2.4. Intraepidermal Nerve Fiber (IENF) Density Determination

According to reports, the lack of IENFs plays a key role in neuropathy caused by various chemotherapy drugs (such as oxaliplatin) [14]. On the last day of the sixth week, the plantar skin tissues of the mice were removed, fixed in a periodate-lysine-paraformaldehyde (PLP) solution for 18-22 hours, and then transferred to a 30% sucrose-containing PBS solution to dehydrate until the tissue subsided, and frozen sections were made within one week. The tissue was embedded with optimal cutting temperature compound (OCT) and sectioned frozen (continuously cut perpendicular to the epidermis) with a thickness of approximately 30 μm. The sections were subsequently stained, and the state of the epidermal nerve fibers was observed under an inverted microscope.