OSA's Digital Library

Applied Optics

Applied Optics


  • Vol. 42, Iss. 16 — Jun. 1, 2003
  • pp: 3321–3329

Penetration depth of single-, two-, and three-photon fluorescence microscopic imaging through human cortex structures: Monte Carlo simulation

Xiaoyuan Deng and Min Gu  »View Author Affiliations

Applied Optics, Vol. 42, Issue 16, pp. 3321-3329 (2003)

View Full Text Article

Enhanced HTML    Acrobat PDF (212 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



Penetration depth is investigated in terms of the performance of transverse image resolution and signal level in human cortex under single-, two-, and three-photon fluorescence microscopy. Simulation results show that, in a double-layer human cortex structure consisting of gray and white matter media, the signal level is strongly affected by the existence of the white matter medium under three-photon excitation. Compared with three-photon excitation, two-photon excitation keeps a better signal level and sacrifices a slight degradation in image resolution. In a thick gray matter medium, a penetration depth of 1500 μm with a near-diffraction-limited resolution is obtainable under three-photon excitation. It is also demonstrated that the numerical aperture has a slight influence on image resolution and signal level under two- and three-photon excitation because of the nonlinear nature in the excitation process.

© 2003 Optical Society of America

OCIS Codes
(170.3660) Medical optics and biotechnology : Light propagation in tissues
(180.2520) Microscopy : Fluorescence microscopy
(290.4020) Scattering : Mie theory

Original Manuscript: November 13, 2002
Revised Manuscript: January 21, 2003
Published: June 1, 2003

