Figure 10.

The fluorescence from Galtase-GFP in a nocodazole-treated (1 μM) treated blastomere was bleached using a FLIP protocol. The image quality is poorer than that shown in Figure 8, because lower illumination levels were used to minimize bleaching. Photobleaching using a FLIP protocol was begun 52 min after nocodazole was added. The protocol was essentially the same as used in Figure 3, except that the wait period was 20 s instead of 10 s, and the relative zoom of bleached to imaged was 6 instead of 5. The duration of each cycle was 30 s, and the total elapsed time for the 10 cycles was 5 min. Fluorescence was diminished throughout the cell, providing strong evidence that Galtase-GFP is in the ER. The size of the field imaged was 59 × 39 μm, and the bleached area was 12 × 7 μm and is indicated by the rectangle. Bar, 10 μm.