Fig. 1.
3PEA concept. (A) A CLSM acquires and bleaches images via a pixel-by-pixel scanning process in a sequential manner. This is not considered in the conventional FRAP analysis. In contrast, 3PEA explicitly calculates and fits the spatiotemporal intensity distribution generated by the sequential bleach process. (B) Time evolution of the intensity profiles for particles with different diffusion coefficients during and after the bleach process. The profiles were calculated for a 128 × 128 image size, a pixel time τp of 10 μs, and a line time τl of 1 ms. The bleach spot is depicted in white.