Olympus BX-DSU Live-Cell Disk Scanning Imaging / Electrophysiology System

奥林巴斯 BX-DSU 活细胞盘扫描成像/电生理学系统

• 批准号: 7794331
• 负责人: BRADLEY K. TAYLOR
• 金额: $ 29.71万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-13 至 2011-05-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7794331
• 关键词: Address; Auditory; Brain Injuries; Calcium; Cardiac; Cardiovascular Diseases; Cells; Confocal Microscopy; Detection; Disease; Dyes; Electrophysiology (science); Environment; Fura-2; Generations; Grant; Heart; Image; Ion Channel; Kentucky; Lasers; Life; Light; Lung diseases; Maintenance; Microscope; Nervous system structure; Neurosciences; Obesity; Paper; Pharmaceutical Preparations; Photobleaching; Physiology; Publishing; Resolution; Scanning; Sensory; Signal Transduction; Slice; Solutions; Source; System; Techniques; Technology; Tissues; Universities; Wages; Work; cellular imaging; charge coupled device camera; chronic pain; cost; experience; flash photolysis; flexibility; instrument; instrumentation; olfactory disorder; patch clamp; photolysis; ratiometric; relating to nervous system; response

DESCRIPTION (provided by applicant): The long-term objectives of our 12 federal grants (R01-type) are to address important and unique problems in neuroscience and cardiac physiology, including a better understanding how living cells in the nervous system and heart respond to drugs or other changes in the environment. The current application is in response to a significant need of our 8 Major Users to obtain confocal images and electrophysiological recordings, often simultaneously, from live neural, sensory and cardiac cells or tissue slices. Specific needs include the following major functional capabilities: Confocal imaging of Ca2+ transients and fluorescent markers in live tissue slices; ratiometric Fura-2 imaging of Ca2+ transients; high temporal and spatial resolution of Ca2+ imaging during concomitant patch clamp recordings of ion channel activity; and UV Flash photolysis of caged compounds. All of the Major Users have extensive experience with live cell calcium imaging, electrophysiology, and/or confocal microscopy. Collectively, we have published over a hundred papers using one or more of these techniques, including 15 papers that simultaneously used functional confocal imaging and electrophysiological recording in live cells. An inherent problem of laser confocal microscopes is the very intense excitation light, leading to substantial dye decomposition (photobleaching) during live-cell imaging. The proposed Olympus BX- DSU Live-Cell Disk Scanning Imaging System overcomes this problem with optimization of signal detection. It uses spinning disk confocal technology with a new generation, super-cooled EM-CCD camera to generate images with high temporal and spatial resolution. The broad spectrum light source produces substantially less photobleaching than do lasers. Furthermore, this system adds a UV flash illuminator for photolysis of photoactivatable (caged) compounds, complete integration of patch-clamp instrumentation for concomitant electrophysiological recordings, and an upright stand preferred for tissue slice work. The result is a very cost-effective and flexible solution that responds to all described needs of the Major Users. No other instrument at or near the University of Kentucky can provide this constellation of features. The University will provide appropriate space, salary support for experienced technical staff, and maintenance for the system. The system will be set up within a large well-established Imaging Facility at the University of Kentucky by experts in scanning disk technology and electrophysiology. This system will allow the Major Users to continue to achieve new discoveries towards the better understanding and treatment of chronic pain, obesity, auditory disorders, olfactory disorders, pulmonary disease, consequences of brain injury, and cardiovascular disease.

描述（由申请人提供）：我们的12个联邦补助金（R01型）的长期目标是解决神经科学和心脏生理学中的重要和独特的问题，包括更好地了解神经系统中的活细胞如何对毒品或环境中的其他变化。当前的应用是对我们的8个主要用户的重大需求，以获取共聚焦图像和电生理记录，通常是同时发生的，来自活神经，感觉和心脏细胞或组织切片。特定需求包括以下主要功能功能：在活组织切片中的Ca2+瞬态和荧光标记的共聚焦成像； Ca2+瞬变的比例fura-2成像；在离子通道活性的贴片夹记录过程中，Ca2+成像的高时间和空间分辨率；和蛋白质化合物的紫外线闪光光解。所有主要用户在活细胞成像，电生理学和/或共聚焦显微镜方面都有丰富的经验。总的来说，我们使用其中一种或多种技术发表了一百多篇论文，其中包括同时在活细胞中使用功能共聚焦成像和电生理记录的15篇论文。激光共聚焦显微镜的固有问题是非常强烈的激发光，导致活细胞成像过程中实质性的染料分解（光漂白）。提出的Olympus BX-DSU Live细胞磁盘扫描成像系统通过优化信号检测来克服这个问题。它使用新一代，超冷的EM-CCD摄像头旋转磁盘共聚焦技术来生成具有高时间和空间分辨率的图像。与激光器相比，宽光谱光源产生的光漂白要少得多。此外，该系统添加了一个用于光分解光含量（笼中）化合物的紫外线照明器，将贴片钳仪器完整整合用于伴随电生理记录，以及用于组织切片工作的直立支架。结果是一个非常具有成本效益且灵活的解决方案，可满足主要用户的所有描述需求。肯塔基大学或附近没有其他乐器可以提供这一特征。大学将为经验丰富的技术人员提供适当的空间，薪水支持以及对系统的维护。该系统将通过扫描磁盘技术和电生理学专家在肯塔基大学的大型成像设施中建立。该系统将使主要用户能够继续获得新的发现，以更好地了解和治疗慢性疼痛，肥胖，听觉疾病，嗅觉疾病，肺部疾病，脑损伤后果和心血管疾病。