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FRAP, fluorescence recovery after photobleaching; GFP, green fluorescent protein; msFCCS, multi-scale fluorescence cross-correlation spectroscopy; RFP, red fluorescent protein.
Data were either obtained by msFCCS or FRAP with radial profile analysis. The results for TetraSpeck beads in vitro are in excellent agreement with the value D=(4.4±0.7) μm2 s−1 previously determined by dual-focus FCS and dynamic light scattering69. According to the measured diffusion coefficient of QDots, a hydrodynamic radius of rH=(7.8±0.3) nm was calculated that matches the specification value of (8.8±1.3) nm given by the manufacturer (Invitrogen). msFCCS results in living cells agreed very well with FRAP experiments on the same length scale.
*Diffusion constants obtained by auto-correlation analysis are reported, since results were independent on time and length scale.
†Diffusion constants for msFCCS analysis with an effective distance of deff=1.2 μm.
‡FRAP results for a bleach circle radius of wb≈1.3 μm.
§An immobile fraction of (1±1) % was found on the minute time scale.
∥An immobile fraction of (6±1) % was found on the minute time scale.