Two-photon fluorescence correlation spectroscopy as a tool for measuring molecular diffusion within human skin

Guldbrand, Stina,1970Gothenburg University,Göteborgs universitet,Institutionen för fysik (GU),Department of Physics (GU) (författare)

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 pm, and the diffusion coefficients at skin depths 5 and 10 mu m were found to be significantly different (P < 0.05). Overall median values for the diffusion coefficients were found to be 4.0 x 10(-13) m(2)/s and 2.0 x 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.

MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper Farmaceutiska vetenskaper hsv//swe
MEDICAL AND HEALTH SCIENCES Basic Medicine Pharmaceutical Sciences hsv//eng
Two-photon fluorescence microscopy
Fluorescence correlation spectroscopy
Diffusion coefficient
living cells
microscopy
visualization
transport

Kirejev, Vladimir,1984Gothenburg University,Göteborgs universitet,Institutionen för fysik (GU),Department of Physics (GU)(Swepub:gu)xkirvl (författare)
Simonsson, Carl,1976Gothenburg University,Göteborgs universitet,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology(Swepub:gu)xsimca (författare)
Goksör, Mattias,1975Gothenburg University,Göteborgs universitet,Institutionen för fysik (GU),Department of Physics (GU)(Swepub:gu)xgokma (författare)
Smedh, Maria,1968Gothenburg University,Göteborgs universitet,Core Facilities, Centre for Cellular Imaging,Core Facilities, Centre for Cellular Imaging(Swepub:gu)xsmema (författare)
Ericson, Marica B,1974Gothenburg University,Göteborgs universitet,Institutionen för fysik (GU),Department of Physics (GU)(Swepub:gu)xemarq (författare)
Göteborgs universitetInstitutionen för fysik (GU) (creator_code:org_t)

Ingår i:European Journal of Pharmaceutics and Biopharmaceutics: Elsevier BV84:2, s. 430-4360939-6411

Av författaren/redakt...: Guldbrand, Stina ...; Kirejev, Vladimi ...; Simonsson, Carl, ...; Goksör, Mattias, ...; Smedh, Maria, 19 ...; Ericson, Marica ...

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MEDICIN OCH HÄLSOVETENSKAP: MEDICIN OCH HÄLS ...; och Medicinska och f ...; och Farmaceutiska ve ...

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