Spectroscopic analysis of keratin endogenous signal for skin multiphoton microscopy

Pena, Ana‐Maria; Strupler, Mathias; Boulesteix, Thierry; Schanne-Klein, Marie-Claire

doi:10.1364/opex.13.006268

Cited by 136 publications

(122 citation statements)

References 21 publications

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“…Epithelial cells 277/382 nm (Pena et al, 2005). 700-900/460-550 nm (760/475 nm; 860/515 nm) (Pena et al, 2005;Wu & Qu, 2005 Georgakoudi et al, 2002).…”

Section: Keratinmentioning

confidence: 99%

Two‐photon microscopy of deep intravital tissues and its merits in clinical research

Wang

König

2010

Journal of Microscopy

180

131

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SummaryMultiphoton excitation laser scanning microscopy, relying on the simultaneous absorption of two or more photons by a molecule, is one of the most exciting recent developments in biomedical imaging. Thanks to its superior imaging capability of deeper tissue penetration and efficient light detection, this system becomes more and more an inspiring tool for intravital bulk tissue imaging. Two-photon excitation microscopy including 2-photon fluorescence and second harmonic generated signal microscopy is the most common multiphoton microscopic application. In the present review we take diverse ocular tissues as intravital samples to demonstrate the advantages of this approach. Experiments with registration of intracellular 2-photon fluorescence and extracellular collagen second harmonic generated signal microscopy in native ocular tissues are focused. Data show that the in-tandem combination of 2-photon fluorescence and second harmonic generated signal microscopy as two-modality microscopy allows for in situ co-localization imaging of various microstructural components in the whole-mount deep intravital tissues. New applications and recent developments of this high technology in clinical studies such as 2-photon-controlled drug release, in vivo drug screening and administration in skin and kidney, as well as its uses in tumourous tissues such as melanoma and glioma, in diseased lung, brain and heart are additionally reviewed. Intrinsic emission two-modal 2-photon microscopy/tomography, acting as an efficient and sensitive non-injurious imaging approach featured by high contrast and subcellular spatial resolution, has been proved to be a promising tool for intravital deep tissue imaging and clinical studies. Given the level of its performance, we believe Correspondence to B.-G. Wang. Tel: +49 3641 938496; fax: +49 3641 938552; e-mail: Baogui.Wang@mti.uni-jena.de that the non-linear optical imaging technique has tremendous potentials to find more applications in biomedical fundamental and clinical research in the near future.

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“…Epithelial cells 277/382 nm (Pena et al, 2005). 700-900/460-550 nm (760/475 nm; 860/515 nm) (Pena et al, 2005;Wu & Qu, 2005 Georgakoudi et al, 2002).…”

Section: Keratinmentioning

confidence: 99%

Two‐photon microscopy of deep intravital tissues and its merits in clinical research

Wang

König

2010

Journal of Microscopy

180

131

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“…A large number of studies using TPEF on tissues have been conducted ex vivo [18] and also in vivo, on both living animals or humans [19][20][21][22] demonstrating the capabilities of this technique for label-free morphological imaging in deep tissues. Functional information can be provided by optical separation of molecular autofluorescence signals using time-resolved (FLIM) [23][24][25][26][27][28][29][30], spectral-resolved approaches [31][32][33][34] or both in tandem [35][36][37]. A complementary NLO imaging technique is coherent anti-Stokes Raman scattering (CARS) microscopy [38][39][40], which provides high-resolution images of the symmetric and asymmetric CH-stretching vibration [41][42][43], i.e.…”

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

171

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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“…The cortex of the human hair is birefringent, where the ordinary and extraordinary refractive indices, n 0 and n E , are 1.541 and 1.548, respectively and the corresponding value of birefringence, d, is equal to 0.007. This anisotropy of the refractive index is due to the intrinsic and form birefringence [20], resulting both from the a-helix structure of keratin fibers and the refractive index difference between the keratin fibers and the surrounding amorphous matrices, respectively [21]. If the angle between the hair axis and the incoming light polarization is 0 or 90 , the light traversed through the hair experiences no birefringence and consequently no change in polarization (apart from that induced by light reflectance at the hair interface, which is described by the Fresnel equations).…”

Section: Discussionmentioning

confidence: 99%

“…Other methods, such as confocal auto fluorescence imaging, based on the detection of auto fluorescence of keratin [21] and melanin [22] can also be utilized for imaging hairs. On practice this can be realized by using one-and two photon-excited auto fluorescence processes [23].…”

Section: Discussionmentioning

confidence: 99%

Contrast improvement in scattered light confocal imaging of skin birefringent structures by depolarization detection

Varghese

Verhagen

Tai

et al. 2011

Journal of Biophotonics

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Journal of BIOPHOTONICSHere we describe a method for enhancing the contrast in imaging skin birefringent structures. The method relies on polarization-dependent optical properties and is implemented using cross polarized confocal microscopy. The experimental data obtained using ex-vivo and invivo measurements on human scalp hairs and human skin demonstrate a significant dependence of the change in polarization of light that interacted with the birefringent hair on the orientation of the incident polarization. The polarization dependent contrast, defined as the ratio of intensity measured for different orientations of the incident polarization when observed using cross polarized confocal microscopy furthermore depends on the hair type/degree of pigmentation and on the focusing depth inside the hair. No such dependence was observed for the upper skin layers, including the stratum corneum and epidermis. We propose a new method for enhancing the contrast between the skin and the birefringent hair by the use of cross polarized confocal microscopy combined with the variation of the polarization of the incoming light. Potential applications of this method include imaging of hairs for assessing the efficacy of hair removal methods and measurement of skin birefringence. The underestimation of the birefringence content resulting from the orientation related effects associated with the use of linearly polarized light for imaging tissues containing wavy birefringent structures could be minimized by this method.CPCLSM images obtained in-vivo with the direction of incident polarization at different angles to the orientation of the hair axis.

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Spectroscopic analysis of keratin endogenous signal for skin multiphoton microscopy

Cited by 136 publications

References 21 publications

Two‐photon microscopy of deep intravital tissues and its merits in clinical research

Two‐photon microscopy of deep intravital tissues and its merits in clinical research

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

Contrast improvement in scattered light confocal imaging of skin birefringent structures by depolarization detection

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