Ckx41 culture microscope

Melan-a melanocytes were seeded in 24-well plates and maintained for 24 h. The cells were then rinsed in DPBS and treated with samples in RPMI-1640 containing 2% FBS for 3 days. The cells were observed in bright field using an Olympus CKX41 culture microscope (Olympus, Japan), and images were photographed using a DMC camera (INS Industry, Korea) and DMC advanced software adapted to the microscope. Evaluation of melanosome aggregation was performed by counting cells with perinuclear melanosome aggregates in three random microscopic fields per well at x200 magnification. Values are presented as the mean ± SD from three wells (n = 3).

After electrophysiological recording, a 2% fluorogold suspension was prepared by dissolving fluorogold in distilled water, stored at 4 °C in the dark, and directly injected using a Hamilton micro-syringe into the common peroneal nerve and posterior tibial nerve. Five days later, the rats were trans-cranially perfused sequentially with 200 mL of 0.9% saline, followed by cold 4% paraformaldehyde in 0.1 M PBS. Next, L4 and L5 DRGs on the same side of the injury were dissected and soaked in 4% paraformaldehyde for post-fixation overnight, and in 30% phosphate-buffered sucrose solution for additional overnight. DRGs of 40-μm thickness were then obtained from longitudinal sections of the spinal cord. After drying, the section was mounted and observed under an ultraviolet fluorescence microscope (Olympus Ckx41 Culture Microscope).