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Measured diffusion coefficient of Atto655 maleimide in aqueous solution at 25°C as a function of excitation power. Diffusion coefficients measured using pmFCS (squares) and 2fFCS (diamonds) are virtually independent of the excitation power up to 200 µW. The solid line is the average of these values. The results of the single-focus FCS (circles) and the second order polynomial fits (dashed line) are shown in comparison.
