Regensburg 2013 – scientific programme

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BP: Fachverband Biologische Physik

BP 8: Posters: Proteins

BP 8.30: Poster

Monday, March 11, 2013, 17:30–19:30, Poster B2

Pitfalls and artifacts in two-focus fluorescence fluctuation spectroscopy — •Andreas Veres and Matthias Weiss — Experimental Physics I, University of Bayreuth, Germany

Fluorescence fluctuation spectroscopy is a powerful technique to quantify intracellular transport on the almost single-molecule scale. Going beyond a single observation volume, temporally cross-correlating the fluorescence from two separated foci can be exploited to obtain large-scale transport coefficients, and to detect transport barriers or directed flow. Using mean-field theory and simulations in combination with experiments on a laser scanning microscope (LSM), we have investigated which artifacts and pitfalls may hamper this promising approach. We find that the unavoidable bleaching of fluorophores only has a minor influence on the results, whereas the typically poor statistics in commercial LSMs severely limits the applicability of the approach to in vitro systems.