Label free microscopy provides insights into nanocarrier and drug penetration characteristics in human and murine skin. Selective probing of drugs and nanocarriers in skin, using X-ray spectromicroscopy, was conducted to determine the drug and nanocarrier distribution, penetration pathways, and concentration in skin. A unique stimulated Raman microscopy setup is introduced. Results on the protein, lipid, and water distribution in human skin together with results from life science microscopy are reported.