Immunofluorescence Imaging of Microglia Markers

Sections of the prefrontal cortex (PFC) were incubated with primary antibodies overnight and then secondary antibodies in the dark for 1 h. The antibodies used for IF staining were as follows: anti-IBA-1 (1:200, GB12105, Servicebio), anti-CD68 (1:100, GB113109, Servicebio), anti-CR3 (1:500, GB11058, Servicebio), CY3 goat anti-mouse secondary antibody (1:300, GB21303, Servicebio), and 448 AffiniPure Fab Fragment goat anti-rabbit secondary antibody (1:500, GB25303, Servicebio). The brain slices in our study underwent a blocking step using 10% goat normal serum [29 (link)]. Samples were counterstained with 4,6-diamidino-2-phenylindole (DAPI) for 10 min and then visualized using a fluorescence microscope (Nikon Eclipse C1, Japan).