Disruption of lipid rafts inhibits <i>C</i>. <i>albicans</i> phagocytosis by human monocytes.

<p><b>A.</b> Phagocytosis inhibition assessed by flow cytometry. Human monocytes were pretreated with MBCD, DAmB or left untreated (none), and then incubated with HK FITC-conjugated <i>C. albicans</i> or <i>Staphylococcus aureus</i> (<i>St. aureus</i>). The data highlight that drugs specifically impair fungal phagocytosis. Markers were set up to exclude background staining of cells incubated with the microorganisms at 0°C. Numbers indicate the percentage of monocytes with ingested microorganisms. Data shown are from one experiment representative of four. <b>B.</b> Phagocytosis analysis by confocal microscopy. Phagocytosis was performed as in A. Samples were acquired in 3D stacks by DIC and three-color fluorescent images. The triple staining allows to clearly establish if a FITC-conjugated <i>C</i>. <i>albicans</i> (green) was internalized by a monocyte labelled by anti-CD14 (red). DNA was counterstained with DAPI (blue). The first column (DIC overlay) shows the combined signals from the DIC and confocal fluorescent channels of a single xy slice. In the second column (3D rendering), the 3D reconstructions of the 22 slices highlighting the reciprocal spatial information of fungal and human cells are displayed. The third column (orthogonal views) shows the xy, xz and yz single planes at the indicated orthogonal xyz axes (white lines). Scale bar for all panels is 3 μ.