The biomimetic approach on the cuticular spinning nozzles of the major ampullate silk glands in the golden-web spider Nephila calvata has been attempted using various visualizing techniques of light and electron microscopes to improve the design of spinning nozzle for producing synthetic nanofibers spun from electrospinning apparatus. The major ampullate spigot which has the most effective nozzle system to produce nanofibers for dragline silk with high strength and elasticity is connected via the bullet type spigot on anterior spinneret with flexible terminal segment. The excretory duct which transports the liquid silk feedstock from ampulla to spigot is divided into 3 limbs by loops back on itself to form an S-shape morphology that is bundled in connective tissue. Final diameter of the nanofibers at nozzle was dramatically reduced by gradual narrowing of duct cuticle less than 10 times comparing to its original size of funnel region. Moreover, the funnel has a characteristic cuticular organization with porous microstructure which seems to be related to water removal from feedstock of silk precursors. High magnification electron micrographs also reveal the presence of the spiral grooves on the surface of the cuticular intima near the valve which presumed to reduce friction during rapid flow of liquid silk.