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

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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.