Cresyl violet staining

Frozen sections of mouse brain were dried at room temperature for 5 minutes and washed with 1XPBS for 5 times, then incubated in graded ethanol (95% ethanol 15 minutes, 70% ethanol 1 minute, 50% ethanol 1 minute). After washing with distilled water, the brain tissues were stained with 0.5% cresyl violet acetate (Sigma-Aldrich) solution for 12 minutes and passed through distilled water (1 minute), 50% ethanol (1 minute), 70% ethanol (2 minutes), 95% ethanol (2 times 2 minutes), 100% ethanol (1 minute) and finally cleaned with xylene (5 minutes). Stained slides were mounted with DPX mounting medium for histological analysis (Sigma-Aldrich).