A biodegradable, multilayer nanofiber structure has been successfully developed by using a self-designed and fabricated triaxial electrospinning system using gelatin as the sheath and core layers and poly(epsilon-caprolactone) (PCL) as the middle layer. The triaxial structure was investigated by confocal fluorescence microscopy (CFM) and focused ion beam and field-emission scanning electron microscopy (FIB-FESEM). The ability to fabricate the multilayered nanofibers efficiently with different biodegradable polymers will enable this triaxial electrospinning technique to have wider applications in biotechnology.