Fig. 2. Photobleaching and recovery of ensconsin chimera in untreated cells. (A) Time-lapse micrographs of the bleached area before (pre-bleach) at the time of bleaching (laser spot) and at intervals during the recovery. Elapsed time, in seconds, is shown in each micrograph. (B) Corrected fluorescence (total pixel intensity along a segment of a bleached MT) was plotted against time (in seconds) before and after each of two bleach events. The time of each bleach is marked with an arrow. Fluorescence shown has been corrected, using an unbleached segment of an adjacent MT, for decay caused by photobleaching during time-lapse imaging and cellular background has been subtracted. (C) Determination of kOFF. As described in Materials and Methods, the logarithm of the fraction of fluorescence recovered was plotted against time and the slope, kOFF, was determined by linear regression.
