We investigated the effect of PEGylated liposomes (PLs) containing a terpene on the penetration of a hydrophilic compound through porcine skin. PLs composed of N -(carbonyl-methoxypolyethyleneglycol-2000)–1,2-distearoyl- sn -glycero-3-phosphoethanolamine (PEG₂₀₀₀–DSPE), the sodium salt of PEG₂₀₀₀–DSPE, phosphatidylcholine (PC), cholesterol (Chol), Tween 20, and d -limonene were prepared as carriers for fluorescein sodium (NaFI). The physicochemical characteristics of PLs and their effects on in vitro skin penetration were evaluated. Tape stripping was used to evaluate NaFI deposition in skin layers, and confocal laser scanning microscopy (CLSM) was used to investigate the depth of skin penetration and the pathways used by NaFI-loaded vesicles. PLs containing d -limonene were smaller and conferred higher entrapment efficiency and skin penetration on NaFI than did PLs and conventional liposomes (CLs). The deposition of NaFI from PLs with d -limonene was greater in epidermis and dermis (6.10±1.74 µg) than stratum corneum (2.06±0.47 µg). CLSM images revealed that NaFI penetrated into the deepest skin layer with maximum fluorescence intensity. NaFI penetrated deeper (180 µm) in follicular than nonfollicular regions (145 µm), suggesting a transfollicular pathway predominates in skin penetration by NaFI-loaded PLs. In conclusion, grafting PEG onto ultra-deformable liposomes may enhance transdermal NaFI delivery and may be used as a carrier to prolong liposome circulation time.