Figure 1. Fluorescence decay after... | Rockefeller University Press

Figure 1.

$Figure 1. Fluorescence decay after single photoactivation (single FDAP) of Dp-actin. (A) Fluorescence image of Dp-actin in an MCF-7 cell. (B) Fluorescence image of a 4.8 x 3-µm region (A, white box) after photoactivation of the whole cell to the maximum level. (C) Image after photobleaching. (D) Time-lapse images after photoactivation of a 1.8-µm-diameter area (solid circle) for 3 ms. Images were acquired every 30 ms for 270 ms. (E) Time course of Dp-actin fluorescence decay after single photoactivation. The relative intensity in a 3-µm-diameter area (D, dotted circle) was plotted (white diamond), after correcting (Fig. S1) and normalizing to the maximum intensity in the same area, measured as in B. The diffusion coefficient and mobile fraction of Dp-actin were determined by fitting the time curve of fluorescence decay (black line). The black circle indicates the normalized fluorescence intensity of immobilized Dp after single photoactivation in fixed cells. Data are means ± SEM of 22 cells (Dp-actin) and 25 cells (Dp) from triplicate experiments. Bars, 20 µm.$

Fluorescence decay after single photoactivation (single FDAP) of Dp-actin. (A) Fluorescence image of Dp-actin in an MCF-7 cell. (B) Fluorescence image of a 4.8 x 3-µm region (A, white box) after photoactivation of the whole cell to the maximum level. (C) Image after photobleaching. (D) Time-lapse images after photoactivation of a 1.8-µm-diameter area (solid circle) for 3 ms. Images were acquired every 30 ms for 270 ms. (E) Time course of Dp-actin fluorescence decay after single photoactivation. The relative intensity in a 3-µm-diameter area (D, dotted circle) was plotted (white diamond), after correcting (Fig. S1) and normalizing to the maximum intensity in the same area, measured as in B. The diffusion coefficient and mobile fraction of Dp-actin were determined by fitting the time curve of fluorescence decay (black line). The black circle indicates the normalized fluorescence intensity of immobilized Dp after single photoactivation in fixed cells. Data are means ± SEM of 22 cells (Dp-actin) and 25 cells (Dp) from triplicate experiments. Bars, 20 µm.