CORE--CONFOCAL MICROSCOPY

核心——共焦显微镜

• 批准号: 6345004
• 负责人: David C Spray
• 金额: $ 24.81万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6345004
• 关键词: astrocytes; bioimaging /biomedical imaging; biomedical facility; calcium flux; confocal scanning microscopy; gap junctions; membrane channels

One of the first of the recently-introduced Nikon RCM 8000 scanning confocal microscropes is housed at the Kennedy Center of Albert Einstein College of Medicine, where all of the P.I.s on this Program Project have their laboratories. This instruction moves beyond conventional confocal microscopy, noted for its spatial resolution capacity, to real time full field ratio metric measurements captured at a rate of 30 frames per second, or even faster for smaller parts of the field, while maintaining true confocality. Hence processes such as intracellular and intercellular Ca2+ waves can be monitored temporally and spatially in living cells as they occur, by the application of the current generation of calcium indicators (e.g. Indol and Fluo3). Analysis of Ca2+ waves will be undertaken in astrocytes (Project 2), serial reconstructions of Lucifer yellow dye fills in Schwann cells from injured and normal nerves of wildtype and Cx32 KO animals (Project l) will also take advantage of the rapid scanning capability of the instrument. In addition, the microscope will be used for immunocytological detection of connexin distribution in cells, or in double label experiments, in all of the projects. The excellent spatial resolution of this microscope will be of value not only for generating reconstructed images of gap junction distribution in this projects, but will enable us to determine subcellular distribution of mutant connexins that may be defective in transport to the plasma membrane (Project 3).

最新推出的尼康RCM 8000扫描仪之一 共焦显微镜被安置在阿尔伯特·爱因斯坦的肯尼迪中心 医学院，在那里，所有的PI在这个项目上有 他们的实验室。该指令超越了传统的共焦 显微镜，指出其空间分辨率能力，以真实的时间充分 以每帧30帧的速率捕获的场比率度量测量 第二，甚至更快的领域的较小部分，同时保持 真正的共焦因此，诸如细胞内和 细胞间Ca ~（2+）波可以在时间和空间上进行监测， 活细胞，因为他们发生，通过应用当前的一代， 钙指示剂（例如Indol和Fluo 3）。Ca 2+波分析 将在星形胶质细胞中进行（项目2）， 荧光黄染料填充损伤和正常神经的雪旺细胞 野生型和Cx 32 KO动物（项目1）也将利用 仪器的快速扫描能力。此外该 将使用显微镜进行连接蛋白的免疫细胞学检测 在细胞中的分布，或在双标记实验中，在所有的 项目这台显微镜出色的空间分辨率将是 的价值，不仅用于生成间隙连接的重建图像， 分布在这个项目中，但将使我们能够确定 突变连接蛋白的亚细胞分布可能是缺陷的， 运输到质膜（项目3）。