INTRAVITAL VIDEO-RATE CONFOCAL MICROSCOPE

活体视频共焦显微镜

• 批准号: 6288065
• 负责人: Bruce Furie
• 金额: $ 17.54万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-15 至 2002-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6288065
• 关键词: biomedical equipment purchase; confocal scanning microscopy

This application is submitted for a video-rate confocal microscopy facility for intravital microscopy in living small. animals, such as genetically altered mice, in living cells rolling in a flow chamber and viable tissue maintained in tissue culture. We wish to merge three advanced technologies to develop our understanding of vascular disease, skin disease, thrombosi, cell signalling and cell: (1) intravital microscopy of blood vessels in living mice to study the impact of various perturbations of the vascular system on endothelium, leukocytes, platelets and plasma proteins including the blood clotting proteins; (2) the generation of mice altered by the knockout strategy or rendered transgenic, with the net effect of creating a protein deficiency or introducing a fluorophore into a protein of interest; (3) the development of video speed confocal microscopes, such as the BioRad RTS2000 and Noran OZ confocal scanning laser microscopes, that permit rapid, video rate, real-time data acquisition at high resolution. A consortium of investigators at Harvard Medical School has been established that have a special need for a video rate confocal microscope facility. This group is composed of a group of junior and established investigators who have a special need for video-rate confocal microscopy as it can be applied to their NIH-funded research programs. No such instrumentation is currently available at Harvard Medical School or its affiliated hospitals. The video-rate confocal microscopy system must include the following critical features: (1) An intravital microscopy facility for the use of live animals; (2) A laser confocal microscope system capable of rapid scanning and suitable for DIC and fluorescence microscopy under flow conditions; (3) A computer workstation with imaging software for image acquisition and reconstruction; (4) Multiple fluorescent channels for co- localization experiments. The BioRad RTS2000 real-time laser confocal system, fitted to a Nikon E600FN microscope, will provide a group of investigators with instrumentation to significantly advance their fields.

该申请是针对用于小型活体活体显微镜检查的视频速率共焦显微镜设备而提交的。动物，例如基因改造的小鼠，在流动室中滚动的活细胞和组织培养物中维持的活组织中。我们希望融合三项先进技术，以加深我们对血管疾病、皮肤病、血栓形成、细胞信号传导和细胞的理解：（1）活体小鼠血管活体显微镜检查，研究血管系统的各种扰动对内皮、白细胞、血小板和血浆蛋白（包括凝血蛋白）的影响； (2) 通过敲除策略改变或转基因的小鼠的产生，其净效应是产生蛋白质缺陷或将荧光团引入感兴趣的蛋白质中； (3) 视频速度共焦显微镜的开发，例如 BioRad RTS2000 和 Noran OZ 共焦扫描激光显微镜，可实现快速、视频速率、高分辨率的实时数据采集。哈佛医学院成立了一个研究人员联盟，他们对视频速率共焦显微镜设施有特殊需求。该小组由一群初级和资深研究人员组成，他们对视频速率共聚焦显微镜有特殊需求，因为它可以应用于 NIH 资助的研究项目。哈佛医学院或其附属医院目前没有此类仪器。视频速率共焦显微镜系统必须包括以下关键特征： (1) 用于活体动物的活体显微镜设施； （2）能够快速扫描并适用于流动条件下的DIC和荧光显微镜的激光共焦显微镜系统； (3) 具有用于图像采集和重建的成像软件的计算机工作站； (4)用于共定位实验的多个荧光通道。 BioRad RTS2000 实时激光共焦系统安装在 Nikon E600FN 显微镜上，将为一组研究人员提供仪器，以显着推进他们的领域。