Rapid Golgi Staining of Brain Hemispheres

As previously reported, brain hemispheres, obtained as described in the animals section, were stained using the FD Rapid Golgi Stain Kit (PK401, FD Neurotechnologies, Columbia, MD, USA) and according to the manufacturer’s instructions [42 (link)]. Tissue slices were impregnated in chromate mixture of Solution A (potassium dichromate and mercuric chloride) and Solution B (potassium chromate). After 24 h, chromate solution was replaced gently without disturbing the tissue, and then left in dark for 15 days. Subsequently, tissue slices were immersed in Solution C for 24 h. After 24 h, Solution C was replaced, according to the manufacturer’s instructions. These brains were sliced in 30 μm sections, mounted three per slide on gelatin-coated slides, sequentially for each animal by FD Neurotechnologies. For further microscopic assessments, the slides were shipped back to our lab after being processed by FD Neurotechnologies. Slides were stored in darkness.