OSA's Digital Library

Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 18, Iss. 8 — Apr. 12, 2010
  • pp: 7857–7871

Multiphoton excitation characteristics of cellular fluorophores of human skin in vivo

Hans Georg Breunig, Hauke Studier, and Karsten König  »View Author Affiliations

Optics Express, Vol. 18, Issue 8, pp. 7857-7871 (2010)

View Full Text Article

Enhanced HTML    Acrobat PDF (3928 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



In vivo multiphoton tomography with a wavelength-tunable femtosecond laser has been performed to investigate the autofluorescence intensity of major endogenous fluorophores of human skin in dependence on the excitation wavelength. In high-resolution multiphoton images of different skin layers, clear trends were found for fluorophores like keratin, NAD(P)H, melanin as well as for the elastin and collagen networks. The analysis of the measurements is supplemented by additional measurements of fluorescence lifetime imaging and signal-decay curves by time-correlated single-photon counting.

© 2010 OSA

OCIS Codes
(170.1610) Medical optics and biotechnology : Clinical applications
(170.4580) Medical optics and biotechnology : Optical diagnostics for medicine
(170.5810) Medical optics and biotechnology : Scanning microscopy
(190.1900) Nonlinear optics : Diagnostic applications of nonlinear optics
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: December 17, 2009
Revised Manuscript: February 8, 2010
Manuscript Accepted: February 9, 2010
Published: March 31, 2010

