OLYMPUS DSU CONFOCAL SYSTEM: ZEBRAFISH:POLYCYSTIC KIDNEY DISEASE

奥林巴斯 DSU 共焦系统：斑马鱼：多囊肾病

• 批准号: 7335305
• 负责人: Lynn COOLEY
• 金额: $ 1.33万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-15 至 2007-03-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7335305
• 关键词: Caenorhabditis elegans; Drosophilidae; antibody; cell migration; charge coupled device camera; confocal scanning microscopy; fasting; fluorescence; gene mutation; genetics; housing; kidney; lasers; ligands; light intensity; lighting; mass information media; microscopy; molecular /cellular imaging; mutant; phosphatidylinositols; polycystic kidney; proteins; receptor; tissue /cell culture; tissues; zebrafish

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): The ability to obtain high-resolution, live-cell images and movies is essential for understanding biological processes and how they are perturbed by disease or mutation. This proposal is for the purchase of an Olympus Disc Scan Unit (DSU) confocal microscope system. Unlike laser-scanning confocal microscopy, the Olympus DSU system uses a white light source, thus dramatically reducing the initial cost and maintenance of the system. To compensate for the lower light intensity available from white-light sources, the Olympus DSU system uses a highly sensitive cooled charged-couple device (CCD) camera to capture fluorescence signals. Out-of-focus light is eliminated by the spinning disc with precisely defined slits that create virtual pin-hole illumination. Fast image acquistion (up to 15 frames/sec) and lowered phototoxicity from spinning disc illumination combine to allow high-quality imaging of both fixed and live cells. The Olympus DSU system will be housed in the Department of Genetics and used by investigators for a variety of projects funded by NIH R01 grants. These include imaging of hundreds of GFP-fusion proteins in Drosophila tissues (Cooley), antibody localization in mouse and C. elegans germline cells (Reinke, Sweasy), cell migration in C. elegans (Stern), localization of mutant forms of PDGF receptor in cells treated with ligands (DiMaio), phosphoinositide signaling in cultured cells (H. Sun), and live imaging of developing zebrafish kidneys (Z. Sun). The Olympus DSU system will replace two out-dated, non-motorized fluorescence microscopes that can no longer produce data essential for these projects. It will also provide a much-needed alternative to expensive laser-scanning confocal imaging, especially in cases where the intensity of fluorescence signals is not limiting. Furthermore, the system will allow ready access to time-lapse microscopy, again with a reduced cost compared to laser-based systems.

该子项目是利用由NIH/NCRR资助的共享仪器赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。列出的机构是用于资助的，不一定是研究者的机构。描述（由申请人提供）：获得高分辨率活细胞图像和电影的能力对于理解生物过程以及它们如何受到疾病或突变的干扰至关重要。该提案是为购买奥林巴斯光盘扫描单元（DSU）共聚焦显微镜系统。与激光扫描共聚焦显微镜不同，Olympus DSU系统使用白色光源，从而大大降低了系统的初始成本和维护。为了补偿白光源的低光强，Olympus DSU系统使用高灵敏度的冷却电荷耦合器件（CCD）相机来捕获荧光信号。旋转的圆盘带有精确定义的狭缝，可创建虚拟针孔照明，从而消除失焦光线。快速图像采集（高达15帧/秒）和旋转圆盘照明的低光毒性结合联合收割机，可对固定细胞和活细胞进行高质量成像。奥林巴斯DSU系统将被安置在遗传学系，并由研究人员用于NIH R 01资助的各种项目。这些包括果蝇组织中数百种GFP融合蛋白的成像，小鼠和C. elegans生殖系细胞（Reinke，Sweasy）、C. elegans（Stern），用配体处理的细胞中PDGF受体的突变形式的定位（DiMaio），培养细胞中的磷酸肌醇信号传导（H. Sun），以及发育中的斑马鱼肾脏的实时成像（Z。Sun）。奥林巴斯DSU系统将取代两台过时的非电动荧光显微镜，这些显微镜不再能产生这些项目所需的数据。它还将为昂贵的激光扫描共聚焦成像提供急需的替代方案，特别是在荧光信号强度不受限制的情况下。此外，该系统将允许随时访问延时显微镜，与基于激光的系统相比，成本也降低了。