Regensburg 2007 – scientific programme

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BP: Fachverband Biologische Physik

BP 26: Poster Session II

BP 26.11: Poster

Thursday, March 29, 2007, 17:00–19:30, Poster B

EMCCD-based spatially and spectrally resolved fluorescence correlation spectroscopy — •Markus Burkhardt, Jonas Ries, and Petra Schwille — Biophysics/ BIOTEC/ TU Dresden, Tatzberg 47-51, 01307 Dresden, Germany

Fluorescence Correlation Spectroscopy (FCS) is based on time dependent fluorescence intensity fluctuations of labeled biomolecules as they enter and leave a diffraction-limited optical detection volume. From simple autocorrelation analysis, concentrations, diffusion and binding coefficients are easily obtained.

Spectral and spatial cross-correlation enhances the sensitivity and accuracy of FCS measurements to determine exact concentrations of interacting partners and to obtain absolute diffusion coefficients, respectively. Both tasks can conveniently be performed employing an electron multiplying CCD camera for detection of the fluorescence signal. Applying different readout modes of the CCD enhances the data acquisition speed and therefore the time resolution needed for FCS.

We demonstrate spatial cross correlation with different focal geometries as well as spectrally resolved FCS using EMCCD-based detection.