title: Application of single molecule fluorescence microscopy to characterize the penetration of a large amphiphilic molecule in the stratum corneum of human skin.
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.
authors.affiliation: Department of Physics, Institute of Experimental Physics, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, Germany. pierre.volz@fu-berlin.de.
authors.forename: Pierre
authors.initials: P
authors.lastname: Volz
authors.affiliation: Department of Physics, Institute of Experimental Physics, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, Germany. alexander.boreham@fu-berlin.de.
authors.forename: Alexander
authors.initials: A
authors.lastname: Boreham
authors.affiliation: Department of Physics, Institute of Experimental Physics, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, Germany. alexander.wolf@fu-berlin.de.
authors.forename: Alexander
authors.initials: A
authors.lastname: Wolf
authors.affiliation: Department of Physics, Institute of Experimental Physics, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, Germany. tai-yang.kim@fu-berlin.de.
authors.forename: Tai-Yang
authors.initials: TY
authors.lastname: Kim
authors.affiliation: Department of Physics, Institute of Experimental Physics, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, Germany. jens.balke@fu-berlin.de.
authors.forename: Jens
authors.initials: J
authors.lastname: Balke
authors.affiliation: Clinical Research Center for Hair and Skin Science, Department of Dermatology, Charité-Universitaetsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany. janna.frombach@charite.de.
authors.forename: Janna
authors.initials: J
authors.lastname: Frombach
authors.affiliation: Clinical Research Center for Hair and Skin Science, Department of Dermatology, Charité-Universitaetsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany. sabrina.hadam@charite.de.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Hadam
authors.affiliation: Clinical Research Center for Hair and Skin Science, Department of Dermatology, Charité-Universitaetsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany. zahra.afraz@charite.de.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Afraz
authors.affiliation: Clinical Research Center for Hair and Skin Science, Department of Dermatology, Charité-Universitaetsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany. fiorenza.rancan@charite.de.
authors.forename: Fiorenza
authors.initials: F
authors.lastname: Rancan
authors.affiliation: Clinical Research Center for Hair and Skin Science, Department of Dermatology, Charité-Universitaetsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany. ulrike.blume-peytavi@charite.de.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Blume-Peytavi
authors.affiliation: Clinical Research Center for Hair and Skin Science, Department of Dermatology, Charité-Universitaetsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany. annika.vogt@charite.de.
authors.forename: Annika
authors.initials: A
authors.lastname: Vogt
authors.affiliation: Department of Physics, Institute of Experimental Physics, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, Germany. ulrike.alexiev@fu-berlin.de.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Alexiev
mesh_terms: D004817:Epidermis / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000493:pharmacokinetics*; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D012869:Skin Absorption
chemical_list: D005456:Fluorescent Dyes
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