Comparison of penetration depth between two-photon excitation and single-photon excitation in imaging through turbid tissue media

Gu, Miṅ; Gan, Xiaosong; Kisteman, Aernout; Xu, Man

doi:10.1063/1.1308059

Cited by 52 publications

(47 citation statements)

References 12 publications

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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%

“…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%

See 1 more Smart Citation

From molecular structure to tissue architecture: collagen organization probed by SHG microscopy

Cicchi

Vogler

Kapsokalyvas

et al. 2012

Journal of Biophotonics

170

132

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Second‐harmonic generation (SHG) microscopy is a fantastic tool for imaging collagen and probing its hierarchical organization from molecular scale up to tissue architectural level. In fact, SHG combines the advantages of a non‐linear microscopy approach with a coherent modality able to probe molecular organization. In this manuscript we review the physical concepts describing SHG from collagen, highlighting how this optical process allows to probe structures ranging from molecular sizes to tissue architecture, through image pattern analysis and scoring methods. Starting from the description of the most relevant approaches employing SHG polarization anisotropy and forward – backward SHG detection, we then focus on the most relevant methods for imaging and characterizing collagen organization in tissues through image pattern analysis methods, highlighting advantages and limitations of the methods applied to tissue imaging and to potential clinical applications. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

From molecular structure to tissue architecture: collagen organization probed by SHG microscopy

Cicchi

Vogler

Kapsokalyvas

et al. 2012

Journal of Biophotonics

170

132

View full text Add to dashboard Cite

show abstract

“…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%

In vivo monitoring of multiple circulating cell populations using two-photon flow cytometry

Tkaczyk

Zhong

et al. 2008

Optics Communications

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To detect and quantify multiple distinct populations of cells circulating simultaneously in the blood of living animals, we developed a novel optical system for two-channel, two-photon flow cytometry in vivo. We used this system to investigate the circulation dynamics in live animals of breast cancer cells with low (MCF-7) and high (MDA-MB-435) metastatic potential, showing for the first time that two different populations of circulating cells can be quantified simultaneously in the vasculature of a single live mouse. We also non-invasively monitored a population of labeled, circulating red blood cells for more than two weeks, demonstrating that this technique can also quantify the dynamics of abundant cells in the vascular system for prolonged periods of time. These data are the first in vivo application of multichannel flow cytometry utilizing two-photon excitation, which will greatly enhance our capability to study circulating cells in cancer and other disease processes.

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“…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%

Super-resolution endoscopy for real-time wide-field imaging

et al. 2015

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Abstract:Resolving subcellular structures in vitro beyond optical diffraction barrier by a light microscope has achieved significant development since the advancement of super-resolution fluorescence microscopes, such as stimulated emission depletion (STED) microscopy, stochastic optical reconstruction microscopy (STORM) and photoactivated localization microscopy (PALM). However, the resolution of observation in deep and dense in vivo tissues is still confined to cellular level presently, and hence, exploring image details at subcellular level or even beyond organelle level in vivo has continued to attract much research attention. Currently, endoscopy provides an effective way to achieve in vivo observations and is compatible with mature optical microscopy technologies, but its resolution is usually confined to ~1 µm. Here we report a new endoscopy method by functionalizing graded-index (GRIN) lens with microspheres for real-time white-light or fluorescent super-resolution imaging. The capability of resolving objects with feature size of ~λ/5, which breaks the diffraction barrier of traditional GRIN lens based endoscopes by a factor of two, has been demonstrated by using this superresolution endoscopy method. Further development of such a superresolution endoscopy technique may provide new opportunities for in vivo life sciences studies. Jung, and M. J. Schnitzer, "High-speed, miniaturized fluorescence microscopy in freely moving mice," Nat. Methods 5(11), 935-938 (2008).

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Comparison of penetration depth between two-photon excitation and single-photon excitation in imaging through turbid tissue media

Cited by 52 publications

References 12 publications

From molecular structure to tissue architecture: collagen organization probed by SHG microscopy

From molecular structure to tissue architecture: collagen organization probed by SHG microscopy

In vivo monitoring of multiple circulating cell populations using two-photon flow cytometry

Super-resolution endoscopy for real-time wide-field imaging

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