Cerebral Vascular Morphology Analysis

The rat brains collected at different time points after MCAO were harvested and sectioned at 10 µm in thickness. Following fixation with 4% paraformaldehyde, the brain slice was incubated in 100 μL blocking serum (10% fetal bovine serum in 0.2% PBST) for 1 h. Cerebral vasculature was stained with primary antibody anti-vWF (AB7356, 1:200, Milipore, Burlington, MA, USA), followed by Alexa Fluor 594 conjugated secondary antibody. Images were visualized by a confocal microscope (Fluoview BX61, Olympus, Tokyo, Japan). ImageJ was used to capture the change in vascular morphology. The total vascular area per image was quantified by ImageJ, and the average vascular diameter was determined using the shortest Feret diameter (Feret Min) as described previously [42 (link)].