Quantifying Glial Cells Post-Hypoxic Injury

At 72 h post-HI, the brains were collected, sectioned into 4 µm slices in the region containing the infarct lesion. All brain slices were obtained from independent experiments and assays were performed from brain sections of similar anatomical locations. Following our previous procedures,7 (link) These slices were stained with adaptor molecule-1 (Iba-1) (ab178847, Abcam, Cambridge, UK) or glial fibrillary acidic protein (GFAP) (60190-1-Ig, Proteintech Group, Rosemont, IL, USA). The slices were incubated with a fluorescently labeled secondary antibody for 30 min at 37°C in the dark followed by DAPI staining. Fluorescent images were observed using fluorescent microscopy and Image-Pro Plus 6.0 software was used to analyze the number of Iba-1⁺ and GFAP ⁺ cells in the right cortex.