Measuring the effect of a Western diet on liver tissue architecture by FLIM autofluorescence and harmonic generation microscopy

PubMed Central

Ranjit, Suman; Dvornikov, Alexander; Dobrinskikh, Evgenia; Wang, Xiaoxin; Luo, Yuhuan; Levi, Moshe; Gratton, Enrico

2017-01-01

The phasor approach to auto-fluorescence lifetime imaging was used to identify and characterize a long lifetime species (LLS) (~7.8 ns) in livers of mice fed with a Western diet. The size of the areas containing this LLS species depends on the type of diet and the size distribution shows Western diet has much larger LLS sizes. Combination of third harmonic generation images with FLIM identified the LLS species with fat droplets and the droplet size distribution was estimated. Second harmonic generation microscopy combined with phasor FLIM shows that there is an increase in fibrosis with a Western diet. A new decomposition in three components of the phasor plot shows that a Western diet is correlated with a higher fraction of free NADH, signifying more reducing condition and more glycolytic condition. Multiparametric analysis of phasor distribution shows that from the distribution of phasor points, a Western diet fed versus a low fat diet fed samples of mice livers can be separated. The phasor approach for the analysis of FLIM images of autofluorescence in liver specimens can result in discovery of new fluorescent species and then these new fluorescent species can help assess tissue architecture. Finally integrating FLIM and second and third harmonic analysis provides a measure of the advancement of fibrosis as an effect of diet. PMID:28717559

Third-harmonic generation susceptibility spectroscopy in free fatty acids

NASA Astrophysics Data System (ADS)

Chen, Yu-Cheng; Hsu, Hsun-Chia; Lee, Chien-Ming; Sun, Chi-Kuang

2015-09-01

Lipid-correlated disease such as atherosclerosis has been an important medical research topic for decades. Many new microscopic imaging techniques such as coherent anti-Stokes Raman scattering and third-harmonic generation (THG) microscopy were verified to have the capability to target lipids in vivo. In the case of THG microscopy, biological cell membranes and lipid bodies in cells and tissues have been shown as good sources of contrast with a laser excitation wavelength around 1200 nm. We report the THG excitation spectroscopy study of two pure free fatty acids including oleic acid and linoleic acid from 1090 to 1330 nm. Different pure fatty acids presented slightly-different THG χ(3) spectra. The measured peak values of THG third-order susceptibility χ(3) in both fatty acids were surprisingly found not to match completely with the resonant absorption wavelengths around 1190 to 1210 nm, suggesting possible wavelengths selection for enhanced THG imaging of lipids while avoiding laser light absorption. Along with the recent advancement in THG imaging, this new window between 1240 to 1290 nm may offer tremendous new opportunities for sensitive label-free lipid imaging in biological tissues.

Label-Free 3D Visualization of Cellular and Tissue Structures in Intact Muscle with Second and Third Harmonic Generation Microscopy

PubMed Central

Rehberg, Markus; Krombach, Fritz; Pohl, Ulrich; Dietzel, Steffen

2011-01-01

Second and Third Harmonic Generation (SHG and THG) microscopy is based on optical effects which are induced by specific inherent physical properties of a specimen. As a multi-photon laser scanning approach which is not based on fluorescence it combines the advantages of a label-free technique with restriction of signal generation to the focal plane, thus allowing high resolution 3D reconstruction of image volumes without out-of-focus background several hundred micrometers deep into the tissue. While in mammalian soft tissues SHG is mostly restricted to collagen fibers and striated muscle myosin, THG is induced at a large variety of structures, since it is generated at interfaces such as refraction index changes within the focal volume of the excitation laser. Besides, colorants such as hemoglobin can cause resonance enhancement, leading to intense THG signals. We applied SHG and THG microscopy to murine (Mus musculus) muscles, an established model system for physiological research, to investigate their potential for label-free tissue imaging. In addition to collagen fibers and muscle fiber substructure, THG allowed us to visualize blood vessel walls and erythrocytes as well as white blood cells adhering to vessel walls, residing in or moving through the extravascular tissue. Moreover peripheral nerve fibers could be clearly identified. Structure down to the nuclear chromatin distribution was visualized in 3D and with more detail than obtainable by bright field microscopy. To our knowledge, most of these objects have not been visualized previously by THG or any label-free 3D approach. THG allows label-free microscopy with inherent optical sectioning and therefore may offer similar improvements compared to bright field microscopy as does confocal laser scanning microscopy compared to conventional fluorescence microscopy. PMID:22140560

Molecular imaging of melanin distribution in vivo and quantitative differential diagnosis of human pigmented lesions using label-free harmonic generation biopsy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Sun, Chi-Kuang; Wei, Ming-Liang; Su, Yu-Hsiang; Weng, Wei-Hung; Liao, Yi-Hua

2017-02-01

Harmonic generation microscopy is a noninvasive repetitive imaging technique that provides real-time 3D microscopic images of human skin with a sub-femtoliter resolution and high penetration down to the reticular dermis. In this talk, we show that with a strong resonance effect, the third-harmonic-generation (THG) modality provides enhanced contrast on melanin and allows not only differential diagnosis of various pigmented skin lesions but also quantitative imaging for longterm tracking. This unique capability makes THG microscopy the only label-free technique capable of identifying the active melanocytes in human skin and to image their different dendriticity patterns. In this talk, we will review our recent efforts to in vivo image melanin distribution and quantitatively diagnose pigmented skin lesions using label-free harmonic generation biopsy. This talk will first cover the spectroscopic study on the melanin enhanced THG effect in human cells and the calibration strategy inside human skin for quantitative imaging. We will then review our recent clinical trials including: differential diagnosis capability study on pigmented skin tumors; as well as quantitative virtual biopsy study on pre- and post- treatment evaluation on melasma and solar lentigo. Our study indicates the unmatched capability of harmonic generation microscopy to perform virtual biopsy for noninvasive histopathological diagnosis of various pigmented skin tumors, as well as its unsurpassed capability to noninvasively reveal the pathological origin of different hyperpigmentary diseases on human face as well as to monitor the efficacy of laser depigmentation treatments. This work is sponsored by National Health Research Institutes.

Imaging of targeted lipid microbubbles to detect cancer cells using third harmonic generation microscopy

PubMed Central

Harpel, Kaitlin; Baker, Robert Dawson; Amirsolaimani, Babak; Mehravar, Soroush; Vagner, Josef; Matsunaga, Terry O.; Banerjee, Bhaskar; Kieu, Khanh

2016-01-01

The use of receptor-targeted lipid microbubbles imaged by ultrasound is an innovative method of detecting and localizing disease. However, since ultrasound requires a medium between the transducer and the object being imaged, it is impractical to apply to an exposed surface in a surgical setting where sterile fields need be maintained and ultrasound gel may cause the bubbles to collapse. Multiphoton microscopy (MPM) is an emerging tool for accurate, label-free imaging of tissues and cells with high resolution and contrast. We have recently determined a novel application of MPM to be used for detecting targeted microbubble adherence to the upregulated plectin-receptor on pancreatic tumor cells. Specifically, the third-harmonic generation response can be used to detect bound microbubbles to various cell types presenting MPM as an alternative and useful imaging method. This is an interesting technique that can potentially be translated as a diagnostic tool for the early detection of cancer and inflammatory disorders. PMID:27446711

Long-term imaging of mouse embryos using adaptive harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Thayil, Anisha; Watanabe, Tomoko; Jesacher, Alexander; Wilson, Tony; Srinivas, Shankar; Booth, Martin

2011-04-01

We present a detailed description of an adaptive harmonic generation (HG) microscope and culture techniques that permit long-term, three-dimensional imaging of mouse embryos. HG signal from both pre- and postimplantation stage (0.5-5.5 day-old) mouse embryos are fully characterized. The second HG images reveal central spindles during cytokinesis whereas third HG images show several features, such as lipid droplets, nucleoli, and plasma membranes. The embryos are found to develop normally during one-day-long discontinuous HG imaging, permitting the observation of several dynamic events, such as morula compaction and blastocyst formation.

Third harmonic generation in air ambient and laser ablated carbon plasma

DOE Office of Scientific and Technical Information (OSTI.GOV)

Singh, Ravi Pratap, E-mail: ravips@iitk.ac.in; Gupta, Shyam L.; Thareja, Raj K.

2015-12-15

We report the third harmonic generation of a nanosecond laser pulse (1.06 μm) in air ambient and in the presence of nanoparticles from laser ablated carbon plasma. Significant decrease in the threshold of third harmonic generation and multi-fold increment in the intensity of generated third harmonic is observed in presence of carbon plasma. The third harmonic in air is due to the quasi-resonant four photon process involving vibrationally excited states of molecular ion of nitrogen due to electron impact ionization and laser pulse. Following optical emission spectroscopic observations we conclude that the presence of C{sub 2} and CN in the ablatedmore » plume play a vital role in the observed third harmonic signals.« less

Three-dimensional image formation in fiber-optical second-harmonic-generation microscopy.

PubMed

Gu, Min; Fu, Ling

2006-02-06

Three-dimensional (3-D) image formation in fiber-optical second-harmonic-generation microscopy is revealed to be purely coherent and therefore can be described by a 3-D coherent transfer function (CTF) that exhibits the same spatial frequency passband as that of fiber-optical reflection-mode non-fluorescence microscopy. When the numerical aperture of the fiber is much larger than the angle of convergence of the illumination on the fiber aperture, the performance of fiber-optical second-harmonic-generation microscopy behaves as confocal second-harmonic-generation microscopy. The dependence of axial resolution on fiber coupling parameters shows an improvement of approximately 7%, compared with that in fiber-optical two-photon fluorescence microscopy.

