We present the contrast mechanisms of scanning electron microscopy (SEM) for visualizing the interface between carbon nanofibers (CNFs) and the underlying substrate. SEM imaging with electron beam energies higher than a certain threshold provides different image contrasts depending on whether CNFs are in contact with the substrate or suspended above the substrate. CNFs with diameters ranging from 25to250nm are examined with various electron beam energies. It is found that the threshold energy corresponds to the energy required to penetrate the CNF and its dependence on CNF diameter can be understood using the theory of electron range. This knowledge will be quite useful for interface imaging of all nanostructure devices.