AbstractDeveloping a convenient method to determine the complete structure of single‐walled carbon nanotubes (SWNTs) is important to achieve the fully controlled growth of this nanomaterial. However, approaches that can identify handedness at the atomic level with simple equipment, operation, and data analysis are still lacking. Here, the SWNTs/graphene (Gr) vertical heterostructures are artificially constructed with aligned interfaces to realize the lattice interpretation of SWNT upper and lower walls separately by only one transmission electron microscopy image, thus transforming the 3D handedness information to projected 2D space. Gr displays prominent out‐of‐plane deformation at the interface, promoting the energetic advantage for the aligned interface construction. The interfacial alignment between the SWNT and Gr shows no obvious dependence on either the helical angle or diameter of SWNTs. The half‐wrapping of SWNTs by deformed Gr also triggers diversified alterations in electronic structures based on theoretical calculations. 27 specimens with SWNTs prepared by two disparate methods are examined, implying equal handedness distribution in the randomly aligned SWNTs grown on quartz and potential handedness enrichment in horizontal SWNT arrays grown on a‐sapphire. This work provides a simple strategy for chiral discrimination and lays a characterization foundation for handedness‐selective growth of nanomaterials.