Zinc oxide nanoparticles (ZnO NPs) are commonly used in sunscreens to reduce the risk of skin cancer by blocking ultraviolet radiation. ZnO NPs absorption through the transdermal route may not cause high health risk as inhalation or ingestion. However, in practical usage of sunscreens and cosmetics, ZnO NPs are topically applied to a large area of skin with long periods hence the potential absorption amount of ZnO NPs is still need to be concerned. Therefore, if the ZnO NPs are able the pass the barrier of normal skin, the pathways of transdermal delivery and the factors of enhancements become important issues. In this work, multiphoton microscopy provides us a non-invasive visualization of ZnO NPs in skin. Moreover, we quantitatively analyzed the enhancement of oleic acid and ethanol. Due to the fact that photoluminance of ZnO NPs spectrally overlaps autofluorence from skin stratum corneum (SC) and high turbidity of both ZnO NPs and SC, it is difficult to resolve the distribution of ZnO NPs in skin by using fluorescence microscopy. In this work, the second harmonic generation (SHG) signals from ZnO NPs which double the frequency of excitation source to characterize the delivery pathways and penetration depth in skin. Moreover, we quantitatively compare the ZnO NPs delivery efficiency in normal skin and in skins with three chemically enhancing conditions: ethanol, oleic acid and the combination of ethanol and oleic acid.