Virtual Issues
Vol. 5, Iss. 8 Virtual Journal for Biomedical Optics

Hans Georg Breunig, Hauke Studier, and Karsten König, "Multiphoton excitation characteristics of cellular fluorophores of human skin in vivo," Opt. Express 18, 7857-7871 (2010)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248(4951), 73–76 (1990). [CrossRef] [PubMed]
  2. B. R. Masters, P. T. So, and E. Gratton, “Multiphoton excitation fluorescence microscopy and spectroscopy of in vivo human skin,” Biophys. J. 72(6), 2405–2412 (1997). [CrossRef] [PubMed]
  3. P. T. So, C. Y. Dong, B. R. Masters, and K. M. Berland, “Two-photon excitation fluorescence microscopy,” Annu. Rev. Biomed. Eng. 2(1), 399–429 (2000). [CrossRef]
  4. K. König, “Clinical multiphoton tomography,” J Biophotonics 1(1), 13–23 (2008). [CrossRef]
  5. K. M. Hanson and C. J. Bardeen, “Application of nonlinear optical microscopy for imaging skin,” Photochem. Photobiol. 85(1), 33–44 (2009). [CrossRef] [PubMed]
  6. K. König and I. Riemann, “High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution,” J. Biomed. Opt. 8(3), 432–439 (2003). [CrossRef] [PubMed]
  7. E. Dimitrow, I. Riemann, A. Ehlers, M. J. Koehler, J. Norgauer, P. Elsner, K. König, and M. Kaatz, “Spectral fluorescence lifetime detection and selective melanin imaging by multiphoton laser tomography for melanoma diagnosis,” Exp. Dermatol. 18(6), 509–515 (2009). [CrossRef] [PubMed]
  8. E. Dimitrow, M. Ziemer, M. J. Koehler, J. Norgauer, K. König, P. Elsner, and M. Kaatz, “Sensitivity and specificity of multiphoton laser tomography for in vivo and ex vivo diagnosis of malignant melanoma,” J. Invest. Dermatol. 129(7), 1752–1758 (2009). [CrossRef] [PubMed]
  9. M. J. Koehler, K. König, P. Elsner, R. Bückle, and M. Kaatz, “In vivo assessment of human skin aging by multiphoton laser scanning tomography,” Opt. Lett. 31(19), 2879–2881 (2006). [CrossRef] [PubMed]
  10. M. J. Koehler, A. Preller, N. Kindler, P. Elsner, K. König, R. Bückle, and M. Kaatz, “Intrinsic, solar and sunbed-induced skin aging measured in vivo by multiphoton laser tomography and biophysical methods,” Skin Res. Technol. 15(3), 357–363 (2009). [CrossRef] [PubMed]
  11. K. Schenke-Layland, I. Riemann, O. Damour, U. A. Stock, and K. König, “Two-photon microscopes and in vivo multiphoton tomographs--powerful diagnostic tools for tissue engineering and drug delivery,” Adv. Drug Deliv. Rev. 58(7), 878–896 (2006). [CrossRef] [PubMed]
  12. F. Stracke, B. Weiss, C. M. Lehr, K. König, U. F. Schaefer, and M. Schneider, “Multiphoton microscopy for the investigation of dermal penetration of nanoparticle-borne drugs,” J. Invest. Dermatol. 126(10), 2224–2233 (2006). [CrossRef] [PubMed]
  13. K. König, A. Ehlers, F. Stracke, and I. Riemann, “In vivo drug screening in human skin using femtosecond laser multiphoton tomography,” Skin Pharmacol. Physiol. 19(2), 78–88 (2006). [CrossRef] [PubMed]
  14. J. Chen, S. Zhuo, T. Luo, X. Jiang, and J. Zhao, “Spectral characteristics of autofluorescence and second harmonic generation from ex vivo human skin induced by femtosecond laser and visible lasers,” Scanning 28(6), 319–326 (2006). [CrossRef] [PubMed]
  15. J. A. Palero, H. S. de Bruijn, A. van der Ploeg van den Heuvel, H. J. Sterenborg, and H. C. Gerritsen, “Spectrally resolved multiphoton imaging of in vivo and excised mouse skin tissues,” Biophys. J. 93(3), 992–1007 (2007). [CrossRef] [PubMed]
  16. C. Xu, W. Zipfel, J. B. Shear, R. M. Williams, and W. W. Webb, “Multiphoton fluorescence excitation: new spectral windows for biological nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 93(20), 10763–10768 (1996). [CrossRef] [PubMed]
  17. L. H. Laiho, S. Pelet, T. M. Hancewicz, P. D. Kaplan, and P. T. So, “Two-photon 3-D mapping of ex vivo human skin endogenous fluorescence species based on fluorescence emission spectra,” J. Biomed. Opt. 10(2), 024016 (2005). [CrossRef] [PubMed]
  18. S. Huang, A. A. Heikal, and W. W. Webb, “Two-photon fluorescence spectroscopy and microscopy of NAD(P)H and flavoprotein,” Biophys. J. 82(5), 2811–2825 (2002). [CrossRef] [PubMed]
  19. W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100(12), 7075–7080 (2003). [CrossRef] [PubMed]
  20. P. T. C. S. Barry R. Masters, ed., Handbook of Biomedical Nonlinear Optical Microscopy (Oxford University Press, Inc., 2008).
  21. R. M. Williams, W. R. Zipfel, and W. W. Webb, “Interpreting second-harmonic generation images of collagen I fibrils,” Biophys. J. 88(2), 1377–1386 (2005). [CrossRef]
  22. M. S. Roberts, M. J. Roberts, T. A. Robertson, W. Sanchez, C. Thorling, Y. H. Zou, X. Zhao, W. Becker, and A. V. Zvyagin, “In vitro and in vivo imaging of xenobiotic transport in human skin and in the rat liver,” J. Biophoton. 1(6), 478–493 (2008). [CrossRef]
  23. S. E. Cross, B. Innes, M. S. Roberts, T. Tsuzuki, T. A. Robertson, and P. McCormick, “Human skin penetration of sunscreen nanoparticles: in-vitro assessment of a novel micronized zinc oxide formulation,” Skin Pharmacol. Physiol. 20(3), 148–154 (2007). [CrossRef] [PubMed]
  24. W. Becker, A. Bergmann, M. A. Hink, K. König, K. Benndorf, and C. Biskup, “Fluorescence lifetime imaging by time-correlated single-photon counting,” Microsc. Res. Tech. 63(1), 58–66 (2004). [CrossRef]
  25. A. Pena, M. Strupler, T. Boulesteix, and M. Schanne-Klein, “Spectroscopic analysis of keratin endogenous signal for skin multiphoton microscopy,” Opt. Express 13(16), 6268–6274 (2005). [CrossRef] [PubMed]
  26. A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. U.S.A. 99(17), 11014–11019 (2002). [CrossRef] [PubMed]
  27. K. Teuchner, W. Freyer, D. Leupold, A. Volkmer, D. J. Birch, P. Altmeyer, M. Stücker, and K. Hoffmann, “Femtosecond two-photon excited fluorescence of melanin,” Photochem. Photobiol. 70(2), 146–151 (1999). [PubMed]
  28. A. Zoumi, X. Lu, G. S. Kassab, and B. J. Tromberg, “Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy,” Biophys. J. 87(4), 2778–2786 (2004). [CrossRef] [PubMed]
  29. A. Ehlers, I. Riemann, M. Stark, and K. König, “Multiphoton fluorescence lifetime imaging of human hair,” Microsc. Res. Tech. 70(2), 154–161 (2007). [CrossRef]

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited