Phagocyte and budded yeast. Amoebae (D. discoideum) were plated with yeast (S. cerevisiae) in a glass bottom chamber and viewed by confocal microscopy. Fluorescence is superimposed on grey scale brightfield images. The amoebae were expressing LimEdelta-GFP to label actin filaments. The interaction between an amoeba and two budded yeast is shown. When a phagocytic cup reaches the constriction at the neck of the first budded yeast, actin accumulates there as the amoeba attempts to seal the cup. When this attempt fails, the amoeba releases the yeast. Another budded yeast is then encountered and similarly attacked, also unsuccessfully. Finally, the amoeba returns to the first yeast and this time succeeds in separating the yeast mother and bud, allowing one portion to be ingested. The time series was captured in a single focal plane at 3.94-second intervals using a Zeiss LSM 510 laser scanning confocal microscope equipped with a 63x 1.4 NA DIC objective lens. Pixel size 0.08 x 0.08 microns. The movie plays at 30x actual speed. Correspond to Movie 1 in BMC Biol. 2010 Dec 29;8:154.