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Int. J. Mol. Sci. 2015, 16(4), 6960-6977; doi:10.3390/ijms16046960

Application of Single Molecule Fluorescence Microscopy to Characterize the Penetration of a Large Amphiphilic Molecule in the Stratum Corneum of Human Skin

1
Department of Physics, Institute of Experimental Physics, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, Germany
2
Clinical Research Center for Hair and Skin Science, Department of Dermatology, Charité-Universitaetsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany
*
Authors to whom correspondence should be addressed.
Academic Editor: Herbert Schneckenburger
Received: 15 December 2014 / Revised: 12 March 2015 / Accepted: 13 March 2015 / Published: 27 March 2015
(This article belongs to the Special Issue Laser Application in Life Sciences)
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Abstract

We report here on the application of laser-based single molecule total internal reflection fluorescence microscopy (TIRFM) to study the penetration of molecules through the skin. Penetration of topically applied drug molecules is often observed to be limited by the size of the respective drug. However, the molecular mechanisms which govern the penetration of molecules through the outermost layer of the skin are still largely unknown. As a model compound we have chosen a larger amphiphilic molecule (fluorescent dye ATTO-Oxa12) with a molecular weight >700 Da that was applied to excised human skin. ATTO-Oxa12 penetrated through the stratum corneum (SC) into the viable epidermis as revealed by TIRFM of cryosections. Single particle tracking of ATTO-Oxa12 within SC sheets obtained by tape stripping allowed us to gain information on the localization as well as the lateral diffusion dynamics of these molecules. ATTO-Oxa12 appeared to be highly confined in the SC lipid region between (intercellular space) or close to the envelope of the corneocytes. Three main distinct confinement sizes of 52 ± 6, 118 ± 4, and 205 ± 5 nm were determined. We conclude that for this amphiphilic model compound several pathways through the skin exist. View Full-Text
Keywords: single molecule fluorescence microscopy; total internal reflection fluorescence microscopy; single particle tracking; skin penetration; penetration pathways; stratum corneum; tape stripping single molecule fluorescence microscopy; total internal reflection fluorescence microscopy; single particle tracking; skin penetration; penetration pathways; stratum corneum; tape stripping
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Volz, P.; Boreham, A.; Wolf, A.; Kim, T.-Y.; Balke, J.; Frombach, J.; Hadam, S.; Afraz, Z.; Rancan, F.; Blume-Peytavi, U.; Vogt, A.; Alexiev, U. Application of Single Molecule Fluorescence Microscopy to Characterize the Penetration of a Large Amphiphilic Molecule in the Stratum Corneum of Human Skin. Int. J. Mol. Sci. 2015, 16, 6960-6977.

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