Lipid droplets are dynamic organelles involved in various physiological processes and their detection is thus of high importance to biomedical research. Recent reports show that AIE probes for lipid droplet imaging have the superior advantages of high brightness, large Stokes shift and excellent photostability compared to commercial dyes but suffer from the problem of having a short excitation wavelength. In this work, an AIE probe, namely TPA-BI, was rationally designed and easily prepared from triphenylamine and imidazolone building blocks for the two-photon imaging of lipid droplets. TPA-BI exhibited TICT+AIE features with a large Stokes shift of up to 202 nm and a large two-photon absorption cross-section of up to 213 GM. TPA-BI was more suitable for two-photon imaging of the lipid droplets with the merits of a higher 3D resolution, lesser photobleaching, a reduced autofluorescence and deeper penetration in tissue slices than a commercial probe based on BODIPY 493/503, providing a promising imaging tool for lipid droplet tracking and analysis in biomedical research and clinical diagnosis.