Signal improvement in multiphoton microscopy by reflection with simple mirrors near the sample

NASA Astrophysics Data System (ADS)

Rehberg, Markus; Krombach, Fritz; Pohl, Ulrich; Dietzel, Steffen

2010-03-01

In conventional fluorescence or confocal microscopy, emitted light is generated not only in the focal plane but also above and below. The situation is different in multiphoton-induced fluorescence and multiphoton-induced higher harmonic generation. Here, restriction of signal generation to a single focal point permits that all emitted photons can contribute to image formation if collected, regardless of their path through the specimen. Often, the intensity of the emitted light is rather low in biological specimens. We present a method to significantly increase the fraction of photons collected by an epi (backward) detector by placing a simple mirror, an aluminum-coated coverslip, directly under the sample. Samples investigated include fluorescent test slides, collagen gels, and thin-layered, intact mouse skeletal muscles. Quantitative analysis revealed an intensity increase of second- and third-harmonic generated signal in skeletal muscle of nine- and sevenfold respectively, and of fluorescent signal in test slides of up to twofold. Our approach thus allows significant signal improvement also for situations were a forward detection is impossible, e.g., due to the anatomy of animals in intravital microscopy.

Differentiating intratumoral melanocytes from Langerhans cells in nonmelanocytic pigmented skin tumors in vivo by label-free third-harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Weng, Wei-Hung; Liao, Yi-Hua; Tsai, Ming-Rung; Wei, Ming-Liang; Huang, Hsin-Yi; Sun, Chi-Kuang

2016-07-01

Morphology and distribution of melanocytes are critical imaging information for the diagnosis of melanocytic lesions. However, how to image intratumoral melanocytes noninvasively in pigmented skin tumors is seldom investigated. Third-harmonic generation (THG) is shown to be enhanced by melanin, whereas high accuracy has been demonstrated using THG microscopy for in vivo differential diagnosis of nonmelanocytic pigmented skin tumors. It is thus desirable to investigate if label-free THG microscopy was capable to in vivo identify intratumoral melanocytes. In this study, histopathological correlations of label-free THG images with the immunohistochemical images stained with human melanoma black (HMB)-45 and cluster of differentiation 1a (CD1a) were made. The correlation results indicated that the intratumoral THG-bright dendritic-cell-like signals were endogenously derived from melanocytes rather than Langerhans cells (LCs). The consistency between THG-bright dendritic-cell-like signals and HMB-45 melanocyte staining showed a kappa coefficient of 0.807, 84.6% sensitivity, and 95% specificity. In contrast, a kappa coefficient of -0.37, 21.7% sensitivity, and 30% specificity were noted between the THG-bright dendritic-cell-like signals and CD1a staining for LCs. Our study indicates the capability of noninvasive label-free THG microscopy to differentiate intratumoral melanocytes from LCs, which is not feasible in previous in vivo label-free clinical-imaging modalities.

Simultaneous stimulated Raman scattering and higher harmonic generation imaging for liver disease diagnosis without labeling

NASA Astrophysics Data System (ADS)

Lin, Jian; Wang, Zi; Zheng, Wei; Huang, Zhiwei

2014-02-01

Nonlinear optical microscopy (e.g., higher harmonic (second-/third- harmonic) generation (HHG), simulated Raman scattering (SRS)) has high diagnostic sensitivity and chemical specificity, making it a promising tool for label-free tissue and cell imaging. In this work, we report a development of a simultaneous SRS and HHG imaging technique for characterization of liver disease in a bile-duct-ligation rat-modal. HHG visualizes collagens formation and reveals the cell morphologic changes associated with liver fibrosis; whereas SRS identifies the distributions of hepatic fat cells formed in steatosis liver tissue. This work shows that the co-registration of SRS and HHG images can be an effective means for label-free diagnosis and characterization of liver steatosis/fibrosis at the cellular and molecular levels.

The analysis of harmonic generation coefficients in the ablative Rayleigh-Taylor instability

NASA Astrophysics Data System (ADS)

Lu, Yan; Fan, Zhengfeng; Lu, Xinpei; Ye, Wenhua; Zou, Changlin; Zhang, Ziyun; Zhang, Wen

2017-10-01

In this research, we use the numerical simulation method to investigate the generation coefficients of the first three harmonics and the zeroth harmonic in the Ablative Rayleigh-Taylor Instability. It is shown that the interface shifts to the low temperature side during the ablation process. In consideration of the third-order perturbation theory, the first three harmonic amplitudes of the weakly nonlinear regime are calculated and then the harmonic generation coefficients are obtained by curve fitting. The simulation results show that the harmonic generation coefficients changed with time and wavelength. Using the higher-order perturbation theory, we find that more and more harmonics are generated in the later weakly nonlinear stage, which is caused by the negative feedback of the later higher harmonics. Furthermore, extending the third-order theory to the fifth-order theory, we find that the second and the third harmonics coefficients linearly depend on the wavelength, while the feedback coefficients are almost constant. Further analysis also shows that when the fifth-order theory is considered, the normalized effective amplitudes of second and third harmonics can reach about 25%-40%, which are only 15%-25% in the frame of the previous third-order theory. Therefore, the third order perturbation theory is needed to be modified by the higher-order theory when ηL reaches about 20% of the perturbation wavelength.

Wiggler magnetic field assisted third harmonic generation in expanding clusters

NASA Astrophysics Data System (ADS)

Vij, Shivani

2018-04-01

A simple theoretical model is constructed to study the wiggler magnetic field assisted third harmonic generation of intense short pulse laser in a cluster in its expanding phase. The ponderomotive force of laser causes density perturbations in cluster electron density which couples with wiggler magnetic field to produce a nonlinear current that generates transverse third harmonic. An intense short pulse laser propagating through a gas embedded with atomic clusters, converts it into hot plasma balls via tunnel ionization. Initially, the electron plasma frequency inside the clusters ω pe > \\sqrt{3}{ω }1 (with ω 1 being the frequency of the laser). As the cluster expands under Coulomb force and hydrodynamic pressure, ω pe decreases to \\sqrt{3}{ω }1. At this time, there is resonant enhancement in the efficiency of the third harmonic generation. The efficiency of third harmonic generation is enhanced due to cluster plasmon resonance and by phase matching due to wiggler magnetic field. The effect of cluster size on the expansion rate is studied to observe that the clusters of different radii would expand differently. The impact of laser intensity and wiggler magnetic field on the efficiency of third harmonic generation is also explored.

Quantitative changes in human epithelial cancers and osteogenesis imperfecta disease detected using nonlinear multicontrast microscopy

NASA Astrophysics Data System (ADS)

Adur, Javier; Pelegati, Vitor B.; de Thomaz, Andre A.; D'Souza-Li, Lilia; Assunção, Maria do Carmo; Bottcher-Luiz, Fátima; Andrade, Liliana A. L. A.; Cesar, Carlos L.

2012-08-01

We show that combined multimodal nonlinear optical (NLO) microscopies, including two-photon excitation fluorescence, second-harmonic generation (SHG), third harmonic generation, and fluorescence lifetime imaging microscopy (FLIM) can be used to detect morphological and metabolic changes associated with stroma and epithelial transformation during the progression of cancer and osteogenesis imperfecta (OI) disease. NLO microscopes provide complementary information about tissue microstructure, showing distinctive patterns for different types of human breast cancer, mucinous ovarian tumors, and skin dermis of patients with OI. Using a set of scoring methods (anisotropy, correlation, uniformity, entropy, and lifetime components), we found significant differences in the content, distribution and organization of collagen fibrils in the stroma of breast and ovary as well as in the dermis of skin. We suggest that our results provide a framework for using NLO techniques as a clinical diagnostic tool for human cancer and OI. We further suggest that the SHG and FLIM metrics described could be applied to other connective or epithelial tissue disorders that are characterized by abnormal cells proliferation and collagen assembly.

Investigation of phase matching for third-harmonic generation in silicon slow light photonic crystal waveguides using Fourier optics.

PubMed

Monat, Christelle; Grillet, Christian; Corcoran, Bill; Moss, David J; Eggleton, Benjamin J; White, Thomas P; Krauss, Thomas F

2010-03-29

Using Fourier optics, we retrieve the wavevector dependence of the third-harmonic (green) light generated in a slow light silicon photonic crystal waveguide. We show that quasi-phase matching between the third-harmonic signal and the fundamental mode is provided in this geometry by coupling to the continuum of radiation modes above the light line. This process sustains third-harmonic generation with a relatively high efficiency and a substantial bandwidth limited only by the slow light window of the fundamental mode. The results give us insights into the physics of this nonlinear process in the presence of strong absorption and dispersion at visible wavelengths where bandstructure calculations are problematic. Since the characteristics (e.g. angular pattern) of the third-harmonic light primarily depend on the fundamental mode dispersion, they could be readily engineered.

Contribution of the magnetic resonance to the third harmonic generation from a fishnet metamaterial

NASA Astrophysics Data System (ADS)

Reinhold, J.; Shcherbakov, M. R.; Chipouline, A.; Panov, V. I.; Helgert, C.; Paul, T.; Rockstuhl, C.; Lederer, F.; Kley, E.-B.; Tünnermann, A.; Fedyanin, A. A.; Pertsch, T.

2012-09-01

We investigate experimentally and theoretically the third harmonic generated by a double-layer fishnet metamaterial. To unambiguously disclose most notably the influence of the magnetic resonance, the generated third harmonic was measured as a function of the angle of incidence. It is shown experimentally and numerically that when the magnetic resonance is excited by a pump beam, the angular dependence of the third harmonic signal has a local maximum at an incidence angle of θ≃20∘. This maximum is shown to be a fingerprint of the antisymmetric distribution of currents in the gold layers. An analytical model based on the nonlinear dynamics of the electrons inside the gold shows excellent agreement with experimental and numerical results. This clearly indicates the difference in the third harmonic angular pattern at electric and magnetic resonances of the metamaterial.

