Confocal imaging of intracellular protein dynamics. (360G-Wellcome-078020_Z_05_Z)

For many processes in cell biology, the key proteins involved have now been identified. Yet we remain far from understanding how these proteins function because information on how they respond to different cellular cues and interact on a temporal and spatial basis in vivo is lacking. We propose to use state-of-the-art imaging technology to tackle this problem through experimental observation of the localization, interaction and dynamics of proteins in living cells. We will specifically examine the assembly and organization of subcellular macromolecular machines including the centrosome, focal adhesion complexes, signalling pathway scaffolds, spliceosomes and the nuclear envelope. This will complement and enhance Wellcome Trust funded projects in each of these areas. To make a real impact, we request funding to purchase a Leica AOBS laser scanning confocal fluorescence microscope with appropriate lasers and software modules that will enable us to perform quantitative, multiwavelength, time-lapse imaging, FRET and photobleaching studies. The requested equipment will be incorporated into the epifluorescence microscope facility set up in Leicester with funding from the Wellcome Trust in 2001 and managed by the grant-funded technician. This will provide us with an entirely new range of imaging technologies enabling us to remain internationally competitive in our research areas.