The most abundant immune cells in Drosophila are macrophage-like plasmatocytes that fulfill central roles in morphogenesis, immune and tissue damage response. The various genetic tools available in Drosophila together with high-resolution and live-imaging microscopy techniques make Drosophila macrophages an excellent model system that combines many advantages of cultured cells with in vivo genetics. Here, we describe the isolation and staining of macrophages from larvae for ex vivo structured illumination microscopy (SIM), the preparation of white prepupae for in vivo 2D random cell migration analysis, and the preparation of pupae (18 h after puparium formation, APF) for in vivo 3D directed cell migration analysis upon wounding using spinning disk microscopy.
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