Raman parametric excitation effect upon the third harmonic generation by a metallic nanoparticle lattice

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sepehri Javan, N., E-mail: sepehri-javan@uma.ac.ir

2015-08-21

This work is a theoretical study on third harmonic generation in the nonlinear propagation of an intense laser pulse through a periodic three-dimensional lattice of nanoparticles. Using a perturbative method, the nonlinear equations that describe the laser–nanoparticle interaction in the weakly relativistic regime are derived. Additionally, the nonlinear dispersion relation and the amplitude of the third harmonic are obtained. Finally, the effects of the nanoparticle radius and separation length, the distribution of the nanoparticle electron density, and the laser frequency upon the third harmonic efficiency are investigated. In addition to the expected resonance that occurs when the third harmonic resonatesmore » with the plasmon wave, another resonance appears when the nonlinear interaction of the fundamental mode with the third harmonic excites a longitudinal collective plasmon wave via the parametric Raman mechanism.« less

Quantitative comparison of 3D third harmonic generation and fluorescence microscopy images.

PubMed

Zhang, Zhiqing; Kuzmin, Nikolay V; Groot, Marie Louise; de Munck, Jan C

2018-01-01

Third harmonic generation (THG) microscopy is a label-free imaging technique that shows great potential for rapid pathology of brain tissue during brain tumor surgery. However, the interpretation of THG brain images should be quantitatively linked to images of more standard imaging techniques, which so far has been done qualitatively only. We establish here such a quantitative link between THG images of mouse brain tissue and all-nuclei-highlighted fluorescence images, acquired simultaneously from the same tissue area. For quantitative comparison of a substantial pair of images, we present here a segmentation workflow that is applicable for both THG and fluorescence images, with a precision of 91.3 % and 95.8 % achieved respectively. We find that the correspondence between the main features of the two imaging modalities amounts to 88.9 %, providing quantitative evidence of the interpretation of dark holes as brain cells. Moreover, 80 % bright objects in THG images overlap with nuclei highlighted in the fluorescence images, and they are 2 times smaller than the dark holes, showing that cells of different morphologies can be recognized in THG images. We expect that the described quantitative comparison is applicable to other types of brain tissue and with more specific staining experiments for cell type identification. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

In-vivo third-harmonic generation microscopy at 1550nm three-dimensional long-term time-lapse studies in living C. elegans embryos

NASA Astrophysics Data System (ADS)

Aviles-Espinosa, Rodrigo; Santos, Susana I. C. O.; Brodschelm, Andreas; Kaenders, Wilhelm G.; Alonso-Ortega, Cesar; Artigas, David; Loza-Alvarez, Pablo

2011-03-01

In-vivo microscopic long term time-lapse studies require controlled imaging conditions to preserve sample viability. Therefore it is crucial to meet specific exposure conditions as these may limit the applicability of established techniques. In this work we demonstrate the use of third harmonic generation (THG) microscopy for long term time-lapse three-dimensional studies (4D) in living Caenorhabditis elegans embryos employing a 1550 nm femtosecond fiber laser. We take advantage of the fact that THG only requires the existence of interfaces to generate signal or a change in the refractive index or in the χ3 nonlinear coefficient, therefore no markers are required. In addition, by using this wavelength the emitted THG signal is generated at visible wavelengths (516 nm) enabling the use of standard collection optics and detectors operating near their maximum efficiency. This enables the reduction of the incident light intensity at the sample plane allowing to image the sample for several hours. THG signal is obtained through all embryo development stages, providing different tissue/structure information. By means of control samples, we demonstrate that the expected water absorption at this wavelength does not severely compromise sample viability. Certainly, this technique reduces the complexity of sample preparation (i.e. genetic modification) required by established linear and nonlinear fluorescence based techniques. We demonstrate the non-invasiveness, reduced specimen interference, and strong potential of this particular wavelength to be used to perform long-term 4D recordings.

Intravital third harmonic generation microscopy of collective melanoma cell invasion

PubMed Central

Weigelin, Bettina; Bakker, Gert-Jan; Friedl, Peter

2012-01-01

Cancer cell invasion is an adaptive process based on cell-intrinsic properties to migrate individually or collectively, and their adaptation to encountered tissue structure acting as barrier or providing guidance. Whereas molecular and physical mechanisms of cancer invasion are well-studied in 3D in vitro models, their topographic relevance, classification and validation toward interstitial tissue organization in vivo remain incomplete. Using combined intravital third and second harmonic generation (THG, SHG), and three-channel fluorescence microscopy in live tumors, we here map B16F10 melanoma invasion into the dermis with up to 600 µm penetration depth and reconstruct both invasion mode and tissue tracks to establish invasion routes and outcome. B16F10 cells preferentially develop adaptive invasion patterns along preformed tracks of complex, multi-interface topography, combining single-cell and collective migration modes, without immediate anatomic tissue remodeling or destruction. The data suggest that the dimensionality (1D, 2D, 3D) of tissue interfaces determines the microanatomy exploited by invading tumor cells, emphasizing non-destructive migration along microchannels coupled to contact guidance as key invasion mechanisms. THG imaging further detected the presence and interstitial dynamics of tumor-associated microparticles with submicron resolution, revealing tumor-imposed conditioning of the microenvironment. These topographic findings establish combined THG, SHG and fluorescence microscopy in intravital tumor biology and provide a template for rational in vitro model development and context-dependent molecular classification of invasion modes and routes. PMID:29607252

Optical Biomarkers of Serous and Mucinous Human Ovarian Tumor Assessed with Nonlinear Optics Microscopies

PubMed Central

Adur, Javier; Pelegati, Vitor B.; de Thomaz, Andre A.; Baratti, Mariana O.; Almeida, Diogo B.; Andrade, L. A. L. A.; Bottcher-Luiz, Fátima; Carvalho, Hernandes F.; Cesar, Carlos L.

2012-01-01

Background Nonlinear optical (NLO) microscopy techniques have potential to improve the early detection of epithelial ovarian cancer. In this study we showed that multimodal NLO microscopies, including two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), third-harmonic generation (THG) and fluorescence lifetime imaging microscopy (FLIM) can detect morphological and metabolic changes associated with ovarian cancer progression. Methodology/Principal Findings We obtained strong TPEF + SHG + THG signals from fixed samples stained with Hematoxylin & Eosin (H&E) and robust FLIM signal from fixed unstained samples. Particularly, we imaged 34 ovarian biopsies from different patients (median age, 49 years) including 5 normal ovarian tissue, 18 serous tumors and 11 mucinous tumors with the multimodal NLO platform developed in our laboratory. We have been able to distinguish adenomas, borderline, and adenocarcinomas specimens. Using a complete set of scoring methods we found significant differences in the content, distribution and organization of collagen fibrils in the stroma as well as in the morphology and fluorescence lifetime from epithelial ovarian cells. Conclusions/Significance NLO microscopes provide complementary information about tissue microstructure, showing distinctive patterns for serous and mucinous ovarian tumors. The results provide a basis to interpret future NLO images of ovarian tissue and lay the foundation for future in vivo optical evaluation of premature ovarian lesions. PMID:23056557

Simultaneously phase-matched second- and third-harmonic generation from 1.55 microm radiation in annealed proton-exchanged periodically poled lithium niobate waveguides.

PubMed

Marangoni, M; Lobino, M; Ramponi, R

2006-09-15

Third-harmonic generation (THG) in the cw regime from C-band radiation was achieved in annealed proton-exchanged periodically poled lithium niobate (PPLN) waveguides. By suitable design of fabrication parameters and operating conditions, quasi-phase-matching (QPM) is obtained simultaneously for the second-harmonic generation process (omega-->2omega, first-order QPM) and for the sum-frequency-generation process (omega+2omega-->3omega, third-order QPM), which provides the third harmonic of the pump field. The high overlap between the field profiles of the interacting modes--TM00 at omega and TM10 at 2omega and 3omega--results in what is believed to be the highest ever reported normalized conversion efficiency for THG from telecommunication wavelengths, equal to 0.72%W(-2) cm(-4).

Ince-gauss based multiple intermodal phase-matched third-harmonic generations in a step-index silica optical fiber

NASA Astrophysics Data System (ADS)

Borne, Adrien; Katsura, Tomotaka; Félix, Corinne; Doppagne, Benjamin; Segonds, Patricia; Bencheikh, Kamel; Levenson, Juan Ariel; Boulanger, Benoit

2016-01-01

Several third-harmonic generation processes were performed in a single step-index germanium-doped silica optical fiber under intermodal phase-matching conditions. The nanosecond fundamental beam range between 1400 and 1600 nm. The transverse distributions of the energy were successfully modeled in the form of Ince-Gauss modes, pointing out some ellipticity of fiber core. From these experiments and theoretical calculations, we discuss the implementation of frequency degenerated triple photon generation that shares the same phase-matching condition as third-harmonic generation, which is its reverse process.

Bernstein wave aided laser third harmonic generation in a plasma

NASA Astrophysics Data System (ADS)

Tyagi, Yachna; Tripathi, Deepak; Kumar, Ashok

2016-09-01

The process of Bernstein wave aided resonant third harmonic generation of laser in a magnetized plasma is investigated. The extra-ordinary mode (X-mode) laser of frequency ω 0 and wave number k → 0 , travelling across the magnetic field in a plasma, exerts a second harmonic ponderomotive force on the electrons imparting them an oscillatory velocity v → 2 ω0 , 2 k → 0 . This velocity beats with the density perturbation due to the Bernstein wave to produce a density perturbation at cyclotron frequency shifted second harmonic. The density perturbation couples with the oscillatory velocity v → ω0 , k → 0 of X-mode of the laser to produce the cyclotron frequency shifted third harmonic current density leading to harmonic radiation. The phase matching condition for the up shifted frequency is satisfied when the Bernstein wave is nearly counter-propagating to the laser. As the transverse wave number of the Bernstein wave is large, it is effective in the phase matched third harmonic generation, when the laser frequency is not too far from the upper hybrid frequency.

