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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editor: Gregory W. Faris
  • Vol. 5, Iss. 12 — Sep. 30, 2010

Two-photon fluorescence correlation microscopy combined with measurements of point spread function; investigations made in human skin

Stina Guldbrand, Carl Simonsson, Mattias Goksör, Maria Smedh, and Marica B. Ericson  »View Author Affiliations

Optics Express, Vol. 18, Issue 15, pp. 15289-15302 (2010)

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Two-photon excitation fluorescence correlation spectroscopy (TPFCS) has been applied in connection to measurements of the point spread function (PSF) for quantitative analysis of sulphorhodamine B (SRB) in excised human skin. The PSF was measured using subresolution fluorescent beads embedded in the skin specimen. The PSF, measured as full width at half maximum (FWHM) was found to be 0.41 ± 0.05 µm in the lateral direction, and 1.2 ± 0.4 μm in the axial direction. The molecular diffusion of SRB inside the skin ranged between 0.5 and 15.0 × 10−8 cm2/s. The diffusion coefficient is not dependent on depths down to 40 µm. The fluorophores were found to accumulate on the upper layers of the skin. This work is the first TPFCS study in human skin. The results show that TPFCS can be used for quantitative analyses of fluorescent compounds in human skin.

© 2010 OSA

OCIS Codes
(170.7050) Medical optics and biotechnology : Turbid media
(180.2520) Microscopy : Fluorescence microscopy
(300.6410) Spectroscopy : Spectroscopy, multiphoton

ToC Category:

Original Manuscript: April 8, 2010
Revised Manuscript: May 31, 2010
Manuscript Accepted: June 6, 2010
Published: July 2, 2010

Virtual Issues
Vol. 5, Iss. 12 Virtual Journal for Biomedical Optics

Stina Guldbrand, Carl Simonsson, Mattias Goksör, Maria Smedh, and Marica B. Ericson, "Two-photon fluorescence correlation microscopy combined with measurements of point spread function; investigations made in human skin," Opt. Express 18, 15289-15302 (2010)

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