Second Harmonic Generation Emission Research Articles

Enhancement of the Second Harmonic Generation from WS2 Monolayers by Cooperating with Dielectric Microspheres

AbstractHerein, a 20‐fold enhancement of the optical second harmonic generation (SHG) from WS2 monolayers by cooperating with SiO2 dielectric microspheres (MSs) is reported. Detailed experiment and simulation studies indicate that the SHG enhancement is dominantly originated by the enhanced excitation intensity and enlarged collection efficiency produced by the SiO2 MSs. The size effect of the SiO2 MS on the SHG enhancement is also systematically investigated. Moreover, the SHG enhancement is found to be very sensitive to the surrounding refractive index of the SiO2 MS, which advocates great promise for the photonic sensing applications. This work provides a low‐cost and efficient approach to enhancing the SHG emission, and demonstrates great potential for functional nanodevices based on hybrid dielectric structures.

Nonlinear Optics: Enhanced Second Harmonic Generation in Double‐Resonance Colloidal Metasurfaces (Adv. Funct. Mater. 51/2018)

In article number 1803019, Andrea R. Tao and co-workers describe a new protocol to fabricate double resonance Second Harmonic Generation (SHG) metasurfaces in wafer scale. Langmuir-Blodgett films, obtained by dropping silver nanocubes (AgNCs) on an air-water interface, are transferred to a silica-supported Au substrate to form a metasurface in wafer scale. Near-Infrared laser (900 nm) excitation causes a strong SHG emission at 450 nm, emitted from the nanometer scale gap between AgNCs and Au substrate.

Directional second-harmonic generation controlled by sub-wavelength facets of an organic mesowire.

Directional harmonic generation is an important property characterizing the ability of nonlinear optical antennas to diffuse the signal in a well-defined region of space. Herein, we show how sub-wavelength facets of an organic molecular mesowire crystal can be utilized to systematically vary the directionality of second-harmonic generation (SHG) in the forward-scattering geometry. We demonstrate this capability on crystalline diamonoanthraquinone (DAAQ) mesowires with sub-wavelength facets. We observed that the radial angles of the SHG emission can be tuned over a range of 130deg. This angular variation arises due to spatially distributed nonlinear dipoles in the focal volume of the excitation as well as the geometrical cross section and facet orientation of the mesowire. Numerical simulations of the near-field excitation profile corroborate the role of the mesowire geometry in localizing the electric field. In addition to directional SHG from the mesowire, we experimentally observe optical waveguiding of the nonlinear two-photon excited fluorescence (TPEF). Interestingly, we observed that for a given pump excitation, the TPEF signal is isotropic and delocalized, whereas the SHG emission is directional and localized at the location of excitation. All the observed effects have direct implications not only in active nonlinear optical antennas but also in nonlinear signal processing.

Assessing local stromal alterations in human ovarian cancer subtypes via second harmonic generation microscopy and analysis

.The collagen architecture in all human ovarian cancers is substantially remodeled, where these alterations are manifested in different fiber widths, fiber patterns, and fibril size and packing. Second harmonic generation (SHG) microscopy has differentiated normal tissues from high-grade serous (HGS) tumors with high accuracy; however, the classification between low-grade serous, endometrioid, and benign tumors was less successful. We postulate this is due to known higher genetic variation in these tissues relative to HGS tumors, which are genetically similar, and this results in more heterogeneous collagen remodeling in the respective matrix. Here, we examine fiber widths and SHG emission intensity and directionality locally within images (e.g., ) and show that normal tissues and HGS tumors are more uniform in fiber properties as well as in fibril size and packing than the other tissues. Moreover, these distributions are in good agreement with phase matching considerations relating SHG emission directionality and intensity. The findings show that in addition to average collagen assembly properties the intrinsic heterogeneity must also be considered as another aspect of characterization. These local analyses showed differences not shown in pure intensity-based image analyses and may provide further insight into disease etiology of the different tumor subtypes.

Efficient continuous-wave nonlinear frequency conversion in high-Q gallium nitride photonic crystal cavities on silicon

We report on nonlinear frequency conversion from the telecom range via second harmonic generation (SHG) and third harmonic generation (THG) in suspended gallium nitride slab photonic crystal (PhC) cavities on silicon, under continuous-wave resonant excitation. Optimized two-dimensional PhC cavities with augmented far-field coupling have been characterized with quality factors as high as 4.4 × 104, approaching the computed theoretical values. The strong enhancement in light confinement has enabled efficient SHG, achieving a normalized conversion efficiency of 2.4 × 10−3 W−1, as well as simultaneous THG. SHG emission power of up to 0.74 nW has been detected without saturation. The results herein validate the suitability of gallium nitride for integrated nonlinear optical processing.

Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections.

Nonlinear microscopy, namely multiphoton imaging and second harmonic generation (SHG), is an established noninvasive technique useful for the imaging of extracellular matrix (ECM). Typically, measurements are performed in vivo on freshly excised tissues or biopsies. In this article, we describe the effect of rehydrating paraffin-embedded sections on multiphoton and SHG emission signals and the acquisition of nonlinear images from hematoxylin and eosin (H&E)-stained sections before and after a destaining protocol. Our results reveal that bringing tissue sections to a physiological state yields a significant improvement in nonlinear signals, particularly in SHG. Additionally, the destaining of sections previously processed with H&E staining significantly improves their SHG emission signals during imaging, thereby allowing sufficient analysis of collagen in these sections. These results are important for researchers and pathologists to obtain additional information from paraffin-embedded tissues and archived samples to perform retrospective analysis of the ECM or gain additional information from rare samples.

Quantitative structural markers of colorectal dysplasia in a cross sectional study of ex vivo murine tissue using label-free multiphoton microscopy.

Two-photon excitation of label-free tissue is of increasing interest, as advances have been made in endoscopic clinical application of multiphoton microscopy, such as second harmonic generation (SHG) scanning endoscopy used to monitor cervical collagen in mice1. We used C57BL mice as a model to investigate the progression of gastrointestinal structures, specifically glandular area and circularity. We used multiphoton microscopy to image ex-vivo label-free murine colon, focusing on the collagen structure changes over time, in mice ranging from 10 to 20 weeks of age. Series of images were acquired within the colonic and intestinal tissue at depth intervals of 20 microns from muscularis to the epithelium, up to a maximum depth of 180 microns. The imaging system comprised a two-photon laser tuned to 800nm wavelength excitation, and the SHG emission was filtered with a 400/40 bandpass filter before reaching the photomultiplier tube. Images were acquired at 15 frames per second, for 200 to 300 cumulative frames, with a field of view of 261um by 261um, and 40mW at sample. Image series were compared to histopathology H&E slides taken from adjacent locations. Quantitative metrics for determining differences between murine glandular structures were applied, specifically glandular area and circularity.

Single potassium niobate nano/microsized particles as local mechano-optical Brownian probes.

Perovskite alkaline niobates, due to their strong nonlinear optical properties, including birefringence and the capability to produce second-harmonic generation (SHG) signals, attract a lot of attention as potential candidates for applications as local nano/microsized mechano-optical probes. Here, we report on an implementation of photonic force microscopy (PFM) to explore the Brownian motion and optical trappability of monocrystalline potassium niobate (KNbO3) nano/microsized particles having sizes within the range of 50 to 750 nm. In particular, we exploit the anisotropic translational diffusive regime of the Brownian motion to quantify thermal fluctuations and optical forces of singly-trapped KNbO3 particles within the optical trapping volume of a PFM microscope. We also show that, under near-infrared (NIR) excitation of the highly focused laser beam of the PFM microscope, a single optically-trapped KNbO3 particle reveals a strong SHG signal manifested by a narrow peak (λ(em) = 532 nm) at half the excitation wavelength (λ(ex) = 1064 nm). Moreover, we demonstrate that the thus induced SHG emission can be used as a local light source that is capable of optically exciting molecules of an organic dye, Rose Bengal (RB), which adhere to the particle surface, through the mechanism of luminescence energy transfer (LET).

Phase matching of backward second harmonic generation assisted by lattice structure in collagen tissues.

Phase matching of backward second harmonic generation (SHG) in a periodic structure of collagen fibrils is investigated through theoretical modeling, simulation, and experiments. The lattice structure of collagen fibrils is considered to provide a virtual momentum for assisting the phase matching of backward SHG. Phase matching over a relatively wide excitation wavelength range is achieved by tilting the angle of the fundamental excitation and SHG wave vectors. The SHG intensity in the periodic structure is simulated to quantify the phase matching effect. The effect of the fundamental excitation and the SHG emission angles on the peak excitation wavelength of the SHG excitation spectrum is further validated in experiments, where the excitation and emission angles are controlled by spatial filtering. It is found that an optimized excitation wavelength exists for a certain collagen fibril structure, which shifts toward a shorter wavelength when the excitation and emission angles are increased. Our results show that the lattice structure of collagen fibrils can assist the phase matching, providing a mechanism for generating backward SHG in multiphoton microscopy.

Single-cell screening and quantification of transcripts in cancer tissues by second-harmonic generation microscopy.

Fluorescence-based single molecule techniques to interrogate gene expression in tissues present a very low signal-to-noise ratio due to the strong autofluorescence and other background signals from tissue sections. This report presents a background-free method using second-harmonic generation (SHG) nanocrystals as probes to quantify the messenger RNA (mRNA) of human epidermal growth receptor 2 (Her2) at single molecule resolution in specific phenotypes at single-cell resolution directly in tissues. Coherent SHG emission from individual barium titanium oxide (BTO) nanoprobes was demonstrated, allowing for a stable signal beyond the autofluorescence window. Her2 surface marker and Her2 mRNA were specifically labeled with BTO probes, and Her2 mRNA was quantified at single copy sensitivity in Her2 expressing phenotypes directly in cancer tissues. Our approach provides the first proof of concept of a cross-platform strategy to probe tissues at single-cell resolution in situ.

Second-Harmonic Generation from Sub-5 nm Gaps by Directed Self-Assembly of Nanoparticles onto Template-Stripped Gold Substrates.

Strong field enhancement and confinement in plasmonic nanostructures provide suitable conditions for nonlinear optics in ultracompact dimensions. Despite these enhancements, second-harmonic generation (SHG) is still inefficient due to the centrosymmetric crystal structure of the bulk metals used, e.g., Au and Ag. Taking advantage of symmetry breaking at the metal surface, one could greatly enhance SHG by engineering these metal surfaces in regions where the strong electric fields are localized. Here, we combine top-down lithography and bottom-up self-assembly to lodge single rows of 8 nm diameter Au nanoparticles into trenches in a Au film. The resultant "double gap" structures increase the surface-to-volume ratio of Au colocated with the strong fields in ∼2 nm gaps to fully exploit the surface SHG of Au. Compared to a densely packed arrangement of AuNPs on a smooth Au film, the double gaps enhance SHG emission by 4200-fold to achieve an effective second-order susceptibility χ((2)) of 6.1 pm/V, making it comparable with typical nonlinear crystals. This patterning approach also allows for the scalable fabrication of smooth gold surfaces with sub-5 nm gaps and presents opportunities for optical frequency up-conversion in applications that require extreme miniaturization.

Articular cartilage zonal differentiation via 3D Second-Harmonic Generation imaging microscopy

Purpose: The collagen structure throughout the patella has not been thoroughly investigated by 3D imaging, where the majority of the existing data come from histological cross sections. It is important to have a better understanding of the architecture in normal tissues, where this could then be applied to imaging of diseased states.Methods: To address this shortcoming, we investigated the combined use of collagen-specific Second-Harmonic Generation (SHG) imaging and measurement of bulk optical properties to characterize collagen fiber orientations of the histologically defined zones of bovine articular cartilage. Forward and backward SHG intensities of sections from superficial, middle and deep zones were collected as a function of depth and analyzed by Monte Carlo simulations to extract the SHG creation direction, which is related to the fibrillar assembly.Results: Our results revealed differences in SHG forward–backward response between the three zones, where these are consistent with a previously developed model of SHG emission. Some of the findings are consistent with that from other modalities; however, SHG analysis showed the middle zone had the most organized fibril assembly. While not distinct, we also report bulk optical property values for these different zones within the patella.Conclusions: Collectively, these results provide quantitative measurements of structural changes at both the fiber and fibril assembly of the different cartilage zones and reveals structural information not possible by other microscope modalities. This can provide quantitative insight to the collagen fiber network in normal cartilage, which may ultimately be developed as a biomarker for osteoarthritis.

Eu-doping induced improvement on the second harmonic generation of ZnO Nanowires

ABSTRACTWe report the Eu doping induced improvement on the second harmonic generation (SHG) of ZnO nanowires and correlates with the structural modification and corresponding linear absorption. A non-monotonic enhancement in the SHG emission is observed with the increase of Eu concentration. To understand the underlying mechanism, the effective second order non–linear coefficient (deff) is calculated from the theoretical fitting with considering the absorption effect. The highest deff (19.09±0.11 pm/V) is obtained for the 1 at.% Eu doped ZnO nanowires, which is several times larger than the standard SHG material β-BaB2O4 (BBO). Dependence of the deff with the Eu doping, structural modification and absorption magnitude are systematically discussed.

Simultaneous determination of the second-harmonic generation emission directionality and reduced scattering coefficient from three-dimensional imaging of thick tissues

Second-harmonic generation (SHG) microscopy has intrinsic contrast for imaging fibrillar collagen and has shown great promise for disease characterization and diagnostics. In addition to morphology, additional information is achievable as the initially emitted SHG radiation directionality is related to subresolution fibril size and distribution. We show that by two parameter fittings, both the emission pattern (FSHG/BSHG)creation and the reduced scattering coefficient μs', can be obtained from the best fits between three-dimensional experimental data and Monte Carlo simulations. The improved simulation framework accounts for collection apertures for the detected forward and backward components. We apply the new simulation framework to mouse tail tendon for validation and show that the spectral slope of μs' obtained is similar to that from bulk optical measurements and that the (FSHG/BSHG)creation values are also similar to previous results. Additionally, we find that the SHG emission becomes increasingly forward directed at longer wavelengths, which is consistent with decreased dispersion in refractive index between the laser and SHG wavelengths. As both the spectral slope of μs' and (FSHG/BSHG)creation have been linked to the underlying tissue structure, simultaneously obtaining these parameters on a microscope platform from the same tissue provides a powerful method for tissue characterization.

Tumor-associated macrophages and stromal TNF-α regulate collagen structure in a breast tumor model as visualized by second harmonic generation

Collagen fibers can be imaged with second harmonic generation (SHG) and are associated with efficient tumor cell locomotion. Preferential locomotion along these fibers correlates with a more aggressively metastatic phenotype, and changes in SHG emission properties accompany changes in metastatic outcome. We therefore attempted to elucidate the cellular and molecular machinery that influences SHG in order to understand how the microstructure of tumor collagen fibers is regulated. By quantifying SHG and immunofluorescence (IF) from tumors grown in mice with and without stromal tumor necrosis factor (TNF)-α and in the presence or absence of tumor-associated macrophages (TAMs), we determined that depletion of TAMs alters tumor collagen fibrillar microstructure as quantified by SHG and IF. Furthermore, we determined that abrogation of TNF-α expression by tumor stromal cells also alters fibrillar microstructure and that subsequent depletion of TAMs has no further effect. In each case, metastatic burden correlated with optical readouts of collagen microstructure. Our results implicate TAMs and stromal TNF-α as regulators of breast tumor collagen microstructure and suggest that this regulation plays a role in tumor metastasis. Furthermore, these results indicate that quantification of SHG represents a useful strategy for evaluating the cells and molecular pathways responsible for manipulating fibrillar collagen in breast tumor models.

Second harmonic generation signal in collagen fibers: role of polarization, numerical aperture, and wavelength

The spatial distribution of second harmonic generation (SHG) signal from collagen fibers for incident elliptical polarized light has been modeled. The beam was assumed to focus on a horizontal fiber through a microscope objective. For elliptical polarized states located along a vertical meridian of the Poincare sphere, the SHG intensity has been optimized in terms of the incident wavelength and the numerical aperture (NA) of the objective. Our results show that polarization modulates the SHG signal. Elliptical polarization can generate high signals (even greater than those corresponding to linear polarization) when combined with appropriate values of both incident wavelength and NA. On the other hand, the SHG intensity might also be identically zero for particular elliptical polarization states, a condition that depends exclusively on the ratio of hyperpolarizabilities of the collagen fibers. The highest forward-to-backward SHG signal distribution occurs along the propagating direction, depends on the incident wavelength, and reduces when NA increases. Furthermore, the direction of maximum SHG emission was found to be more sensitive to changes in the NA rather than variations in the incident wavelength. These findings could help to optimize the experimental conditions of multiphoton microscopes and to increase SHG signals from biological tissues containing collagen.

Structural changes in mixed Col I/Col V collagen gels probed by SHG microscopy: implications for probing stromal alterations in human breast cancer

Second Harmonic Generation (SHG) microscopy has been previously used to describe the morphology of collagen in the extracellular matrix (ECM) in different stages of invasion in breast cancer. Here this concept is extended by using SHG to provide quantitative discrimination of self-assembled collagen gels, consisting of mixtures of type I (Col I) and type V (Col V) isoforms which serve as models of changes in the ECM during invasion in vivo. To investigate if SHG is sensitive to changes due to Col V incorporation into Col I fibrils, gels were prepared with 0-20% Col V with the balance consisting of Col I. Using the metrics of SHG intensity, fiber length, emission directionality, and depth-dependent intensities, we found similar responses for gels comprised of 100% Col I, and 95% Col I/5% Col V, where these metrics were all significantly different from those of the 80% Col I/20% Col V gels. Specifically, the gels of lower Col V content produce brighter SHG, are characterized by longer fibers, and have a higher forward/backward emission ratio. These attributes are all consistent with more highly organized collagen fibrils/fibers and are in agreement with previous TEM characterization as well as predictions based on phase matching considerations. These results suggest that SHG can be developed to discriminate Col I/Col V composition in tissues to characterize and follow breast cancer invasion.

The origin of second harmonic generation hotspots in chiral optical metamaterials [Invited

Novel ways to detect the handedness in chiral optical metamaterials by means of the second harmonic generation (SHG) process have recently been proposed. However, the precise origin of the SHG emission has yet to be unambiguously established. In this paper, we present computational simulations of both the electric currents and the electromagnetic fields in chiral planar metamaterials, at the fundamental frequency (FF), and discuss the implications of our results on the characteristics of experimentally measured SHG. In particular, we show that the results of our numerical simulations are in good agreement with the experimental mapping of SHG sources. Thus, the SHG in these metamaterials can be attributed to a strong local enhancement of the electromagnetic fields at the FF, which depends on the particular structure of the patterned metamaterial.

Backward emission angle of microscopic second-harmonic generation from crystallized type I collagen fiber

A theoretical model that deals with SHG from crystallized type I collagen fiber formed by a bundle of fibrils is established. By introducing a density distribution function of dipoles within fibrils assembly into the dipole theory and combining with structural order (m,l) parameters revealed by quasi-phase-matching (QPM) theory, our established theoretical model comprehensively characterizes both biophysical features of collagen dipoles and the crystalline characteristics of collagen fiber. This new model quantitatively reveals the 3-D distribution of second-harmonic generation (SHG) emission angle (θ,ϕ) in accordance with the emission power. Results show that fibrils diameter d(1) and structural order m, which describes the structural characteristics of collagen fiber along the incident light propagation direction has significant influence on backward∕forward SHG emission. The decrease of fibrils diameter d(1) induces an increase of the peak SHG emission angle θ(max). As d(1) decreases to a threshold value, in our case it is around d(1) = 150 nm when (m,l) = (1,0), θ(max) > 90 deg, indicating that backward SHG emission appears. The SHG may have two symmetrical emission distribution lobes or may have only one or two unsymmetrical emission lobes with unequal emission power, depending on the functional area of (m,l) on d(1).

Microscopic second harmonic generation (SHG) from tilt-placed collagen fibrils

We report a theoretical study of second-harmonic generation (SHG) from the collagen fibrils which have a tilt angle Φ between their axes with the plane of the polarization angle α of the linearly polarized focused light. The effects of Φ as well as α on SHG emission have been investigated. Our results show that the total strength of SHG electrical field (E(2ω)) and power (P(2ω)) reach their maximal values as Φ locates at a certain angle, here it is around 27° under all demonstrated α. When Φ < 27°, E(2ω) and P(2ω) gradually increase to their maximal value, while when Φ > 27°, they quickly drops. Specially, E(2ω) and P(2ω) are almost undetectable when Φ ≥ 45°. Also, Φ influences the distribution pattern of E(2ω), P(2ω) and their parallel (E(2ω,p), P(2ω,p)) and perpendicular (E(2ω,s), P(2ω,s)) components, such as the number of emission lobes and shape. α has unevenly impact on E(2ω) and P(2ω), the far deviation of α from x axis induce much great decrease of E(2ω) and P(2ω). α = 45° has special influence on the distribution pattern of E(2ω,s), E(2ω), P(2ω,s) as well as P(2ω) compared to those of α = 0° and 90°.