Figure 2.

CRY2 recruitment as a function of illumination characteristics. (a) Differential TIRF image of pmCRY2 3s after the illumination with a local pulse of blue light. (White line) Contour of the cell. (Side curves) pmCRY2 intensity (green) and Gaussian fit (red) along a line across the activation area (red circle). Scale bar = 10 μm. (b) Quantification of the mean intensity in the activation area (red circle, a) divided by its mean value before activation as a function of time for activating pulses of different duration in a single cell. The exposure times were chosen equal to 3, 9, 18, 50, 100, and 200 ms. Images are taken every 5 s. The temporal decay (∼80 s) is governed by lateral diffusion because the diffusion time is smaller than the complex dissociation (185 s) for such point activations. (c–f) Mean values (blue dots) and standard deviation (blue-shaded regions) of pmCRY initial Gaussian distribution after a single pulse of activation. The average width σ (c and d), and total integrated amount (e and f) calculated from the integral of the Gaussian 2πσ_xσ_yA are plotted as a function of laser power (c and e) and exposure time (d and f). In (d) and (f) the laser power is fixed at 5.5 μW and the pulse durations are 3, 9, 18, 50, 100, and 200 ms (N = 15 cells). In (c) and (e) the exposure is fixed at 50 ms and the laser power is set to 4.5, 5.5, 9, 21, and 36 μW (N = 25 cells). To see this figure in color, go online.