Fig. 4

The first four images a-d are the original FLIP images of the CHO cells expressing GFP-Q73 in the cytoplasm and nucleus. It is produced in a temperature controlled (35±1°C) environment on a Zeiss LSM 510 confocal microscope using the 488nm line of an Argon laser. The black circle on the image a shows the 25-pixel wide bleaching area and a scalebar which is 5 μm. The pixel size is here 0.0461847 μm given a width of the bleach spot of ca. 1.15 μm. The leftmost FLIP image a is taken before bleaching, the next image b is taken after it has been bleached 10 times, i.e., time t=28 s. The third FLIP image c is the 20’th FLIP image in the sequence (time t=56 s) and the last d is at time t=109.2 s which correspond to FLIP fame 39. The second row e-h shows the corresponding goal function. The third row i-l shows the simulation results, all at times corresponding to the displayed FLIP images