Effects of electromagnetic fields on the nonlinear optical properties of asymmetric double quantum well under intense laser field

NASA Astrophysics Data System (ADS)

Yesilgul, U.; Sari, H.; Ungan, F.; Martínez-Orozco, J. C.; Restrepo, R. L.; Mora-Ramos, M. E.; Duque, C. A.; Sökmen, I.

2017-03-01

In this study, the effects of electric and magnetic fields on the optical rectification and second and third harmonic generation in asymmetric double quantum well under the intense non-resonant laser field is theoretically investigated. We calculate the optical rectification and second and third harmonic generation within the compact density-matrix approach. The theoretical findings show that the influence of electric, magnetic, and intense laser fields leads to significant changes in the coefficients of nonlinear optical rectification, second and third harmonic generation.

Taking a deep look: modern microscopy technologies to optimize the design and functionality of biocompatible scaffolds for tissue engineering in regenerative medicine

PubMed Central

Vielreicher, M.; Schürmann, S.; Detsch, R.; Schmidt, M. A.; Buttgereit, A.; Boccaccini, A.; Friedrich, O.

2013-01-01

This review focuses on modern nonlinear optical microscopy (NLOM) methods that are increasingly being used in the field of tissue engineering (TE) to image tissue non-invasively and without labelling in depths unreached by conventional microscopy techniques. With NLOM techniques, biomaterial matrices, cultured cells and their produced extracellular matrix may be visualized with high resolution. After introducing classical imaging methodologies such as µCT, MRI, optical coherence tomography, electron microscopy and conventional microscopy two-photon fluorescence (2-PF) and second harmonic generation (SHG) imaging are described in detail (principle, power, limitations) together with their most widely used TE applications. Besides our own cell encapsulation, cell printing and collagen scaffolding systems and their NLOM imaging the most current research articles will be reviewed. These cover imaging of autofluorescence and fluorescence-labelled tissue and biomaterial structures, SHG-based quantitative morphometry of collagen I and other proteins, imaging of vascularization and online monitoring techniques in TE. Finally, some insight is given into state-of-the-art three-photon-based imaging methods (e.g. coherent anti-Stokes Raman scattering, third harmonic generation). This review provides an overview of the powerful and constantly evolving field of multiphoton microscopy, which is a powerful and indispensable tool for the development of artificial tissues in regenerative medicine and which is likely to gain importance also as a means for general diagnostic medical imaging. PMID:23864499

Optical harmonic generator

DOEpatents

Summers, M.A.; Eimerl, D.; Boyd, R.D.

1982-06-10

A pair of uniaxial birefringent crystal elements are fixed together to form a serially arranged, integral assembly which, alternatively, provides either a linearly or elliptically polarized second-harmonic output wave or a linearly polarized third-harmonic output wave. The extraordinary or e directions of the crystal elements are oriented in the integral assembly to be in quadrature (90/sup 0/). For a second-harmonic generation in the Type-II-Type-II angle tuned case, the input fundamental wave has equal amplitude o and e components. For a third-harmonic generation, the input fundamental wave has o and e components whose amplitudes are in a ratio of 2:1 (o:e reference first crystal). In the typical case of a linearly polarized input fundamental wave this can be accomplished by simply rotating the crystal assembly about the input beam direction by 10/sup 0/. For both second and third harmonic generation input precise phase-matching is achieved by tilting the crystal assembly about its two sensitive axeses (o).

Optical harmonic generator

DOEpatents

Summers, Mark A.; Eimerl, David; Boyd, Robert D.

1985-01-01

A pair of uniaxial birefringent crystal elements are fixed together to form a serially arranged, integral assembly which, alternatively, provides either a linearly or elliptically polarized second-harmonic output wave or a linearly polarized third-harmonic output wave. The "extraordinary" or "e" directions of the crystal elements are oriented in the integral assembly to be in quadrature (90.degree.). For a second-harmonic generation in the Type-II-Type-II angle tuned case, the input fundamental wave has equal amplitude "o" and "e" components. For a third-harmonic generation, the input fundamental wave has "o" and "e" components whose amplitudes are in a ratio of 2:1 ("o":"e" reference first crystal). In the typical case of a linearly polarized input fundamental wave this can be accomplished by simply rotating the crystal assembly about the input beam direction by 10.degree.. For both second and third harmonic generation input precise phase-matching is achieved by tilting the crystal assembly about its two sensitive axes ("o").

Studying the effect of photodynamic therapy (PDT) to enhance healing of femur fractures using polarimetric second-harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Golaraei, Ahmad; Raja, Vaishnavi; Akens, Margarete K.; Wilson, Brian C.; Barzda, Virginijus

2017-07-01

Linear polarization-in, polarization-out second-harmonic generation microscopy was used to study the effect of Photodynamic therapy treatment on enhancing the healing of femur fracture by investigating the ultrastructure of collagen as a major component of bone matrix.

Inhibition of linear absorption in opaque materials using phase-locked harmonic generation.

PubMed

Centini, Marco; Roppo, Vito; Fazio, Eugenio; Pettazzi, Federico; Sibilia, Concita; Haus, Joseph W; Foreman, John V; Akozbek, Neset; Bloemer, Mark J; Scalora, Michael

2008-09-12

We theoretically predict and experimentally demonstrate inhibition of linear absorption for phase and group velocity mismatched second- and third-harmonic generation in strongly absorbing materials, GaAs, in particular, at frequencies above the absorption edge. A 100-fs pump pulse tuned to 1300 nm generates 650 and 435 nm second- and third-harmonic pulses that propagate across a 450-microm-thick GaAs substrate without being absorbed. We attribute this to a phase-locking mechanism that causes the pump to trap the harmonics and to impress on them its dispersive properties.

Ultrafast third-harmonic generation from textured aluminum nitride-sapphire interfaces

NASA Astrophysics Data System (ADS)

Stoker, D. S.; Baek, J.; Wang, W.; Kovar, D.; Becker, M. F.; Keto, J. W.

2006-05-01

We measured and modeled third-harmonic generation (THG) from an AlN thin film on sapphire using a time-domain approach appropriate for ultrafast lasers. Second-harmonic measurements indicated that polycrystalline AlN contains long-range crystal texture. An interface model for third-harmonic generation enabled an analytical representation of scanning THG ( z -scan) experiments. Using it and accounting for Fresnel reflections, we measured the AlN -sapphire susceptibility ratio and estimated the susceptibility for aluminum nitride, χxxxx(3)(3ω;ω,ω,ω)=1.52±0.25×10-13esu . The third-harmonic (TH) spectrum strongly depended on the laser focus position and sample thickness. The amplitude and phase of the frequency-domain interference were fit to the Fourier transform of the calculated time-domain field to improve the accuracy of several experimental parameters. We verified that the model works well for explaining TH signal amplitudes and spectral phase. Some anomalous features in the TH spectrum were observed, which we attributed to nonparaxial effects.

Phase-matched second- and third-harmonic generation in plasmas with density ripple

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dahiya, Deepak; Sajal, Vivek; Sharma, A. K.

The generation of second and third harmonics by the interaction of an ultrashort laser pulse with underdense plasma having a density ripple is studied at intensities I{lambda}{sup 2}=10{sup 16}-10{sup 19} W cm{sup -2} {mu}m{sup 2} using fully relativistic two-dimensional particle-in-cell simulations with high spectral resolution. A theoretical model is developed for second- and third-harmonic conversion efficiencies. When the laser is plane polarized in the simulation plane even and odd harmonics are excited in the same polarization as the laser polarization. The highest efficiency of generation of a specific harmonic occurs when the ripple wave vector value k{sub q} satisfies phase-matchingmore » conditions. The efficiency of phase-matched harmonic generation is an order of magnitude higher than the one without phase matching. The efficiency increases rapidly in weak and moderate relativistic regime and tends to saturate in strong relativistic regime. At moderately relativistic intensities and low plasma densities, the simulation and recent experimental results are fairly reproduced by an analytical theory.« less

Characterizing Fibrosis in Mouse Kidney using Label Free Fluorescence Lifetime and Second Harmonic Generation Imaging Microscopy in Unilateral Ureteral Obstruction Model

PubMed Central

Ranjit, Suman; Dobrinskikh, Evgenia; Montford, John; Dvornikov, Alexander; Lehman, Allison; Orlicky, David J.; Nemenoff, Raphael; Gratton, Enrico; Levi, Moshe; Furgeson, Seth

2017-01-01

All forms of progressive renal diseases develop a final pathway of tubulointerstitial fibrosis and glomerulosclerosis. Renal fibrosis is usually quantified using histological staining, a process that is time-consuming and pathologist dependent. The work described here shows the development of a fast and operator-independent method to measure fibrosis. To study renal fibrosis, the unilateral ureteral obstruction (UUO) model was chosen. Mice develop a time-dependent increase in obstructed kidneys; contralateral kidneys are used as controls. After UUO, kidneys were analyzed at three time points: 7 days, 14 days, and 21 days. Fibrosis was investigated using FLIM (Fluorescence Lifetime Imaging) and SHG (Second Harmonic Generation) in the deep tissue imaging microscope called DIVER (Deep Imaging via Enhanced photon Recovery). This microscope was developed for deep tissue and SHG and THG (Third Harmonic Generation) imaging and has extraordinary sensitivity towards harmonic generation. SHG data suggests the presence of more fibrillar collagen in the diseased kidneys. The combinations of short wavelength FLIM and SHG analysis results in a robust analysis procedure independent of observer interpretation and let us create a criterion to quantify the extent of fibrosis directly from the image. The progression of fibrosis in UUO model has been studied using this new FLIM-SHG technique and it shows remarkable improvement in quantification of fibrosis compared to standard histological techniques. PMID:27555119

Hybrid microscopy of human carotid atheroma by means of optical-resolution optoacoustic and non-linear optical microscopy

NASA Astrophysics Data System (ADS)

Seeger, Markus; Karlas, Angelos; Soliman, Dominik; Pelisek, Jaroslav; Ntziachristos, Vasilis

2017-03-01

Carotid atheromatosis is causally related to stroke, a leading cause of disability and death. We present the analysis of a human carotid atheroma using a novel hybrid microscopy system that combines optical-resolution optoacoustic (photoacoustic) microscopy and several non-linear optical microscopy modalities (second and third harmonic generation, as well as, two-photon excitation fluorescence) to achieve a multimodal examination of the extracted tissue within the same imaging framework. Our system enables the label-free investigation of atheromatous human carotid tissue with a resolution of about 1 μm and allows for the congruent interrogation of plaque morphology and clinically relevant constituents such as red blood cells, collagen, and elastin. Our data reveal mutual interactions between blood embeddings and connective tissue within the atheroma, offering comprehensive insights into its stage of evolution and severity, and potentially facilitating the further development of diagnostic tools, as well as treatment strategies.

