Filtered FCS and species cross correlation function

An extended analysis method of time, polarization and color resolved fluorescence correlation spectroscopy (filtered FCS) is introduced. It uses multiparameter fluorescence detection (MFD) [1-3] to separate pure fluorescence signal of multiple species and scatter contributions. This method allows monitoring of simultaneous and independent diffusion of several molecular species in one sample and makes possible accurate and quantitative analyses of fractions. The proposed method is simple to implement experimentally, because it requires only single wavelength excitation and MFD widely used in single molecule experiments. In comparison to recently introduced fluorescence lifetime correlation spectroscopy (FLCS) [4-7] this method is able to distinguish species when they have very close or even the same fluorescence lifetime using just differences in time resolved fluorescence anisotropy.

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