Confocal upgrade for intravital microscopy following vascular injury

描述（由申请人提供）：血管损伤后活体显微共聚焦升级该提案的目标是使现有的共享仪器现代化，该仪器已经使用了5年，用于观察基因工程小鼠血管损伤的止血反应。止血是指在损伤部位迅速积聚循环血小板和纤维蛋白，从而限制失血。具有快速反应止血机制的意外后果是血小板/纤维蛋白凝块的不适当形成是心脏病发作和中风的主要原因，特别是在动脉粥样硬化的情况下。尽管在体外对这些事件已经有了很多了解，但直到最近才有可能在体内研究它们。止血和血栓形成研究的基础和临床研究是宾夕法尼亚大学医学院和费城儿童医院（CHOP）的主要重点。五年前，我们组装了一个（当时）最先进的仪器，使我们能够使用数字活体荧光显微镜观察微循环内的事件。从测试血小板活化的新想法到寻找治疗血友病的新方法，原始仪器已经在各种研究中得到了很好的应用。直接社区的成员以及来访的调查人员都分享了这份报告。最初组装时，在美国只有一种类似的仪器，即使现在也很少有其他的。然而，原来的仪器包括共聚焦功能，其用途被证明是有限的相对较长的时间（bbb45秒），需要捕捉一组完整的图像。更好，更快的共聚焦头现在是可用的，可以改装到我们现有的显微镜。申请的资金将用于升级共聚焦头、激光器和相机，并用速度快得多的波长变换器取代波长变换器。其最终效果将使我们能够更详细地观察事件，并具有更强的空间关系感，使我们回到回答生物学重要问题所需的最先进技术水平。我们知道这个新系统会像预期的那样工作，因为哈佛大学的Furie实验室最近完成了一个类似的仪器。虽然我们的具体生物学问题不同，但他们的经验对系统的能力提供了信息。该提案包括将受益于新仪器的美国国立卫生研究院资助的研究的例子，以及继续使用该仪器的管理计划。经济影响：宾夕法尼亚大学医学院和CHOP为当地经济做出了巨大贡献。2008年，它们创造并支持了54,000多个就业岗位和110亿美元的地区经济活动。目前的提案将帮助我们保持我们在竞争资助支持方面的优势，并在宾夕法尼亚大学创造或保留至少4个工作岗位，以及在智能成像创新方面的更多工作岗位，智能成像创新是我们开发和维护仪器的合作伙伴。1

DESCRIPTION (provided by applicant): Confocal upgrade for intravital microscopy following vascular injury 6. Project summary The goal of this proposal is to modernize an existing shared instrument that has been used for the past 5 years to observe the hemostatic response to vascular injury in genetically engineered mice. Hemostasis refers to the rapid accumulation of circulating platelets and fibrin at a site of injury, thereby limiting blood loss. The unintended consequence of having a rapid response hemostatic mechanism is that inappropriate formation of platelet/fibrin clots is a primary contributor to heart attacks and strokes, particular in the setting of atherosclerosis. Although much has been learned about these events in vitro, it has only recently been possible to study them in vivo. Basic and clinical research in hemostasis and thrombosis research is a major focus at Penn Medicine and Children's Hospital of Philadelphia (CHOP). Five years ago, we assembled a (then) state of the art instrument that has enabled us to use digital Intravital fluorescent microscopy to observe events within the microcirculation. The original instrument has been put to good use in studies that range from testing new ideas about platelet activation to efforts to finding novel ways to cure hemophilia. It has been shared by members of the immediate community as well as by visiting investigators. When originally assembled, there was only one comparable instrument in the United States and even now there are few others. However, the original instrument included confocal capabilities whose usefulness proved to be limited by the relatively long times (>45 sec) required to capture an entire set of images. Better, faster confocal heads are now available and can be retrofitted to our existing microscope. The requested funds will be used to upgrade the confocal head, lasers and camera, and replace the wavelength changer with one that is much faster. The net effect will be to allow us to observe events in greater detail and with a greater sense of spatial relationships, bringing us back to the level of state of the art technology needed to answer biologically-important questions. We know that the new system will work as expected, because a similar instrument was recently completed in the Furie lab at Harvard. Although our specific biological questions differ, their experience is informative about the capabilities of the system. This proposal includes examples of the NIH-funded research that will benefit from the new instrument, as well as a management plan for its continued use. Economic impact: Penn Medicine and CHOP contribute substantially to the local economy. In 2008, they created and supported more than 54,000 jobs and $11 billion in regional economic activity. The current proposal will help us to maintain our edge in competing for grant support and create or retain at least 4 jobs at Penn plus more at Intelligent Imaging Innovations, our partner in developing and maintaining the instrument. 1