Fig. 3. Storage of neutral lipids. Parasites grown in the presence of the fluorescent fatty acid C4C9 were added either to poly-L-lysine-coated coverslips (extracellular) or coverslip-grown host cells. After 1 hour (extracellular and short infection) or 6 hours (6h infection), coverslips were fixed and processed for immunocytochemistry using antibodies directed against MIC4, ROP2 or GRA2. Primary antibodies were detected by subsequent incubation with Cy5-conjugated anti-mouse or anti-rabbit antibodies. Additional coverslips were incubated with the lipophilic dye Nile Red or the nucleic acid stain TOPRO3. Images were acquired by confocal microscopy. Single, 0.4 µm thick optical sections are shown. The immunochemical staining of parasite organelles appears blue, whereas C4C9 is depicted in green. The two channels merged onto the brightfield image are shown to facilitate the comparison of labeling patterns. Bar, 5 µm.

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