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Fig. 2 | BMC Immunology

Fig. 2

From: TIRF imaging of Fc gamma receptor microclusters dynamics and signaling on macrophages during frustrated phagocytosis

Fig. 2

IgG-coated supported lipid bilayer permits selective activation FcγR on macrophages. a Cartoon of the supported lipid bilayer (SLB) presentation of IgG via its interaction with Biotin-PE. Bone marrow derived macrophages (BMM) were dropped onto this surface and the plasma membrane in contact with the glass was imaged by total internal reflection fluorescence (TIRF) microscopy. Upon contact with IgG-coated SLB containing 1 % Biotin-PE, BMM labeled with DiI spread and form multiple dynamic pseudopodia, analogous to phagocytosis (phagocytic pseudopod extension). c BMM did not engage SLB lacking IgG and simply rolled along the surface. d-f Alexa Fluor 594-IgG, allowed imaging of the dynamics of FcγR-IgG complexes during the initial interaction and spreading phase (d) during which they formed microclusters that moved with the extending pseudopod. e After about 3–5 min (montage starts about 3 min after adding BMM to the SLB), FcγR-IgG microclusters would traffic toward the center of the cell, where they would accumulate. f Magnified montage showing the early stages of cell spreading, FcγR-IgG were observed to decorate the advancing pseudopod edge, whereupon they would continue to cluster and eventually begin a retrograde motion toward the center of the cell. Scale bar 10 μm for (b-c) and 5 μm for (d-f)

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