Xiaoyuan Deng and Min Gu, "Penetration depth of single-, two-, and three-photon fluorescence microscopic imaging through human cortex structures: Monte Carlo simulation," Appl. Opt. 42, 3321-3329 (2003)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. P. S. Andersson, S. Montan, S. Svanberg, “Multispectral system for medical fluorescence imaging,” IEEE J. Quantum Electron. QE23, 1798–1805 (1987). [CrossRef]
  2. W. A. Mohler, J. G. White, “Multiphoton laser scanning microscopy for four-dimensional analysis of caenorhabditis elegans embryonic development,” Opt. Exp. 3, 325–331 (1998); http://www.opticsexpress.org . [CrossRef]
  3. W. J. Denk, J. H. Strickler, W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990). [CrossRef] [PubMed]
  4. M. Gu, “Resolution in three-photon fluorescence scanning microscopy,” Opt. Lett. 21, 988–990 (1996). [CrossRef] [PubMed]
  5. I. Gryczynski, H. Malak, J. R. Lakowicz, “Three-photon induced fluorescence of 2,5-diphenyloxazole with a femetosecond Ti-sapphire laser,” Chem. Phys. Lett. 245, 30–35 (1995). [CrossRef]
  6. I. Gryczynski, H. Malak, J. R. Lakowicz, “Three-photon excitation of a tryptophan derivative using a fs-Ti-sapphire laser,” Biospectroscopy 2, 9–15 (1996). [CrossRef]
  7. V. E. Centonze, J. G. White, “Multiphoton excitation provides optical sections from deeper within scattering specimens than confocal imaging,” Biophys. J. 75, 2015–2024 (1998). [CrossRef] [PubMed]
  8. W. Denk, K. Svoboda, “Photon upmanship: why multiphoton imaging is more than a gimmick,” Neuron 18, 351–357 (1997). [CrossRef] [PubMed]
  9. D. W. Piston, “Imaging living cells and tissues by two-photon excitation microscopy,” Trends Cell Biol. 9, 66–69 (1999). [CrossRef] [PubMed]
  10. S. Charpak, J. Mertz, E. Beaurepaire, L. Moreaux, K. Delaney, “In vivo two-photon imaging of odor-evoked calcium signals in dentrites of rat mitral cells,” Proc. Natl. Acad. Sci. USA 98, 1230–1234 (2001). [CrossRef]
  11. K. Svoboda, W. Denk, D. Kleinfeld, D. W. Tank, “In vivo dentritic calcium dynamics in neocortical pyramid neurons,” Nature (London) 385, 161–165 (1997). [CrossRef]
  12. F. Helmchen, K. Svoboda, W. Denk, D. W. Tank, “In vivo denfritic calcium dynamics in deep-layer cortical pyramidal neurons,” Nature Neurosci. 2, 989–996 (1999). [CrossRef]
  13. K. Svoboda, F. Helmchen, W. Denk, D. W. Tank, “Spread of dendritic excitation in layer 2/3 pyramidal neurons in rat barrel cortex in vivo,” Nature Neuronsci. 2, 65–73 (1999). [CrossRef]
  14. M. Oheim, E. Beaurepaire, E. Chaigneau, J. Mertz, S. Charpak, “Two-photon microscopy in brain tissue: parameters influencing the imaging depth,” J. Neurosci. Methods 111, 29–37 (2001). [CrossRef] [PubMed]
  15. H. J. Schwarzmaier, A. Yaroslavsky, I. Yaroslavsky, T. Goldbach, T. Kahn, F. Ulrich, P. C. Schulze, R. Schober, “Optical properties of native and coagulated human brain structures,” in Lasers in Surgery: Advanced Characterization, Therapeutics, and System VII, R. Anderson, K. E. Bartels, L. S. Bass, K. W. Gregory, D. M. Harris, H. Lui, R. S. Malek, G. T. Mueller, M. M. Pankratov, A. P. Perlmutter, H. Reidenback, L. P. Tate, G. W. Watson, eds., Proc. SPIE2970, 492–499 (1997). [CrossRef]
  16. P. Lenz, “Fluorescence measurement in thick tissue layers by linear or nonlinear long-wavelength excitation,” Appl. Opt. 38, 3662–3669 (1999). [CrossRef]
  17. X. Y. Deng, X. S. Gan, M. Gu, “Multiphoton fluorescence microscopic imaging through double-layer turbid tissue media,” J. Appl. Phys. 91, 4659–4665 (2002). [CrossRef]
  18. X. S. Gan, M. Gu, “Effective point-spread function for fast image modeling and processing in microscopic imaging through turbid media,” Opt. Lett. 24, 741–743 (1999). [CrossRef]
  19. X. S. Gan, M. Gu, “Fluorescence microscope imaging through tissue-like turbid media,” J. Appl. Phys. 87, 3214–3221 (2000). [CrossRef]
  20. M. Gu, X. S. Gan, A. Kisteman, M. Xu, “Comparison of penetration depth between single-photon excitation and two-photon excitation in imaging through turbid tissue media,” Appl. Phys. Lett. 77, 1551–1553 (2000). [CrossRef]
  21. X. S. Gan, M. Gu, “Microscopic image reconstruction through tissue-like turbid media,” Opt. Commun. 207, 149–154 (2002). [CrossRef]
  22. B. Fischl, A. M. Dale, “Measuring the thickness of the human cerebral cortex from magnetic resonance images,” Proc. Natl. Acad. Sci. USA 97, 11044–11049 (2000). [CrossRef]
  23. A. K. Dunn, V. P. Wallace, M. Coleno, M. W. Berns, B. J. Tromberg, “Influence of optical properties on two-photon fluorescence imaging in turbid samples,” App. Opt. 39, 1194–1201 (2000). [CrossRef]
  24. V. Daria, C. M. Blanca, O. Nakamura, S. Kawata, C. Saloma, “Image contrast enhancement for two-photon fluorescence microscopy in a turbid medium,” Appl. Opt. 37, 7960–7967 (1998). [CrossRef]
  25. D. Kleinfeld, P. P. Mitra, F. Helmchen, W. Denk, “Fluctuation and stimulus-induced changes in blood flow observed in individual capillaries in layer 2 through 4 of rat neocortex,” Proc. Natl. Acad. Sci. USA 95, 15741–15746 (1998). [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

OSA is a member of CrossRef.

CrossCheck Deposited