Spatially distributed two-photon excitation fluorescence in scattering media: Experiments and time-resolved Monte Carlo simulations - Archive ouverte HAL
Article Dans Une Revue Optics Communications Année : 2007

Spatially distributed two-photon excitation fluorescence in scattering media: Experiments and time-resolved Monte Carlo simulations

A. Leray
Christophe Odin
E. Huguet
  • Fonction : Auteur
François Amblard
  • Fonction : Auteur
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Résumé

We report on measurements and Monte Carlo analysis of the spatial distribution of light in two-photon excitation fluorescence (TPEF) microscopy operating through scattering media. The axial profile of TPEF produced by a high numerical aperture microscope objective focusing through a tissue-like optical phantom at depths more than five photon mean-scattering lengths is studied. The measured profile is quantitatively interpreted with time-resolved Monte Carlo simulations that take into account the spatio-temporal distribution of the photons within the turbid medium under a femtosecond excitation regime. It is shown that considering only the ballistic photons leads to an under-estimation of the actual out-of-focus distribution of the fluorescence, whereas it is over-estimated when the ballistic and scattered photons are treated alike. Comparison of the in-focus signal to the overall out-of-focus background clearly pointed out that the signal-to-background ratio of TPEF microscopy images benefits from a shortening of the excitation pulses. Moreover, in homogeneously labeled specimen, the background is shown to overcome the signal at imaging depths of about five to six photon mean-scattering paths.

Dates et versions

hal-00907971 , version 1 (22-11-2013)

Identifiants

Citer

A. Leray, Christophe Odin, E. Huguet, François Amblard, Yann Le Grand. Spatially distributed two-photon excitation fluorescence in scattering media: Experiments and time-resolved Monte Carlo simulations. Optics Communications, 2007, 272 (1), pp.269-278. ⟨10.1016/J.OPTCOM.2006.11.007⟩. ⟨hal-00907971⟩
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