Regensburg 2004 – wissenschaftliches Programm
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SYLS: Life Sciences on the Nanometer Scale - Physics Meets Biology
SYLS 3: Symposium "Life Sciences on the Nanometer Scale - Physics Meets Biology"
SYLS 3.42: Poster
Mittwoch, 10. März 2004, 16:00–18:30, B
Diffusion in live cells: A combined optical sectioning and single particle tracking approach — •Ralf Bausinger1, Christoph Bräuchle1, Sabine Boeckle2, Katharina von Gersdorff2, Ernst Wagner2, and Andreas Zumbusch1 — 1Department Chemie - LMU München - Butenandtstr. 5-13 - D-81377 München — 2Department Pharmazie - LMU München - Butenandtstr. 5-13 - D-81377 München
Single particle tracking methods with sensitivities down to the single molecule detection limit have become important tools for the study of diffusion processes. For their application to studies of trafficking in live cells, the trajectories have to be analyzed in the context of the complex cellular structures. In order to visualize these structures in three dimensions, confocal fluorescence microscopy is commonly applied because of its high axial resolution. This raster scanning method however contradicts the simultaneous usage of wide-field microscopy usually employed for particle tracking. Classical wide-field microscopy in turn does not offer sufficient axial resolution in order to reconstruct the three dimensional image of the cell.
For this reason we use structured wide-field illumination in order to achieve high axial resolution. Like this, fast switching between high resolution cellular imaging and tracking with single molecule sensitivity becomes possible. The application of this setup to studies of artificial virus diffusion in live cells is demonstrated.