Simultaneous quadruple modal nonlinear optical imaging for gastric diseases diagnosis and characterization

NASA Astrophysics Data System (ADS)

Wang, Zi; Zheng, Wei; Lin, Jian; Huang, Zhiwei

2015-03-01

We report the development of a unique simultaneous quadruple-modal nonlinear optical microscopy (i.e., stimulated Raman scattering (SRS), second-harmonic generation (SHG), two-photon excitation fluorescence (TPEF), and third-harmonic generation (THG)) platform for characterization of the gastric diseases (i.e., gastritis, intestinal metaplasia (IM), intestinal type adenocarcinoma). SRS highlights the goblet cells found in IM. SHG images the distribution of collagen in lamina propria. Collagen is found to aggregate for intestinal type adenocarcinoma. TPEF reveals the cell morphology and can reflect the damage inside glands caused by the diseases. THG visualizes the nuclei with high spatial resolution, which facilitates the identification of neutrophils that are usually used as a feature of inflammation. This work shows that the co-registration of quadruple-modal images can be an effective means for diagnosis and characterization of gastric diseases at the cellular and molecular levels.

Module for multiphoton high-resolution hyperspectral imaging and spectroscopy

NASA Astrophysics Data System (ADS)

Zeytunyan, Aram; Baldacchini, Tommaso; Zadoyan, Ruben

2018-02-01

We developed a module for dual-output, dual-wavelength lasers that facilitates multiphoton imaging and spectroscopy experiments and enables hyperspectral imaging with spectral resolution up to 5 cm-1. High spectral resolution is achieved by employing spectral focusing. Specifically, two sets of grating pairs are used to control the chirps in each laser beam. In contrast with the approach that uses fixed-length glass rods, grating pairs allow matching the spectral resolution and the linewidths of the Raman lines of interest. To demonstrate the performance of the module, we report the results of spectral focusing CARS and SRS microscopy experiments for various test samples and Raman shifts. The developed module can be used for a variety of multimodal imaging and spectroscopy applications, such as single- and multi-color two-photon fluorescence, second harmonic generation, third harmonic generation, pump-probe, transient absorption, and others.

Dispersion tuning in sub-micron tapers for third-harmonic and photon triplet generation.

PubMed

Hammer, Jonas; Cavanna, Andrea; Pennetta, Riccardo; Chekhova, Maria V; Russell, Philip St J; Joly, Nicolas Y

2018-05-15

Precise control of the dispersion landscape is of crucial importance if optical fibers are to be successfully used for the generation of three-photon states of light-the inverse of third-harmonic generation (THG). Here we report gas-tuning of intermodal phase-matched THG in sub-micron-diameter tapered optical fiber. By adjusting the pressure of the surrounding argon gas up to 50 bars, intermodally phase-matched third-harmonic light can be generated for pump wavelengths within a 15 nm range around 1.38 μm. We also measure the infrared fluorescence generated in the fiber when pumped in the visible and estimate that the accidental coincidence rate in this signal is lower than the predicted detection rate of photon triplets.

Polarization anisotropy in fiber-optic second harmonic generation microscopy.

PubMed

Fu, Ling; Gu, Min

2008-03-31

We report the investigation and implementation of a compact second harmonic generation microscope that uses a single-mode fiber coupler and a double-clad photonic crystal fiber. Second harmonic polarization anisotropy through the fiber-optic microscope systems is quantitatively measured with KTP microcrystals, fish scale and rat tail tendon. It is demonstrated that the polarized second harmonic signals can be excited and collected through the single-mode fiber coupler to analyze the molecular orientations of structural proteins. It has been discovered that a double-clad photonic crystal fiber can preserve the linear polarization in the core, although a depolarization effect is observed in the inner cladding region. The feasibility of polarization anisotropy measurements in fiber-optic second harmonic generation microscopy will benefit the in vivo study of collagen-related diseases with a compact imaging probe.

Second and Third Harmonic Generation in Metal-Based Nanostructures

DTIC Science & Technology

2010-01-01

Prudenzano, D. de Ceglia, N. Akozbek, M.J. Bloemer, P. Ashley, and M. Scalora , "Enhanced transmission and second harmonic generation from...Fazio, C. Sibilia, M. J. Bloemer, and M. Scalora , "Second-harmonic generation from metallodielectric multilayer photonic-band-gap structures", Phys...harmonic generation", Phys. Rev. B 38, 7985 (1988). [50] M. A. Vincenti, D. de Ceglia, M. Buncick, N. Akozbek, M. J. Bloemer, and M. Scalora

All-fiber femtosecond laser providing 9 nJ, 50 MHz pulses at 1650 nm for three-photon microscopy

NASA Astrophysics Data System (ADS)

Cadroas, P.; Abdeladim, L.; Kotov, L.; Likhachev, M.; Lipatov, D.; Gaponov, D.; Hideur, A.; Tang, M.; Livet, J.; Supatto, W.; Beaurepaire, E.; Février, S.

2017-06-01

The spectral window lying between 1.6 and 1.7 μm is interesting for in-depth multiphoton microscopy of intact tissues due to reduced scattering and absorption in this wavelength range. However, wide adoption of this excitation range will rely on the availability of robust and cost-effective high peak power pulsed lasers operating at these wavelengths. In this communication, we report on a monolithically integrated high repetition rate (50 MHz) all-fiber femtosecond laser based on a soliton self-frequency shift providing 9 nJ, 75 fs pulses at 1650 nm. We illustrate its potential for biological microscopy by recording three-photon-excited fluorescence and third-harmonic generation images of mouse nervous tissue and developing Drosophila embryos labeled with a red fluorescent protein.

Measurement of absorption spectrum of deuterium oxide (D{sub 2}O) and its application to signal enhancement in multiphoton microscopy at the 1700-nm window

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Yuxin; Wen, Wenhui; Wang, Kai

2016-01-11

1700-nm window has been demonstrated to be a promising excitation window for deep-tissue multiphoton microscopy (MPM). Long working-distance water immersion objective lenses are typically used for deep-tissue imaging. However, absorption due to immersion water at 1700 nm is still high and leads to dramatic decrease in signals. In this paper, we demonstrate measurement of absorption spectrum of deuterium oxide (D{sub 2}O) from 1200 nm to 2600 nm, covering the three low water-absorption windows potentially applicable for deep-tissue imaging (1300 nm, 1700 nm, and 2200 nm). We apply this measured result to signal enhancement in MPM at the 1700-nm window. Compared with water immersion, D{sub 2}O immersionmore » enhances signal levels in second-harmonic generation imaging, 3-photon fluorescence imaging, and third-harmonic generation imaging by 8.1, 24.8, and 24.7 times with 1662-nm excitation, in good agreement with theoretical calculation based on our absorption measurement. This suggests D{sub 2}O a promising immersion medium for deep-tissue imaging.« less

A Compact Ti:Sapphire Laser With its Third Harmonic Generation (THG) for an Airborne Ozone Differential Absorption Lidar (DIAL) Transmitter

NASA Technical Reports Server (NTRS)

Chen, Songsheng; Storm, Mark E.; Marsh, Waverly D.; Petway, Larry B.; Edwards, William C.; Barnes, James C.

2000-01-01

A compact and high-pulse-energy Ti:Sapphire laser with its Third Harmonic Generation (THG) has been developed for an airborne ozone differential absorption lidar (DIAL) to study the distributions and concentrations of the ozone throughout the troposphere. The Ti:Sapphire laser, pumped by a frequency-doubled Nd:YAG laser and seeded by a single mode diode laser, is operated either at 867 nm or at 900 nm with a pulse repetition frequency of 20 Hz. High energy laser pulses (more than 110 mJ/pulse) at 867 nm or 900 nm with a desired beam quality have been achieved and utilized to generate its third harmonic at 289nm or 300nm, which are on-line and off-line wavelengths of an airborne ozone DIAL. After being experimentally compared with Beta-Barium Borate (beta - BaB2O4 or BBO) nonlinear crystals, two Lithium Triborate (LBO) crystals (5 x 5 x 20 cu mm) are selected for the Third Harmonic Generation (THG). In this paper, we report the Ti:Sapphire laser at 900 nm and its third harmonic at 300 nm. The desired high ultraviolet (UV) output pulse energy is more than 30 mJ at 300 nm and the energy conversion efficiency from 900 nm to 300 nm is 30%.

