Pixel-wise rate coefficients of fluorescence loss kinetics of eGFP. Using the parameter maps of the FLIP analysis for the cell in Figure 4, rate coefficients were calculated for every pixel position, as outlined in Additional file. A, the resulting 32-bit image stack was color-coded using a FIRE-LUT, and selected frames of the rate coefficients were plotted (blue and yellow-white indicate low and high rate coefficients, respectively). Areas with accelerating speed of FL turn from blue to yellow-white over time. A’ few regions are highlighted with boxes (numbered ‘1’ to ‘5’) for further analysis. Boxes 1 and 2 were placed in the nucleus, while boxes 3 to 5 were in the cytoplasm (inset shows a zoom for the area containing boxes 3 and 4 close to the right edge of the cell). B, rate coefficients as function of time plotted for box 1 to 4. C, x,t-view of the time evolution of the rate coefficients in box 5. See text for further explanations.