Mechanically responsive materials that are able to sense and respond to external stimuli have important applications in soft robotics and the formation of artificial muscles, such as intelligent electronics, prosthetic limbs, comfort-adjusting textiles and miniature actuators for microfluidics. However, previous artificial muscles based on polymer materials are insufficient in generating large actuations, fast responses, diverse deformation modes and high cycle performances. To this end, carbon nanotubes (CNTs) are proposed as promising candidates to be assembled into artificial muscles, as they are lightweight, robust and have high surface-to-volume ratios. This protocol describes a reproducible biomimetic method for preparing a family of hierarchically arranged helical fiber (HHF) actuators that are responsive to solvents and vapors. These HHFs are produced through helical assembly of CNTs into primary fibers and further twisting of the multi-ply primary fibers into a helical structure. A large number of nanoscale gaps between the CNTs and micron-scale gaps between the primary fibers ensure large volume changes and fast responses upon the infiltration of solvents and vapors (e.g., water, ethanol, acetone and dichloromethane) by capillarity. The modes of shape transformations can be modulated precisely by controlling how the CNTs are assembled into primary fibers, multi-ply primary fibers, HHFs and hierarchical springs. This protocol provides a prototype for preparing actuators with different fiber components. The overall time required for the preparation of HHF actuators is 17 h.