CORE--CONFOCAL MICROSCOPY

最新推出的尼康RCM 8000扫描仪之一 阿尔伯特·爱因斯坦肯尼迪中心的共焦显微镜 医学院，该计划项目中的所有私家侦探都有 他们的实验室。该指令超越了传统的共焦指令 显微镜，以其空间分辨率著称，实时完全 以每帧30帧的速率捕获的场比度量测量值 其次，对于较小的领域，甚至更快，同时保持 真正的共聚焦。因此，诸如细胞内和 细胞内钙波可以在时间和空间上被监测 活细胞的出现，通过当代的应用 钙指示剂(如吲哚和氟3)。钙离子波的分析 将在星形胶质细胞中进行(项目2)，系列重建 荧光黄染料填充损伤和正常神经的雪旺细胞 野生型和Cx32 KO动物(L计划)也将利用 仪器的快速扫描能力。此外， 显微镜将用于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 项目。这台显微镜的出色空间分辨率将是 不仅对生成缝隙连接的重建图像有价值 在这个项目中的分布，但将使我们能够确定 可能存在缺陷的突变型连接蛋白的亚细胞分布 向质膜运输(项目3)。

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).