Histological Analysis of Ischemic Brains

Mice were euthanized within 24 h following MCAO or 4–5 weeks post sham surgery for preparation of brain histological sections (El-Hayek et al., 2011 (link); Wang et al., 2015 (link); Wu et al., 2015 (link)). Briefly, mice were anesthetized via an intra-peritoneal injection of sodium pentobarbital (100 mg/kg) and trans cardially infused with 10% neutral buffered formalin solution. Brains were removed and further fixed in a hypertonic (with 20% sucrose) formalin solution. Cryostat coronal sections of 30 μm thickness were obtained and stained with cresyl violet. Images of brain sections were obtained using a slide scanner (Aperio digital pathology slide scanner AT2, Leica) at 20× magnification and analyzed using ImageScope (Leica) and ImageJ (National Institute of Health, Bethesda, MD, USA) software.