Fig. 2

Microscopic observation of phagocytosis of late GCT by immortalized mouse C57Bl/6 bone marrow-derived macrophages. a BMDM were incubated with GCT for 2 h, detached from the plate, spotted on smears by cytospin centrifugation, fixed in absolute methanol, and stained with Giemsa. The arrows indicate GCT inside macrophages. Pictures, from one representative experiment out of three, were taken at ×1000 magnification. b GCT were incubated with 3 µM CellTracker™RedCMTPX Dye for 30 min to stain cytoplasm (red in the picture). BMDM were incubated for 2 h with GCT and fixed with formalin 4 %. BMDM membranes were stained with 10 µg/ml WGA-FITC (Wheat germ agglutinin, DBA Italia) (green in the picture). BMDM nuclei were stained by the DAPI present in the mounting medium (Fluoroshield) (blue in the picture). c Z-stack images were taken on an inverted microscope equipped with a confocal spectral imaging system (Olympus Fluoview 1000, Tokyo, Japan) 600× magnification. Scale bar, 10 µm