Nonlinear Optical Properties of Carotenoid and Chlorophyll Harmonophores

NASA Astrophysics Data System (ADS)

Tokarz, Danielle Barbara

Information regarding the structure and function of living tissues and cells is instrumental to the advancement of cell biology and biophysics. Nonlinear optical microscopy can provide such information, but only certain biological structures generate nonlinear optical signals. Therefore, structural specificity can be achieved by introducing labels for nonlinear optical microscopy. Few studies exist in the literature about labels that facilitate harmonic generation, coined "harmonophores". This thesis consists of the first major investigation of harmonophores for third harmonic generation (THG) microscopy. Carotenoids and chlorophylls were investigated as potential harmonophores. Their nonlinear optical properties were studied by the THG ratio technique. In addition, a tunable refractometer was built in order to determine their second hyperpolarizability (gamma). At 830 nm excitation wavelength, carotenoids and chlorophylls were found to have large negative gamma values however, at 1028 nm, the sign of gamma reversed for carotenoids and remained negative for chlorophylls. Consequently, at 1028 nm wavelength, THG signal is canceled with mixtures of carotenoids and chlorophylls. Furthermore, when such molecules are covalently bonded as dyads or interact within photosynthetic pigment-protein complexes, it is found that additive effects with the gamma values still play a role, however, the overall gamma value is also influenced by the intra-pigment and inter-pigment interaction. The nonlinear optical properties of aggregates containing chlorophylls and carotenoids were the target of subsequent investigations. Carotenoid aggregates were imaged with polarization-dependent second harmonic generation and THG microscopy. Both techniques revealed crystallographic information pertaining to H and J aggregates and beta-carotene crystalline aggregates found in orange carrot. In order to demonstrate THG enhancement due to labeling, cultured cells were labeled with carotenoid incorporated liposomes. In addition, Drosophila melanogaster larvae muscle as well as keratin structures in the hair cortex were labeled with beta-carotene. Polarization-dependent THG studies may be particularly useful in understanding the structural organization that occurs within biological structures containing carotenoids and chlorophylls such as photosynthetic pigment-protein complexes and carotenoid aggregates in plants and alga. Further, artificial labeling with carotenoids and chlorophylls may be useful in clinical applications since they are nontoxic, nutritionally valuable, and they can aid in visualizing structural changes in cellular components.

Cavity enhanced third harmonic generation in graphene

NASA Astrophysics Data System (ADS)

Beckerleg, Chris; Constant, Thomas J.; Zeimpekis, Ioannis; Hornett, Samuel M.; Craig, Chris; Hewak, Daniel W.; Hendry, Euan

2018-01-01

Graphene displays a surprisingly large third order nonlinearity. Here, we report that conversion efficiencies approaching 10-4 are possible for third harmonic generation (THG). Moreover, the atomically thin nature of graphene allows for simple integration in cavity designs to increase this even further. We demonstrate a 117-fold enhancement, of resonant vs non-resonant wavelengths in the THG from graphene due to the integration of a graphene layer with a resonant cavity. This large enhancement occurs as the cavity is resonant for both the fundamental field and the third harmonic. We model this effect using the finite difference time domain approach. By comparing our model with experiment, we are able to deduce the value of a bulk third order susceptibility of graphene of |χ(3)|=4 ×10-17(m/V ) 2 .

Biophotonic endoscopy: a review of clinical research techniques for optical imaging and sensing of early gastrointestinal cancer

PubMed Central

Coda, Sergio; Siersema, Peter D.; Stamp, Gordon W. H.; Thillainayagam, Andrew V.

2015-01-01

Detection, characterization, and staging constitute the fundamental elements in the endoscopic diagnosis of gastrointestinal diseases, but histology still remains the diagnostic gold standard. New developments in endoscopic techniques may challenge histopathology in the near future. An ideal endoscopic technique should combine a wide-field, “red flag” screening technique with an optical contrast or microscopy method for characterization and staging, all simultaneously available during the procedure. In theory, biophotonic advances have the potential to unite these elements to allow in vivo “optical biopsy.” These techniques may ultimately offer the potential to increase the rates of detection of high risk lesions and the ability to target biopsies and resections, and so reduce the need for biopsy, costs, and uncertainty for patients. However, their utility and sensitivity in clinical practice must be evaluated against those of conventional histopathology. This review describes some of the most recent applications of biophotonics in endoscopic optical imaging and metrology, along with their fundamental principles and the clinical experience that has been acquired in their deployment as tools for the endoscopist. Particular emphasis has been placed on translational label-free optical techniques, such as fluorescence spectroscopy, fluorescence lifetime imaging microscopy (FLIM), two-photon and multi-photon microscopy, second harmonic generation (SHG) and third harmonic generation (THG) imaging, optical coherence tomography (OCT), diffuse reflectance, Raman spectroscopy, and molecular imaging. PMID:26528489

Intravital imaging of osteocytes in mouse calvaria using third harmonic generation microscopy

PubMed Central

Cisek, Richard; Wein, Marc N.; Turcotte, Raphaël; Haase, Christa; Yeh, Shu-Chi A.; Bharadwaj, Srinidhi; Raphael, Anthony P.; Paudel, Hari; Alt, Clemens; Liu, Tzu-Ming; Kronenberg, Henry M.; Lin, Charles P.

2017-01-01

Osteocytes are the most abundant cell in the bone, and have multiple functions including mechanosensing and regulation of bone remodeling activities. Since osteocytes are embedded in the bone matrix, their inaccessibility makes in vivo studies problematic. Therefore, a non-invasive technique with high spatial resolution is desired. The purpose of this study is to investigate the use of third harmonic generation (THG) microscopy as a noninvasive technique for high-resolution imaging of the lacunar-canalicular network (LCN) in live mice. By performing THG imaging in combination with two- and three-photon fluorescence microscopy, we show that THG signal is produced from the bone-interstitial fluid boundary of the lacuna, while the interstitial fluid-osteocyte cell boundary shows a weaker THG signal. Canaliculi are also readily visualized by THG imaging, with canaliculi oriented at small angles relative to the optical axis exhibiting stronger signal intensity compared to those oriented perpendicular to the optical axis (parallel to the image plane). By measuring forward- versus epi-detected THG signals in thinned versus thick bone samples ex vivo, we found that the epi-collected THG from the LCN of intact bone contains a superposition of backward-directed and backscattered forward-THG. As an example of a biological application, THG was used as a label-free imaging technique to study structural variations in the LCN of live mice deficient in both histone deacetylase 4 and 5 (HDAC4, HDAC5). Three-dimensional analyses were performed and revealed statistically significant differences between the HDAC4/5 double knockout and wild type mice in the number of osteocytes per volume and the number of canaliculi per lacunar surface area. These changes in osteocyte density and dendritic projections occurred without differences in lacunar size. This study demonstrates that THG microscopy imaging of the LCN in live mice enables quantitative analysis of osteocytes in animal models without the use of dyes or physical sectioning. PMID:29065178

Bi-harmonic cantilever design for improved measurement sensitivity in tapping-mode atomic force microscopy.

PubMed

Loganathan, Muthukumaran; Bristow, Douglas A

2014-04-01

This paper presents a method and cantilever design for improving the mechanical measurement sensitivity in the atomic force microscopy (AFM) tapping mode. The method uses two harmonics in the drive signal to generate a bi-harmonic tapping trajectory. Mathematical analysis demonstrates that the wide-valley bi-harmonic tapping trajectory is as much as 70% more sensitive to changes in the sample topography than the standard single-harmonic trajectory typically used. Although standard AFM cantilevers can be driven in the bi-harmonic tapping trajectory, they require large forcing at the second harmonic. A design is presented for a bi-harmonic cantilever that has a second resonant mode at twice its first resonant mode, thereby capable of generating bi-harmonic trajectories with small forcing signals. Bi-harmonic cantilevers are fabricated by milling a small cantilever on the interior of a standard cantilever probe using a focused ion beam. Bi-harmonic drive signals are derived for standard cantilevers and bi-harmonic cantilevers. Experimental results demonstrate better than 30% improvement in measurement sensitivity using the bi-harmonic cantilever. Images obtained through bi-harmonic tapping exhibit improved sharpness and surface tracking, especially at high scan speeds and low force fields.

Field localization and enhancement of phase-locked second- and third-order harmonic generation in absorbing semiconductor cavities

DOE Office of Scientific and Technical Information (OSTI.GOV)

Roppo, V.; Charles M. Bowden Research Facility, US Army RDECOM, Redstone Arsenal, Alabama 35803; Cojocaru, C.

We predict and experimentally observe the enhancement by three orders of magnitude of phase mismatched second and third harmonic generation in a GaAs cavity at 650 and 433 nm, respectively, well above the absorption edge. Phase locking between the pump and the harmonics changes the effective dispersion of the medium and inhibits absorption. Despite hostile conditions the harmonics resonate inside the cavity and become amplified leading to relatively large conversion efficiencies. Field localization thus plays a pivotal role despite the presence of absorption, and ushers in a new class of semiconductor-based devices in the visible and uv ranges.

Efficient second to ninth harmonic generation using megawatt peak power microchip laser.

PubMed

Bhandari, R; Tsuji, N; Suzuki, T; Nishifuji, M; Taira, T

2013-11-18

We report the design and use of a megawatt peak power Nd:YAG/Cr4+:YAG microchip laser for efficient second to ninth harmonic generation. We show that the sub-nanosecond pulse width region, between 100 ps and 1 ns, is ideally suited for efficient wavelength conversion. Using this feature, we report 85% second harmonic generation efficiency using lithium triborate (LBO), 60% fourth harmonic generation efficiency usingß-barium borate, and 44% IR to UV third harmonic generation efficiency using Type I and Type II LBO. Finally, we report the first demonstration of 118 nm VUV generation in xenon gas using a microchip laser.

Second-harmonic patterned polarization-analyzed reflection confocal microscope

NASA Astrophysics Data System (ADS)

Okoro, Chukwuemeka; Toussaint, Kimani C.

2017-08-01

We introduce the second-harmonic patterned polarization-analyzed reflection confocal (SPPARC) microscope-a multimodal imaging platform that integrates Mueller matrix polarimetry with reflection confocal and second-harmonic generation (SHG) microscopy. SPPARC microscopy provides label-free three-dimensional (3-D), SHG-patterned confocal images that lend themselves to spatially dependent, linear polarimetric analysis for extraction of rich polarization information based on the Mueller calculus. To demonstrate its capabilities, we use SPPARC microscopy to analyze both porcine tendon and ligament samples and find differences in both circular degree-of-polarization and depolarization parameters. Moreover, using the collagen-generated SHG signal as an endogenous counterstain, we show that the technique can be used to provide 3-D polarimetric information of the surrounding extrafibrillar matrix plus cells or EFMC region. The unique characteristics of SPPARC microscopy holds strong potential for it to more accurately and quantitatively describe microstructural changes in collagen-rich samples in three spatial dimensions.

