Spatially distributed two-photon excitation fluorescence in scattering media: Experiments and time-resolved Monte Carlo simulations
| Autor: | Aymeric Leray, Etienne Huguet, Y. Le Grand, François Amblard, Christophe Odin |
|---|---|
| Přispěvatelé: | Groupe matière condensée et matériaux (GMCM), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Physico-Chimie-Curie (PCC), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut Curie [Paris]-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC) |
| Rok vydání: | 2007 |
| Předmět: |
Physics
0303 health sciences Photon business.industry Scattering Monte Carlo method 01 natural sciences Atomic and Molecular Physics and Optics Electronic Optical and Magnetic Materials Numerical aperture 010309 optics 03 medical and health sciences Optics Two-photon excitation microscopy 0103 physical sciences Femtosecond Microscopy [PHYS.COND.CM-MS]Physics [physics]/Condensed Matter [cond-mat]/Materials Science [cond-mat.mtrl-sci] Electrical and Electronic Engineering Physical and Theoretical Chemistry Ballistic photon business 030304 developmental biology |
| Zdroj: | Optics Communications Optics Communications, 2007, 272 (1), pp.269-278. ⟨10.1016/J.OPTCOM.2006.11.007⟩ Optics Communications, Elsevier, 2007, 272 (1), pp.269-278. ⟨10.1016/J.OPTCOM.2006.11.007⟩ |
| ISSN: | 0030-4018 |
| DOI: | 10.1016/j.optcom.2006.11.007 |
| Popis: | International audience; 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. |
| Databáze: | OpenAIRE |
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