Objective:To establish an effective model and evaluation system for lysolecithin (LPC) induced demyelination in dorsal funiculus of cervical spinal segment in the adult mice. Methods:The adult mice were injected with 0.4 μl 1% LPC or normal saline into the dorsal funiculus of the 5th cervical (C5) spinal segment using glass-needle-capped syringe under the microscope. One week post surgery,mice were sacrificed and perfused,and segments of C4 to C6 were dissected and embedded in paraffin. Serial coronal sections were stained with toludine blue. The 3D cartoon of demyelinated area of total serial sections was rebuilt,and the volume of the demyelinated area was measured by Amira 4.1.1 software. The demyelinated axons were also observed by electron microscope. Results:No obvious motor and sensory abnormity were found after mice recovered from the anesthesia. Toludine blue staining result showed that there was the light blue or pale demyelination area around the injection point,and the volume of which was about (1.41 ± 0.05) × 106 μm3. The demyelinated axons were confirmed by electron microscopy observation. Conclusion:Precise injection of LPC into the C5 dorsal funiculus of mice can stably induce the axonal demyelination.