Aggregation-induced emission (AIE)-active fluorescent nanoprobes for lipid droplets (LDs) have great potential for monitoring the physiological dynamic evolution of LDs due to their high biocompatibility and excellent photostability. However, hydrophobic LDs probe exhibit weak intermolecular interactions in aqueous media, which usually leads to disordered spontaneous aggregation and uncontrollable microstructure, thereby restricting the controllable fabrication of high-performance LDs fluorescent organic nanoprobes (FONs). To address this issue, an AIE-active probe CPB-BTH-Hex with solvent polarity sensitivity was rationally constructed via a molecular engineering strategy. By improving molecular planarity, the intermolecular π···π interactions were effectively strengthened, enabling the formation of highly ordered pre-aggregates before encapsulation and achieving precise regulation of aggregate morphology and structure. Single-crystal structure analysis reveals that the probe adopts J-aggregation in the crystalline state, but accompanied by blue-shifted in absorption and emission spectra. Theoretical calculations combined with detailed spectroscopic investigations reveal that the probe form aggregates with magic-angle stacking in DMSO/H₂O mixtures containing a moderate water fraction. Meanwhile, the twist intramolecular charge transfer (TICT) featuring by overall molecular distortion and local parallel alignment arises from variations in molecular torsional angles, combined with magic-angle stacking collectively account for the AIE behavior of the probe in aqueous media, endowing it with a unique fluorescence enhancement effect. After encapsulation with poly(lactic-co-glycolic acid) (PLGA), the resulting nanoprobe exhibits excellent bioimaging performance. It realizes long-term and stable dynamic tracking of intracellular LDs and clearly visualizes the fusion and fission processes of LDs in living cells, providing a reliable visual tool for exploring the biological functions of lipid droplets.