施， 减少这些障碍，而且还提供深层组织和活体动物成像方面的培训和建议。 拟议中的IVICF将对细胞间相互作用的研究步伐产生巨大影响， 在健康和风湿性和自身免疫性疾病中的体内分化和迁移。

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.