Dresden 2014 – scientific programme

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BP: Fachverband Biologische Physik

BP 12: Posters: Imaging

BP 12.1: Poster

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

Homo-Fret to Investigate the Oligomeric State of Proteins — •Franz-Josef Schmitt, Matthias Broser, Cornelia Junghans, and Thomas Friedrich — Institute of Chemistry, Biophysical Chemistry, TU Berlin, Straße des 17. Juni 135, D-10623 Berlin, Germany

The combination of various microscopic techniques allows to address complex problems in biophysics. We present a multi-parameter setup based on to a Nikon TI Eclipse microscope that combines time resolved fluorescence microscopy and simultaneous anisotropy microscopy by splitting the detector optically into two independent areas. All microscopic techniques (confocal, wide field, total internal reflection) can be done with a high photon throughput up to 2 Mio. photons/sec. with spectral and polarization resolution. Förster Resonance Energy Transfer (FRET) between identical fluorophores without the discrimination between donor and acceptor (Homo-FRET) allows the determination of the aggregation state of identical chromophors. Microscopic measurements of time resolved Homo-FRET were used to determine the aggregation state of a model system of fluorescence proteins (FKBP-GFP constructs) that can dimerize or form larger aggregates (up to pentamers) by adding a specific membrane-permeable agent. The technique has great application potential for the observation of the repair cycle of the D1 protein in the photosystem II.