Confocal upgrade for intravital microscopy following vascular injury

血管损伤后活体显微镜共聚焦升级

• 批准号: 7792722
• 负责人: LAWRENCE F BRASS
• 金额: $ 26.93万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2012-06-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7792722
• 关键词: Arts; Atherosclerosis; Back; Biological; Blood Platelets; Blood Vessels; Clinical Research; Coagulation Process; Communities; Economics; Event; Fibrin; Funding; Genetically Engineered Mouse; Goals; Grant; Head; Hemophilia A; Hemorrhage; Hemostatic Agents; Hemostatic function; Image; In Vitro; Injury; Lasers; Learning; Medicine; Microcirculation; Microscope; Microscopy; Myocardial Infarction; Occupations; Pediatric Hospitals; Philadelphia; Platelet Activation; Research; Research Personnel; Site; Stroke; System; Technology; Testing; Thrombosis; Time; United States; Visit; Work; digital; economic impact; experience; in vivo; innovation; instrument; intravital microscopy; member; novel; response; spatial relationship

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

描述(由申请人提供)：血管损伤后活体显微镜的共聚焦升级6.项目摘要这项建议的目标是使现有的共用仪器现代化，该仪器在过去5年中一直用于观察基因工程小鼠对血管损伤的止血反应。止血指的是循环中的血小板和纤维蛋白在损伤部位迅速积累，从而限制失血。具有快速反应止血机制的意外后果是，不适当的血小板/纤维蛋白凝块的形成是心脏病发作和中风的主要因素，特别是在动脉粥样硬化的背景下。虽然在体外对这些事件已经了解了很多，但直到最近才有可能在体内对它们进行研究。止血和血栓形成研究的基础和临床研究是宾夕法尼亚医学和费城儿童医院(CHOP)的主要重点。五年前，我们组装了一台(当时)最先进的仪器，使我们能够使用数字活体荧光显微镜来观察微循环内的事件。最初的仪器已经在一系列研究中得到了很好的利用，从测试关于血小板激活的新想法到努力寻找治疗血友病的新方法。直属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，美国只有一种可比的仪器，即使现在也几乎没有其他仪器了。然而，最初的仪器包括共焦功能，其有效性被证明受到捕获整个图像所需的相对较长的时间(&gt；45秒)的限制。现在有了更好、更快的共焦头，可以改装到我们现有的显微镜上。申请的资金将用于升级共焦头、激光和相机，并用速度快得多的波长转换器取代。其最终效果将是让我们能够更详细地观察事件，并更好地了解空间关系，将我们带回回答具有生物重要性的问题所需的最先进技术水平。我们知道，新系统将会像预期的那样工作，因为最近在哈佛的Furie实验室完成了一台类似的仪器。尽管我们的具体生物学问题不同，但他们的经验是关于系统能力的信息。该提案包括将受益于新仪器的由NIH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：宾夕法尼亚医学和CHOP为当地经济做出了巨大贡献。2008年，它们创造和支持了54,000多个就业机会和110亿美元的区域经济活动。目前的提案将有助于我们在竞争拨款支持方面保持优势，并在宾夕法尼亚大学创造或保留至少4个工作岗位，以及我们在开发和维护仪器方面的合作伙伴智能成像创新公司的更多工作岗位。1