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

描述（由申请人提供）：我们的12个联邦赠款（R 01型）的长期目标是解决神经科学和心脏生理学中重要和独特的问题，包括更好地了解神经系统和心脏中的活细胞如何对药物或环境中的其他变化做出反应。目前的应用是为了满足我们8个主要用户的重要需求，即从活的神经、感觉和心脏细胞或组织切片中同时获得共聚焦图像和电生理记录。具体需求包括以下主要功能能力：活组织切片中Ca 2+瞬变和荧光标记物的共聚焦成像; Ca 2+瞬变的比率Fura-2成像;在离子通道活性的伴随膜片钳记录期间Ca 2+成像的高时间和空间分辨率;以及笼状化合物的UV闪光光解。所有主要用户都具有丰富的活细胞钙成像、电生理学和/或共聚焦显微镜经验。总的来说，我们已经发表了一百多篇论文，使用一种或多种这些技术，其中包括15篇论文，同时使用功能共聚焦成像和电生理记录活细胞。激光共聚焦显微镜的一个固有问题是非常强烈的激发光，导致大量的染料分解（光漂白）在活细胞成像。所提出的奥林巴斯BX-DSU活细胞盘扫描成像系统通过优化信号检测克服了这个问题。它采用旋转圆盘共焦技术和新一代超冷EM-CCD相机，生成具有高时间和空间分辨率的图像。宽光谱光源产生的光漂白比激光少得多。此外，该系统还增加了一个UV闪光灯照明器，用于光活化（笼状）化合物的光解，完整集成了用于伴随电生理记录的膜片钳仪器，以及组织切片工作的直立支架。其结果是一个非常具有成本效益和灵活的解决方案，响应所有描述的主要用户的需求。肯塔基州大学或附近没有其他仪器可以提供这种特征。联合国大学将提供适当的空间，为有经验的技术人员提供工资支持，并为系统提供维护。该系统将由扫描磁盘技术和电生理学专家在肯塔基州大学的一个大型完善的成像设施内建立。该系统将允许主要用户继续实现新的发现，以更好地理解和治疗慢性疼痛，肥胖，听觉障碍，嗅觉障碍，肺部疾病，脑损伤的后果和心血管疾病。

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.