Microglia Phenotyping by Flow Cytometry

Flow cytometry was used to detect the expression of CD16/32 and CD206 in microglia cells as previously described [31 (link)]. In brief, 100 μl microglia cell suspension (1 × 10⁶ cells) was aliquoted into 1 ml Eppendorf tubes. The anti-mouse CD16/32 PE (0.125 μg, Biolegend, CA, USA) and anti-mouse CD206 (MMr) FITC (0.125 μg, Biolegend, CA, USA) were added into the EP tubers after 0.5 h of incubation at 4 °C, The microglia cells were washed with PBS twice and resuspended in 300 μl 1 × PBS solution. Analyses were performed on Becton-Dickinson FACS Calibur (Becton Dickinson, Bedford, MA). Mean fluorescence intensity was measured.