Two-photon fluorescence correlation spectroscopy as a tool for measuring molecular diffusion within human skin
| Autor: | Vladimir Kirejev, Marica B. Ericson, Mattias Goksör, Stina Guldbrand, Carl Simonsson, Maria Smedh |
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| Rok vydání: | 2013 |
| Předmět: |
Fluorophore
Administration Topical Chemistry Pharmaceutical Analytical chemistry Pharmaceutical Science Fluorescence correlation spectroscopy Human skin In Vitro Techniques Diffusion Matrix (chemical analysis) chemistry.chemical_compound Fluorescence microscope Rhodamine B Humans Fluorescent Dyes Skin Molecular diffusion Microscopy Confocal Rhodamines Chemistry General Medicine Allergens Fluorescence Spectrometry Fluorescence Microscopy Fluorescence Biotechnology |
| Zdroj: | European Journal of Pharmaceutics and Biopharmaceutics. 84:430-436 |
| ISSN: | 0939-6411 |
| Popis: | There is a need for tools enabling quantitative imaging of biological tissue for pharmaceutical applications. In this study, two-photon fluorescence microscopy (TPM) has been combined with fluorescence correlation spectroscopy (FCS), demonstrating proof-of-principle providing quantitative data of fluorophore concentration and diffusion in human skin. Measurements were performed on excised skin exposed to either rhodamine B (RB) or rhodamine B isothiocyanate (RBITC), chosen based on their similarity in fluorescence yield and molecular weight, but difference in chemical reactivity. The measurements were performed at tissue depths in the range 0 and 20 μm, and the diffusion coefficients at skin depths 5 and 10 μm were found to be significantly different (P0.05). Overall median values for the diffusion coefficients were found to be 4.0×10(-13) m(2)/s and 2.0×10(-13) m(2)/s for RB and RBITC, respectively. These values correspond to the diffusion of a hard sphere with a volume eight times larger for RBITC compared to RB. This indicates that the RBITC have bound to biomolecules in the skin, and the measured signal is obtained from the RBITC-biomolecule complexes, demonstrating the potential of the TPM-FCS method to track molecular interactions in an intricate biological matrix such as human skin. |
| Databáze: | OpenAIRE |
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