Dresden 2014 – scientific programme

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BP: Fachverband Biologische Physik

BP 12: Posters: Imaging

BP 12.2: Poster

Tuesday, April 1, 2014, 09:30–12:30, P1

Diffusion in Living Cells Determined by Multiparameter Imaging — •Franz-Josef Schmitt, Cornelia Junghans, Matthias Sturm, Marvin Schlischka, and Thomas Friedrich — Institute of Chemistry, Biophysical Chemistry, TU Berlin, Straße des 17. Juni 135, D-10623 Berlin, Germany

Multiparameter microscopy is close to becoming a standard technique in live cell imaging. The synergistic interplay between highly resolved fluorescence microscopy and photoswitchable fluorescence proteins lead to a brilliant new methodology that broke the diffraction barrier of optical microscopy. We show how the photoswitchable genetically expressed Dreiklang Protein is used to determine the local diffusion coefficients in the cell cytoplasm and the cell nucleus as well as the kinetics of the Dreiklang protein diffusion across membranes. The technique shows preferred pathways of proteins across the cell nucleus which are not visible by constantly fluorescing labels due to the strong overlap of the emission of single molecules that lead to homogeneous fluorescence patterns. The dynamics of selectively marked mitochondria is tracked and the mean square displacement is calculated showing regimes of hampered diffusion, free diffusion and active transport.