Abstract
A novel, high signal-to-noise strategy is presented for phase-selective, wavevector- and polarization-resolved fluorescence fluctuation spectroscopy. The approach is based on molecular Fourier imaging correlation spectroscopy (MFICS) and combines polarization- and intensity-modulated photoexcitation with phase-synchronous detection to simultaneously monitor centre-of-mass and slow fluorescence anisotropy fluctuations from a relatively large number (∼106) of fluorescent molecules. The method is demonstrated using DsRed, a tetrameric complex of fluorescent protein subunits, to unambiguously separate signal contributions due to slow optical conformational fluctuations from translational diffusion.
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