A family of asymmetric cholesterol-based fluorescent organogelators containing naphthalimide, connected by two acylamines and different alkyl-chain spacers, have been designed and prepared. These compounds can gelate a variety of organic solvents with both ultrasound stimuli and general sol-gel processes. The self-assembly and gelling properties of the compounds depend on the length of the alkyl chains and can be controlled by ultrasound stimuli and renewed by a thermodynamic process. The morphologies and surface wetabilities of the xerogels prepared from these gelators are strongly affected by environmental stimuli. The mechanism of the process was investigated by confocal laser scanning microscopy, transmission or scanning electron microscopy, wide-angle X-ray scattering analysis, and rheological experiments. The studies reveal that the cooperation and relative competition of multiple intermolecular interactions, influenced by the sonication or thermal stimulus, are the main contributors for the aggregation or nucleation processes; this results in the macrodifferences in morphology and surface properties. These results provide a deeper understanding of the intermediate transition state of the gel during use of an ultrasound stimulus.