Walk-off reduction, using an external optical plate and Bessel-Gaussian interaction

NASA Astrophysics Data System (ADS)

Masoume, Mansouri; Mohsen, Askarbioki; Saeed Ghavami, Sabouri; Alireza, Khorsandi

2015-02-01

To reduce the walk-off angle of the extraordinary third-harmonic ultraviolet wave at 355 nm generated by type II KTiOPO4 and type I β-BaB2O4 optical crystals, and the Gaussian output beam of a Q-switched Nd:YAG laser, a simple theoretical model was developed based on a rotatable BK7 plate of variable thickness. By rotating the plate up to 35° along the beam direction, we reduced the walk-off angle up to ˜ 13%. The same phenomenon is predicted by the model, confirming the performance of the model. It is found that, due to the walk-off effect, the intensity profile of the third-harmonic generation beam is slightly degraded. To compensate for the observed phenomena and further reduce the walk-off, we used a combination of a convex lens and an axicon to transform the beam profile of the interacting fundamental and second-harmonic generation waves to the zero-order Bessel-Gaussian form. As a result, the walk-off is decreased to ˜48.81 mrad, providing ˜30% relative reduction. By using the same BK7 plate rotated up to 35° along the third-harmonic beam direction, the walk-off angle is further reduced to 38.9 mrad. Moreover, it is observed that the beam profile of the emerged Bessel-Gaussian third-harmonic generation beam remains unchanged with no degradation.

Effects of exciton-plasmon strong coupling on third harmonic generation by two-dimensional WS2 at periodic plasmonic interfaces

NASA Astrophysics Data System (ADS)

Sukharev, Maxim; Pachter, Ruth

2018-03-01

We study theoretically the optical response of a WS2 monolayer located near periodic metal nanostructured arrays in two and three dimensions. The emphasis of the simulations is on the strong coupling between excitons supported by WS2 and surface plasmon-polaritons supported by various periodic plasmonic interfaces. It is demonstrated that a monolayer of WS2 placed in close proximity of periodic arrays of either slits or holes results in a Rabi splitting of the corresponding surface plasmon-polariton resonance as revealed in calculated transmission and reflection spectra. The nonlinear regime, at which the few-layer WS2 exhibits experimentally third harmonic generation (THG), is studied in detail. Monolayer transition metal dichalcogenides (TMDs) do not exhibit THG because they are non-centrosymmetric, but here we use the monolayer as an approximation to a thin TMD nanostructure. We show that in the strong coupling regime the third harmonic signal is significantly affected by plasmon-polaritons and the symmetry of hybrid exciton-plasmon modes. It is also shown that the local electromagnetic field induced by plasmons is the major contributor to the enhancement of the third harmonic signal in three dimensions. The local electromagnetic fields resulting from the third harmonic generation are greatly localized and highly sensitive to the environment, thus making it a great tool for nano-probes.

Shaping the third-harmonic radiation from silicon nanodimers

DOE PAGES

Wang, Lei; Kruk, Sergey; Xu, Lei; ...

2017-01-23

Recent progress in the study of resonant light confinement in high-index dielectric nanostructures suggests a new route for achieving efficient control of both electric and magnetic components of light. It also leads to the enhancement of nonlinear effects near electric and magnetic Mie resonances with an engineered radiation directionality. Furthermore we study the third-harmonic generation from dimers composed of pairs of two identical silicon nanoparticles and demonstrate, both numerically and experimentally, that the multipolar harmonic modes generated by the dimers near the Mie resonances allow the shaping of the directionality of nonlinear radiation.

Single nano-hole as a new effective nonlinear element for third-harmonic generation

NASA Astrophysics Data System (ADS)

Melentiev, P. N.; Konstantinova, T. V.; Afanasiev, A. E.; Kuzin, A. A.; Baturin, A. S.; Tausenev, A. V.; Konyaschenko, A. V.; Balykin, V. I.

2013-07-01

In this letter, we report on a particularly strong optical nonlinearity at the nanometer scale in aluminum. A strong optical nonlinearity of the third order was demonstrated on a single nanoslit. Single nanoslits of different aspect ratio were excited by a laser pulse (120 fs) at the wavelength 1.5 μm, leading predominantly to third-harmonic generation (THG). It has been shown that strong surface plasmon resonance in a nanoslit allows the realization of an effective nanolocalized source of third-harmonic radiation. We show also that a nanoslit in a metal film has a significant advantage in nonlinear processes over its Babinet complementary nanostructure (nanorod): the effective abstraction of heat in a film with a slit makes it possible to use much higher laser radiation intensities.

Time-resolved second-harmonic generation from gold nanoparticle arrays

NASA Astrophysics Data System (ADS)

Ferrara, D. W.; Tetz, K. A.; McMahon, M. D.; Haglund, R. F., Jr.

2007-09-01

We have studied the effects of planar inversion symmetry and particle-coupling of gold nanoparticle (NP) arrays by angle dependent second-harmonic generation (SHG). Time- and angle- resolved measurements were made using a mode-locked Ti:sapphire 800 nm laser onto gold NP arrays with plasmon resonance tuned to match the laser wavelength in order to produce maximum SHG signal. Finite-difference time domain simulations are used to model the near-field distributions for the various geometries and compared to experiment. The arrays were fabricated by focused ion-beam lithography and metal vapor deposition followed by standard lift-off protocols, producing NPs approximately 20nm high with various in-plane dimensions and interparticle gaps. Above a threshold fluence of ~ 7.3 × 10 -5 mJ/cm2 we find that the SHG scales with the third power of intensity, rather than the second, and atomic-force microscopy shows that the NPs have undergone a reshaping process leading to more nearly spherical shapes.

Third-order-harmonic generation in coherently spinning molecules

NASA Astrophysics Data System (ADS)

Prost, E.; Zhang, H.; Hertz, E.; Billard, F.; Lavorel, B.; Bejot, P.; Zyss, Joseph; Averbukh, Ilya Sh.; Faucher, O.

2017-10-01

The rotational Doppler effect occurs when circularly polarized light interacts with a rotating anisotropic material. It is manifested by the appearance of a spectral shift ensuing from the transfer of angular momentum and energy between radiation and matter. Recently, we reported terahertz-range rotational Doppler shifts produced in third-order nonlinear optical conversion [O. Faucher et al., Phys. Rev. A 94, 051402(R) (2016), 10.1103/PhysRevA.94.051402]. The experiment was performed in an ensemble of coherently spinning molecules prepared by a short laser pulse exhibiting a twisted linear polarization. The present work provides an extensive analysis of the rotational Doppler effect in third-order-harmonic generation from spinning linear molecules. The underlying physics is investigated both experimentally and theoretically. The implication of the rotational Doppler effect in higher-order processes like high-order-harmonic generation is discussed.

Determination of collagen fibril size via absolute measurements of second-harmonic generation signals

NASA Astrophysics Data System (ADS)

Bancelin, Stéphane; Aimé, Carole; Gusachenko, Ivan; Kowalczuk, Laura; Latour, Gaël; Coradin, Thibaud; Schanne-Klein, Marie-Claire

2014-09-01

The quantification of collagen fibril size is a major issue for the investigation of pathological disorders associated with structural defects of the extracellular matrix. Second-harmonic generation microscopy is a powerful technique to characterize the macromolecular organization of collagen in unstained biological tissues. Nevertheless, due to the complex coherent building of this nonlinear optical signal, it has never been used to measure fibril diameter so far. Here we report absolute measurements of second-harmonic signals from isolated fibrils down to 30 nm diameter, via implementation of correlative second-harmonic-electron microscopy. Moreover, using analytical and numerical calculations, we demonstrate that the high sensitivity of this technique originates from the parallel alignment of collagen triple helices within fibrils and the subsequent constructive interferences of second-harmonic radiations. Finally, we use these absolute measurements as a calibration for ex vivo quantification of fibril diameter in the Descemet’s membrane of a diabetic rat cornea.

Nonlinear optical effects on the surface of acridine yellow-doped lead-tin fluorophosphate glass

NASA Technical Reports Server (NTRS)

He, K. X.; Bryant, William; Venkateswarlu, Putcha

1991-01-01

The second- and third-order nonlinear optical properties of acridine yellow-doped lead-tin fluorophosphate (LTF) glass have been directly studied by measurement of surface enhanced second harmonic generation and third harmonic generation. The three photon excitation fluorescence is also observed. Based on these results, the large nonlinearities of the acridine LTF system which is a new nonlinear optical material are experimentally demonstrated.

Third-harmonic generation from Mie-type resonances of isolated all-dielectric nanoparticles

NASA Astrophysics Data System (ADS)

Melik-Gaykazyan, Elizaveta V.; Shcherbakov, Maxim R.; Shorokhov, Alexander S.; Staude, Isabelle; Brener, Igal; Neshev, Dragomir N.; Kivshar, Yuri S.; Fedyanin, Andrey A.

2017-03-01

Subwavelength silicon nanoparticles are known to support strongly localized Mie-type modes, including those with resonant electric and magnetic dipolar polarizabilities. Here we compare experimentally the efficiency of the third-harmonic generation from isolated silicon nanodiscs for resonant excitation at the two types of dipolar resonances. Using nonlinear spectroscopy, we observe that the magnetic dipolar mode yields more efficient third-harmonic radiation in contrast to the electric dipolar (ED) mode. This is further supported by full-wave numerical simulations, where the volume-integrated local fields and the directly simulated nonlinear response are shown to be negligible at the ED resonance compared with the magnetic one. This article is part of the themed issue 'New horizons for nanophotonics'.

