In Vito Imaging

活体成像

• 批准号: 7352530
• 负责人: Mark J Mamula
• 金额: $ 13.18万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-17 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7352530
• 关键词: Anesthesia procedures; Animals; Autoimmune Diseases; Backcrossings; Biological; Cell Communication; Cells; Collection; Communities; Consult; Contract Services; Core Facility; Cost Analysis; Custom; Data; Depth; Detection; Educational workshop; Elements; Equipment; Experimental Designs; Facility Construction Funding Category; Fluorescence; Fluorochrome; Funding; Future; Health; Image; Image Analysis; Imaging Techniques; Immunologist; Immunology; Individual; Knowledge; Label; Lasers; Life; Light; Maintenance; Microscope; Microscopy; Mouse Strains; Operative Surgical Procedures; Photons; Physiologic pulse; Preparation; Proteins; Pulse taking; Research; Research Personnel; Resolution; Rheumatism; Rheumatology; Scanning; Schools; Speed; Structure; Surveys; System; Techniques; Time; Tissues; Training; Translations; Universities; biological research; cost; density; design; detector; in vitro Assay; in vivo; insight; interest; intravital microscopy; medical schools; member; migration; prevent; programs; reconstruction; research study; restraint; skills

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.

多光子显微镜是一种涉及完整内荧光标记的细胞成像的技术 使用激光扫描显微镜和脉冲红外激光器组织。可以使用这种方法 切除的散装组织或活着的麻醉动物，这是一种被称为插入显微镜的技术 （IVM）。拟议的YRDRCC中的许多调查人员认为，这些技术在其 研究将允许快速进步，并为其体内生物学问题提供大量见解 在传统的体外测定中是不可能的。但是，为了执行此技术， 需要设备对除一些调查人员以外的所有人都非常昂贵，即激光扫描 显微镜（LSM）和多光子（MP）激光。耶鲁大学医学院调查人员是 幸运的是，学校已经投资于多光子激光器，并购买了另外 定制设计的激光扫描微孔，配备了敏感的外部探测器，可允许 低水平光的收集。现在正在建造一个独特的显微镜套件，将允许 动物的制备及其随后发生的成像发生在相邻的房间中。这个显微镜系统 耶鲁大学的调查人员将使用，包括YRDRCC的那些成员。尽管如此，很多 目前存在的障碍可以阻止研究人员利用这种方法来进行体内研究。为了 例如，MP激光器用户的成本明显高于显微镜的成本， 很大程度上是由于维护MP激光的服务合同。所需的其他技能 鉴于这一点 免疫学中相对较少的研究人员在研究中使用了IVM，因此可以使用 向更广泛的免疫学社区进行培训和建议。同样，在体内检测荧光 通常需要表达荧光蛋白的小鼠菌株，这些蛋白很难及时获得 时尚。因此，本应用程序建议建立一个体内Imagng核心设施，不仅将 减少这些障碍，但还提供了深层组织和活体动物的成像的培训和建议。 拟议的IVICF将对细胞 - 细胞相互作用的研究速度产生巨大影响， 健康，风湿性和自身免疫性疾病的体内分化和迁移。