In this article, anionic lipophilic dye merocyanine 540(MC540) and cationic surface-active ionic liquid (SAIL) 1-octyl-3-methylimidazolium chloride (C₈mimCl) are employed to construct highly ordered fibrillar and vesicular aggregates exploiting an ionic self-assembly (ISA) strategy. It is noteworthy that the concentration of the counterions has exquisite control over the morphology, in which lowering the concentration of both the building blocks in a stoichiometric ratio of 1:1 provides a vesicle to fibril transition. Here, we have reported the concentration-controlled fibril–vesicle transition utilizing the emerging fluorescence lifetime imaging microscopy (FLIM) technique. Furthermore, we have detected this morphological transformation by means of other microscopic techniques such as field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), and cryogenic-transmission electron microscopy (cryo-TEM) to gain additional support. Besides, multiwavelength FLIM (MW-FLIM) and atomic force microscopy (AFM) techniques assist us in knowing the microheterogeneity and the height profile of the vesicles, respectively. We have replaced the SAIL, C₈mimCl, by an analogous traditional surfactant, n-octyltrimethylammonium bromide (OTAB), and it provides a discernible change in morphology similar to that of C₈mimCl, whereas 1-octanol is unable to exhibit any structural aggregation and thus reveals the importance of electrostatic interaction in supramolecular aggregate formation. However, the SAILs having the same imidazolium headgroup with different chain lengths other than C₈mimCl are unable to display any structural transition and determine the importance of the correct chain length for efficient packing of the counterions to form a specific self-assembly. Therefore, this study reveals the synergistic interplay of electrostatic, hydrophobic, and π–π stacking interactions to construct the self-assembly and their concentration-dependent morphological transition.