Multiphoton tomography was used to examine xenobiotic transport in vivo. We used the photochemical properties of zinc oxide and fluorescein and multiphoton tomography to study their transport in the skin and in the rat liver in vivo. Zinc oxide nanoparticles were visualised in human skin using the photoluminescence properties of zinc oxide and either a selective emission wavelength band pass filter or a filter with fluorescence lifetime imaging (FLIM). Zinc oxide nanoparticles (30 nm) did not penetrate into human skin in vitro and in vivo and this was validated by scanning electron microscopy with X-ray photoelectron spectroscopy. Fluorescein was measured in the liver using FLIM. Fluorescein is rapidly extracted from the blood into the liver cells and then transported into the bile. It is suggested that multiphoton tomography may be of particular use in defining in vivo 4D (in both space and time) pharmacokinetics.