2000
DOI: 10.1063/1.1308059
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Comparison of penetration depth between two-photon excitation and single-photon excitation in imaging through turbid tissue media

Abstract: We show, both theoretically and experimentally, that for a turbid tissue medium where Mie scattering is dominant, multiple scattering not only reduces the illumination power in the forward direction but also exhibits an anisotropic distribution of scattered photons. Thus, a signal level under two-photon excitation drops much faster than that under single-photon excitation although image resolution is much higher in the former case. As a result, the penetration depth under two-photon excitation is limited by th… Show more

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Cited by 52 publications
(47 citation statements)
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“…Although every biological tissue intrinsically contains a certain amount of fluorescent molecules with excitation bands in the UV-blue region of the spectrum, light penetration into optically turbid samples is strongly limited by scattering and absorption and -as a consequence -imaging is limited to the outermost layers. In NLO microscopy, the use of NIR wavelengths offers the advantage of a reduced scattering with respect to imaging by linear techniques and hence a deeper penetration inside optically turbid samples as biological tissues [7,14]. Furthermore, NLO imaging intrinsically offers a spatial resolution comparable with respect to equivalent linear techniques (i.e.…”
Section: Introductionmentioning
confidence: 99%
See 1 more Smart Citation
“…Although every biological tissue intrinsically contains a certain amount of fluorescent molecules with excitation bands in the UV-blue region of the spectrum, light penetration into optically turbid samples is strongly limited by scattering and absorption and -as a consequence -imaging is limited to the outermost layers. In NLO microscopy, the use of NIR wavelengths offers the advantage of a reduced scattering with respect to imaging by linear techniques and hence a deeper penetration inside optically turbid samples as biological tissues [7,14]. Furthermore, NLO imaging intrinsically offers a spatial resolution comparable with respect to equivalent linear techniques (i.e.…”
Section: Introductionmentioning
confidence: 99%
“…Non-linear optical processes play a crucial role in terms of achievable penetration depth [6][7][8], spatial resolution [9,10], and excitation capability [11][12][13]. Although every biological tissue intrinsically contains a certain amount of fluorescent molecules with excitation bands in the UV-blue region of the spectrum, light penetration into optically turbid samples is strongly limited by scattering and absorption and -as a consequence -imaging is limited to the outermost layers.…”
Section: Introductionmentioning
confidence: 99%
“…Using the simultaneous injection of labeled RBCs as a reference control for focal point optimization will enable far more reliable experiments on rare event circulation dynamics. One solution to increase the statistical power of the measurements would be to access larger blood vessels with higher flow rates, thereby increasing the number of detected cells; this should be enabled by the ability of the femtosecond NIR laser source to penetrate much deeper through biological tissue than a continuous wave laser in visible region for one-photon excitation [18], and to potentially reduced photo damage to surrounding tissue for extended monitoring.…”
Section: Discussionmentioning
confidence: 99%
“…By taking the advantages of penetration ability of nearinfrared light and objectives with high numerical aperture (NA), two-photon fluorescence microscopy enables three-dimensional tissue imaging as deep as 500 -1000 µm [3,4] with resolution of cellular level [3][4][5][6]. In order to extend the penetration depth of two-photon fluorescence microscopy, several strategies have been proposed, such as enhancement of the fluorescence signals generation, improvement of optical signal collection and use of endoscope [2].…”
Section: Introductionmentioning
confidence: 99%