Fig. 6. PtdIns(3,4,5)P₃ generation at individual phagocytic cups. (a) A micropipette was used to present two iC3b-opsonised zymosan particles, (labelled Z1 and Z2) to a neutrophilic HL60 cell shown in the phase-contrast image and the simultaneously acquired confocal image of the distribution of GFP-Akt-PH (green). The fluorescent images from the zone indicated by the rectangle are shown below at 10 second intervals, with the location of particles Z1 and Z2 indicated on the first image. The complete data set is shown as a movie (Movie 4 in supplementary material) and the time course of GFP-Akt-PH accumulation in the membranes at these two locations is shown in b. Example of the distribution of GFP-Akt-PH in two phagosomes sequentially formed and touching (c) and simultaneously forming at opposite poles of the cell (d). In c and d the images show GFP-Akt-PH distribution at 20 second intervals. Bars, 2 µm.