Quantifying Blood-Brain Barrier Leakage

Blood–brain barrier leakage was assessed via EB staining at 24 h after ICH. Two percent EB solution (8 mL/kg, Sigma–Aldrich) was applied through femoral vein after anesthetization. Two hours later, the rats received transcardial perfusion with 0.1M PBS. Next, injured brain hemisphere was collected and homogenized in N, N-dimethylformamide. The sample was incubated in water bath (50°C) for 48 h and centrifuged at 12,000 × g for 30 min. Finally, the supernatant was collected and measured at 620 nm with a spectrophotometer (2,000°C, Thermo Fisher) (Zhao et al., 2016 (link)).