Phagocytosis Assay for Microglial Cells

Primary microglia isolated from WT adult mice were plated on poly-D-lysine-coated glass coverslips. Cells were treated with latex beads-rabbit IgG-FITC complex (Cayman Chemical, Ann Arbor, MI) at a dilution of 1:200 for 30 min. For LPS-stimulated microglia, cells were stimulated with 1 μg/mL LPS for 1 h under serum-free condition, followed by adding IgG-FITC complex and monitoring phagocytosis by immunostaining [14 (link), 26 (link)]. Briefly, cells were washed at least three times with a warm (37 °C) serum-free medium and then fixed with 4 % paraformaldehyde. Fixed cells were processed for Iba1 immunostaining using the procedure described above. Imaging was performed under Olympus BX41 fluorescence microscope.