Transport pathways of single molecules and nanoparticles in the nucleus of the living cell

在过去的项目期间，我们实现了有史以来第一次在细胞核内观察到单个可移动的蛋白质分子。新的实验能力现在将致力于研究小核核糖核蛋白(SnRNP)颗粒的核内运输，这些颗粒在核质中表现出复杂的、可能是动态的定位模式，因为它们与几个不同的核亚结构有关，如染色质周围纤维、染色质间颗粒斑点和卷曲小体。通过对野生型和突变型U SnRNP颗粒的高速和高分辨率成像，我们想要检查它们的迁移率，并可能与被引用的通透性细胞核内的结构有关。为了实现这一目标，应同时通过双色荧光对第二分子(参考)组分的额外可视化以及对样品的轴向延伸区域的高速成像来扩展仪器容量，以跟踪完整的三维轨迹。

In the past project period we realized the first observations of single mobile protein molecules inside cell nuclei ever. The new experimental capabilities shall now be devoted to studying the intranuclear transport of small nuclear ribonucleoprotein (snRNP) particles that display a complex, probably dynamical localization pattern in the nucleoplasm because of their association with several distinct nuclear substructures such as perichromatin fibrils, interchromatin granule speckles and coiled bodies. By high-speed and high-resolution imaging of wildtype and mutant U snRNP particles we want to examine their mobility, and possibly association with the cited structures inside nuclei of permeabilized cells. To reach this goal the instrumental capacities shall simultaneously be extended by additional visualization of a second molecular (reference) component by dual-color fluorescence, and high-speed imaging of axially extended regions of the specimen to follow complete three-dimensional trajectories.