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BP: Fachverband Biologische Physik

BP 12: Posters: New Technologies

BP 12.15: Poster

Montag, 14. März 2011, 17:15–20:00, P3

Detection of conformational states of proteins in C. elegans in vivo by confocal fluorescence anisotropy — •Volker Christoph Henschel¹, Alessandro Esposito², Eugenia Butekevich¹, Christoph Friedrich Schmidt¹, Fred Sylvester Wouters³, and Dieter Robert Klopfenstein¹ — ¹Drittes Physikalisches Institut, Biophysics, Georg-August-University Göttingen, Göttingen, Germany — ²MRC Cancer Cell Unit, Hutchison/MRC Research Centre, Cambridge, UK — ³Laboratory of Cellular and Molecular Systems, Department of Neuro- and Sensory Physiology, Georg-August-University, Göttingen, Germany

The study of protein-protein interactions in vivo is often hindered by the limited acquisition speed of typical instrumentation used, for instance, for lifetime imaging microscopy. Anisotropy polarization is altered by the occurrence of Foerster Resonance Energy Transfer (FRET) and anisotropy imaging was shown to be comparatively fast and simple to implement. Here, we present the adaptation of a spinning disc confocal microscope for fluorescence anisotropy imaging that allowed to achieve in vivo imaging at high spatial and temporal resolution. We demonstrate the capabilities of this system and in-house developed analysis software by imaging living Caenorhabditis elegans expressing constitutive dimeric and monomeric proteins that were tagged with GFP.