Vt1000 s vibrating blade vibratome

Golgi-cox staining was performed as we described previously [10 (link)]. Briefly, brains were perfused with 20 ml PBS by transcardiac perfusion under isoflurane anesthesia, after which brains were removed and processed using the FD Rapid GolgiStain™ Kit (FD NeuroTechnologies, Columbia, MD) following the manufacturer’s instructions. A Leica VT1000 S Vibrating-blade vibratome was used to cut 130 to 150μm coronal sections of the fixed brains. The sections were cut in artificial cerebrospinal fluid (aCSF) comprised of 240 mM sucrose, 3.3 mM KCl, 26 mM NaHCO₃, 1.3 mM MgSO₄·7H₂O, 1.23 mM NaH₂PO₄, 11 mM D-glucose, and 1.8 mM CaCl₂. Sections were mounted on gelatin-coated slides and dried in the dark in a fume hood for at least 24 h prior to staining.

Under isoflurane anesthesia, brains were perfused with 20 ml PBS by transcardiac perfusion. Brains were removed and processed using the FD Rapid GolgiStain™ Kit (FD NeuroTechnologies, Columbia MD) following the manufacturer’s instructions. A Leica VT1000 S Vibrating-blade vibratome was used to cut coronal sections of the fixed brains. Section thickness was 130 to 150 μm, the speed adjustment was 0.175 mm/s (scale setting 4 to 6), and the vibration frequency was 30 Hz (scale setting 3). The sections were cut in artificial cerebrospinal fluid (240 mM sucrose, 3.3 mM KCl, 26 mM NaHCO₃, 1.3 mM MgSO₄·7H₂O, 1.23 mM NaH₂PO₄, 11 mM D-glucose, and 1.8 mM CaCl₂.) and then mounted on gelatin-coated slides with a drop of ‘C’ solution from the FD Rapid GolgiStain™ Kit. After mounting sections on the slide, the remaining ‘C’ solution was wiped away with a strip of filter paper to ensure that the slides totally dried and that the sections remained attached during the staining process. The slides were dried in the dark in a fume hood for at least 24 h prior to staining.