Depth-resolved structural imaging by third-harmonic generation microscopy

Oron, Dan; Yelin, Dvir; Tal, Eran; Raz, Sefi; Fachima, Rachel; Silberberg, Yaron

doi:10.1016/s1047-8477(03)00125-4

Cited by 98 publications

(79 citation statements)

References 23 publications

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“…Therefore, THG is more versatile compared to SHG because it does not require a specific asymmetry of the structure to be imaged. In tissues, THG excitation occurs predominantly at interfaces that are formed between aqueous interstitial fluids and lipid-rich structures, such as cellular membranes (Aptel et al, 2010;Rehberg et al, 2011), lipid droplets (Débarre et al, 2006;Tserevelakis et al, 2014;Watanabe et al, 2010), or inorganic structures, such as calcified bone or enamel in teeth (Chen et al, 2008;Oron et al, 2004). In addition, THG can also be found at interfaces between water and large protein aggregates, such as collagen bundles or muscle fibers (Rehberg et al, 2011;Weigelin et al, 2012).…”

Section: Physical Principles Of Thgmentioning

confidence: 99%

“…1E, examples 5 and 6). Lastly, THG signals are sensitive to the polarization of the excitation light (Oron et al, 2004); in human cornea, using a circularly polarized excitation beam allows the specific detection of interfaces of the lamellae, whereas a linear polarized excitation beam reveals stromal keratocytes and epithelial cell boundaries (Aptel et al, 2010). Thus, signal intensity strongly depends on local imaging conditions.…”

Section: Technical and Experimental Considerations Of Thg Applicationmentioning

confidence: 99%

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Third harmonic generation microscopy of cells and tissue organization

Weigelin

Bakker

Friedl

2016

Journal of Cell Science

179

147

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The interaction of cells within their microenvironmental niche is fundamental to cell migration, positioning, growth and differentiation in order to form and maintain complex tissue organization and function. Third harmonic generation (THG) microscopy is a label-free scatter process that is elicited by water-lipid and water-protein interfaces, including intra-and extracellular membranes, and extracellular matrix structures. In applied life sciences, THG delivers a versatile contrast modality to complement multi-parameter fluorescence, second harmonic generation and fluorescence lifetime microscopy, which allows detection of cellular and molecular cell functions in threedimensional tissue culture and small animals. In this Commentary, we review the physical and technical basis of THG, and provide considerations for optimal excitation, detection and interpretation of THG signals. We further provide an overview on how THG has versatile applications in cell and tissue research, with a particular focus on analyzing tissue morphogenesis and homeostasis, immune cell function and cancer research, as well as the emerging applicability of THG in clinical practice.

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Section: Physical Principles Of Thgmentioning

confidence: 99%

Section: Technical and Experimental Considerations Of Thg Applicationmentioning

confidence: 99%

Third harmonic generation microscopy of cells and tissue organization

Weigelin

Bakker

Friedl

2016

Journal of Cell Science

179

147

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“…Finally, with the use of the same laser, yet at much lower pulse energy, so that no further modification is made to the material, we measure the displacement of the lever amplification beam (Figure 1d) using the third harmonic optical signal generated while scanning the beam across the specimen's surface [43][44][45][46][47][48][49]. …”

Section: Micromachines 2015 6 1368mentioning

confidence: 99%

“…It is a surface-enhanced phenomenon due to its dependence on the different susceptibilities and/or refractive indices of the materials that form an interface. Its nature allows depth-resolved imaging and for this reason its use has been extensively reported for imaging and for obtaining structural information of a wide range of transparent materials from glass and laser-modified glass to biological specimens [43][44][45][46][47][48][49]. Here, by taking advantage of the sensitivity of the THG process to interfaces, we adopt it for in situ metrology.…”

Section: Third-harmonics Generation (Thg) As An In Situ Metrology Toolmentioning

confidence: 99%

A Monolithic Micro-Tensile Tester for Investigating Silicon Dioxide Polymorph Micromechanics, Fabricated and Operated Using a Femtosecond Laser

Athanasiou

Bellouard

2015

Micromachines

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Mechanical testing of materials at the microscales is challenging. It requires delicate procedures not only for producing and handling the specimen to be tested, but also for applying an accurate and controlled force. This endeavor is even more challenging when it comes to investigating the behavior of brittle materials such as glass. Here, we present a microtensile tester for investigating silica glass polymorphs. The instrument is entirely made of silica and for which the same femtosecond laser is not only used for fabricating the device, but also for operating it (loading the specimen) as well as for performing in situ measurements. As a proof-of-concept, we present a stress-strain curve of fused silica for unprecedented high tensile stress of 2.4 GPa, as well as preliminary results of the elastic modulus of femtosecond laser-affected zones of fused silica, providing new insights on their microstructures and mechanical behavior.

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“…In this article, we will focus on third-harmonic generation (THG) microscopy [14,15], an imaging modality which has proven useful for biological imaging [4,[16][17][18][19] and which is particularly sensitive to the interplay between sample and field structure. The most remarkable characteristic of THG microscopy with a Gaussian excitation beam is that no signal is obtained from a homogeneous positively dispersive sample [14].…”

Section: Introductionmentioning

confidence: 99%

Third-harmonic generation microscopy with Bessel beams: a numerical study

Olivier¹,

Débarre²,

Mahou³

et al. 2012

Opt. Express

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Abstract:We study theoretically and numerically third-harmonic generation (THG) from model geometries (interfaces, slabs, periodic media) illuminated by Bessel beams produced by focusing an annular intensity profile. Bessel beams exhibit a phase and intensity distribution near focus different from Gaussian beams, resulting in distinct THG phase matching properties and coherent scattering directions. Excitation wave vectors are controlled by adjusting the bounding aperture angles of the Bessel beam. In addition to extended depth-of-field imaging, this opens interesting perspectives for coherent nonlinear microscopy, such as extracting sample spatial frequencies in the λ /8 -λ range in the case of organized media.

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Depth-resolved structural imaging by third-harmonic generation microscopy

Cited by 98 publications

References 23 publications

Third harmonic generation microscopy of cells and tissue organization

Third harmonic generation microscopy of cells and tissue organization

A Monolithic Micro-Tensile Tester for Investigating Silicon Dioxide Polymorph Micromechanics, Fabricated and Operated Using a Femtosecond Laser

Third-harmonic generation microscopy with Bessel beams: a numerical study

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