The lack of tools particularly designed for the quantification of the fiber morphology in nonwovens, especially the multi-level structured fibers, is the main reason for the limited research studies on the establishment of realistic nonwoven structure. In this study, two polymers, cellulose acetate (CA) and thermoplastic polyurethane (TPU), which have different molecular flexibility, were chosen to produce nonwovens with helical nanofibers. Focusing on the nonwovens with helical fibers, a soft package was developed to characterize fiber morphologies, including fiber orientation, helix diameter, and curvature of helix. The novelty of this study is the proposal of a method for the characterization of nanofibrous nonwovens with special fiber shape (helical fibers) which can be used for curve fibers. The characterization results for the helical-fiber nonwoven sample and the nonwoven sample with straight fibers were compared and analyzed.