Multimodal Immunolabeling of Brain Slices

Immunofluorescence was performed as described previously [22 (link)]. After blocking with 10% normal goat serum (Sigma-Aldrich, USA), slices were incubated with primary antibodies as following: rabbit anti-GFAP (1:300; Cell Signaling Technology, USA), rabbit anti-IBA1 (1:400; Abcam, Cambridge, MA, USA), rabbit anti-beta Amyloid (1:50; Abcam, Cambridge, MA, USA), and mouse anti-Olig1 (1:100; Santa Cruz, USA). After incubated overnight at 4 °C and washed three times in 0.01 M PBS (3 × 5 min), slices were incubated with the following secondary antibodies for 1 h at room temperature: Alexa Fluor^® 594 conjugated goat anti-rabbit IgG or Alexa Fluor^® 488 conjugated goat anti-rabbit IgG or Alexa Fluor^® 488 conjugated goat anti-mouse IgG(1:1000; Cell Signaling Technology). Slices were mounted in ProLong^® Gold antifade reagent (Thermo Fisher Scientific, USA) prior to imaging.