Abstract We investigated the microstructures of carbon nanotube (CNT) films and the effect of CNT length on their mechanical performance. 230 μm-, 300 μm-, and 360 μm- long CNTs were grown and used to fabricate CNT films by a winding process. Opposite from the length effect on CNT fibers, it has been found that the mechanical properties of the CNT films decrease with increasing CNT length. Without fiber twisting, short CNTs tend to bundle together tightly by themselves in the film structure, resulting in an enhanced packing density; meanwhile, they also provide a high degree of CNT alignment, which prominently contributes to high mechanical properties of the CNT films. When CNTs are long, they tend to be bent and entangled, which significantly reduce their packing density, impairing the film mechanical behaviors severely. It has also been unveiled that the determinant effect of the CNT alignment on the film mechanical properties is more significant than that of the film packing density. These findings provide guidance on the optimal CNT length when attempting to fabricate high-performance macroscopic CNT assemblies.