Differentiation of peripheral nerve functions and properties with spectral analysis and Karnovsky-Roots staining: A preliminary study

Objective: The purpose of this study was to explore the possibility for analyzing and differentiating between motor and sensory functions of peripheral nerve axons using spectral technology. Methods: 10 μm slide section of S1 anterior and posterior rabbit spinal nerve roots were made and then stained with Karnovsky-Roots method for molecular hyperspectral imaging microscopy analysis. In addition, Raman spectra data of nerve axons on each slide was collected after Karnovsky-Roots staining for 30 minutes. Results: Motor axons were differentiated from sensory axons in a nerve axon section hyperspectral image via Spectral angle mapper algorithm. Raman scatterings could be detected near 2110 cm⁻¹, and 2155 cm⁻¹ in motor axons after Karnvosky-Roots staining. The value of I₂₁₀₀/I₁₄₄₀ in motor axons are significantly different (P0.001) than in sensory axons after staining for 30 minutes. Conclusions: Motor and sensory nerve axons can be differentiated from their counterparts in 30 minutes by using Raman micro-spectroscopy analysis assisted with Karnovsky-Roots staining.