Bismuth ferrite dielectric nanoparticles excited at telecom wavelengths as multicolor sources by second, third, and fourth harmonic generation.

PubMed

Riporto, Jérémy; Demierre, Alexis; Kilin, Vasyl; Balciunas, Tadas; Schmidt, Cédric; Campargue, Gabriel; Urbain, Mathias; Baltuska, Andrius; Le Dantec, Ronan; Wolf, Jean-Pierre; Mugnier, Yannick; Bonacina, Luigi

2018-05-03

We demonstrate the simultaneous generation of second, third, and fourth harmonics from a single dielectric bismuth ferrite nanoparticle excited using a telecom fiber laser at 1560 nm. We first characterize the signals associated with different nonlinear orders in terms of spectrum, excitation intensity dependence, and relative signal strengths. Successively, on the basis of the polarization-resolved emission curves of the three harmonics, we discuss the interplay of susceptibility tensor components at different orders and show how polarization can be used as an optical handle to control the relative frequency conversion properties.

Formation of 100-nm periodic structures on a titanium surface by exploiting the oxidation and third harmonic generation induced by femtosecond laser pulses.

PubMed

Li, Xian-Feng; Zhang, Cheng-Yun; Li, Hui; Dai, Qiao-Feng; Lan, Sheng; Tie, Shao-Long

2014-11-17

Periodic surface structures with periods as small as about one-tenth of the irradiating femtosecond (fs) laser light wavelength were created on the surface of a titanium (Ti) foil by exploiting laser-induced oxidation and third harmonic generation (THG). They were achieved by using 100-fs laser pulses with a repetition rate of 1 kHz and a wavelength ranging from 1.4 to 2.2 μm. It was revealed that an extremely thin TixOy layer was formed on the surface of the Ti foil after irradiating fs laser light with a fluence smaller than the ablation threshold of Ti, leading to a significant enhancement in THG which may exceed the ablation threshold of TixOy. As compared with Ti, the maximum efficacy factor for TixOy appears at a larger normalized wavevector in the direction perpendicular to the polarization of the fs laser light. As a result, the THG-dominated laser ablation of TixOy induces 100-nm periodic structures parallel to the polarization of the fs laser light. The depth of the periodic structures was found to be ~10 nm by atomic force microscopy and the formation of the thin TixOy layer was verified by energy dispersive X-ray spectroscopy.

Maximum imaging depth comparison in porcine vocal folds using 776-nm vs. 1552-nm excitation wavelengths

NASA Astrophysics Data System (ADS)

Yildirim, Murat; Ferhanoglu, Onur; Kobler, James B.; Zeitels, Steven M.; Ben-Yakar, Adela

2013-02-01

Vocal fold scarring is one of the major causes of voice disorders and may arise from overuse or post-surgical wound healing. One promising treatment utilizes the injection of soft biomaterials aimed at restoring viscoelasticity of the outermost vibratory layer of the vocal fold, superficial lamina propria (SLP). However, the density of the tissue and the required injection pressure impair proper localization of the injected biomaterial in SLP. To enhance treatment effectiveness, we are investigating a technique to image and ablate sub-epithelial planar voids in vocal folds using ultrafast laser pulses to better localize the injected biomaterial. It is challenging to optimize the excitation wavelength to perform imaging and ablation at depths suitable for clinical use. Here, we compare maximum imaging depth using two photon autofluorescence and second harmonic generation with third-harmonic generation imaging modalities for healthy porcine vocal folds. We used a home-built inverted nonlinear scanning microscope together with a high repetition rate (2 MHz) ultrafast fiber laser (Raydiance Inc.). We acquired both two-photon autofluorescence and second harmonic generation signals using 776 nm wavelength and third harmonic generation signals using 1552 nm excitation wavelength. We observed that maximum imaging depth with 776 nm wavelength is significantly improved from 114 μm to 205 μm when third harmonic generation is employed using 1552 nm wavelength, without any observable damage in the tissue.

Image storage in coumarin-based copolymer thin films by photoinduced dimerization.

PubMed

Gindre, Denis; Iliopoulos, Konstantinos; Krupka, Oksana; Champigny, Emilie; Morille, Yohann; Sallé, Marc

2013-11-15

We report a technique to encode grayscale digital images in thin films composed of copolymers containing coumarins. A nonlinear microscopy setup was implemented and two nonlinear optical processes were used to store and read information. A third-order process (two-photon absorption) was used to photoinduce a controlled dimer-to-monomer ratio within a defined tiny volume in the material, which corresponds to each recorded bit of data. Moreover, a second-order process (second-harmonic generation) was used to read the stored information, which has been found to be highly dependent upon the monomer-to-dimer ratio.

Measuring the molecular second hyperpolarizability in absorptive solutions by the third harmonic generation ratio technique.

PubMed

Tokarz, Danielle; Cisek, Richard; Prent, Nicole; Fekl, Ulrich; Barzda, Virginijus

2012-11-28

Measurement of the second hyperpolarizability (γ) values of compounds can provide insight into the molecular structural requirements for enhancement of third harmonic generation (THG) signal. A convenient method for measuring the γ of compounds in solutions was developed by implementing the THG ratio method which is based on measuring the THG intensity from two interfaces using a nonlinear optical microscope while accounting for the refractive index of solutions at the fundamental and third harmonic wavelengths. We demonstrated that the difference in refractive index at both wavelengths strongly influenced the calculation of γ values when compounds have absorption near the third harmonic or fundamental wavelength. To this end, a refractometer with the wavelength tuning range from UV to near IR was constructed, and the measured refractive indices were used to extract the γ values. The γ values of carotenoids and chlorophylls found in photosynthetic pigment-protein complexes were explored. Large differences in the refractive index at third harmonic and fundamental wavelengths for chlorophylls result in γ values that are more than two orders of magnitude larger than γ values for carotenoids as well as the sign of chlorophylls'γ values is negative while carotenoids have positive γ values. Copyright © 2012 Elsevier B.V. All rights reserved.

Layer-Dependent Third-Harmonic Generation in Graphene

NASA Astrophysics Data System (ADS)

Yang, Hao; Guan, Honghua; Dadap, Jerry; Osgood, Richard; Richard Osgood Team

Graphene has become a subject of intense interest and study because of its remarkable 2D electronic properties. Multilayer graphene also offers an array of properties that are also of interest for optical physics and devices. Despite its second-order-nonlinear optical response is intrinsically weak, third-order nonlinear optical effects in graphene are symmetry-allowed thus leading to studies of several third-order process in few-layer graphene. In this work, we report third-harmonic generation in multilayer graphene mounted on fused silica and with thicknesses which approach the bulk continuum. THG signals show cubic power dependence with respect to the intensity of fundamental beam. Third-harmonic generation spectroscopy enables a good fit using linear optical detection, which shows strong contrast for different layer number graphene. The maximum THG efficiency appears at layer number around 30. Two models are used for describing this layer dependent phenomenon and shows absorption plays a key role in THG of multilayer graphene. This work also provides a new imaging technology for graphene detection and identification with better contrast and resolution. U.S. Department of Energy under Contract No. DE-FG 02-04-ER-46157.

Characteristics of a KA-band third-harmonic peniotron driven by a high-quality linear axis-encircling electron beam

NASA Astrophysics Data System (ADS)

Zhao, Xiaoyun; Tuo, Xianguo; Ge, Qing; Peng, Ying

2017-12-01

We employ a high-quality linear axis-encircling electron beam generated by a Cuccia coupler to drive a Ka-band third-harmonic peniotron and develop a self-consistent nonlinear calculation code to numerically analyze the characteristics of the designed peniotron. It is demonstrated that through a Cuccia coupler, a 6 kV, 0.5 A pencil beam and an input microwave power of 16 kW at 10 GHz can generate a 37 kV, 0.5 A linear axis-encircling beam, and it is characterized by a very low velocity spread. Moreover, the electron beam guiding center deviation can be adjusted easily. Driven by such a beam, a 30 GHz, Ka-band third-harmonic peniotron is predicted to achieve a conversion efficiency of 51.0% and a microwave output power of 9.44 kW; the results are in good agreement with the Magic3D simulation. Using this code, we studied the factors influencing the peniotron performance, and it can provide some guidelines for the design of a Ka-band third-harmonic peniotron driven by a linear electron beam and can promote the application of high-harmonic peniotrons in practice.

Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy.

PubMed

You, Sixian; Tu, Haohua; Chaney, Eric J; Sun, Yi; Zhao, Youbo; Bower, Andrew J; Liu, Yuan-Zhi; Marjanovic, Marina; Sinha, Saurabh; Pu, Yang; Boppart, Stephen A

2018-05-29

Intravital microscopy (IVM) emerged and matured as a powerful tool for elucidating pathways in biological processes. Although label-free multiphoton IVM is attractive for its non-perturbative nature, its wide application has been hindered, mostly due to the limited contrast of each imaging modality and the challenge to integrate them. Here we introduce simultaneous label-free autofluorescence-multiharmonic (SLAM) microscopy, a single-excitation source nonlinear imaging platform that uses a custom-designed excitation window at 1110 nm and shaped ultrafast pulses at 10 MHz to enable fast (2-orders-of-magnitude improvement), simultaneous, and efficient acquisition of autofluorescence (FAD and NADH) and second/third harmonic generation from a wide array of cellular and extracellular components (e.g., tumor cells, immune cells, vesicles, and vessels) in living tissue using only 14 mW for extended time-lapse investigations. Our work demonstrates the versatility and efficiency of SLAM microscopy for tracking cellular events in vivo, and is a major enabling advance in label-free IVM.

Second and third order nonlinear optical properties of conjugated molecules and polymers

NASA Technical Reports Server (NTRS)

Perry, Joseph W.; Stiegman, Albert E.; Marder, Seth R.; Coulter, Daniel R.; Beratan, David N.; Brinza, David E.

1988-01-01

Second- and third-order nonlinear optical properties of some newly synthesized organic molecules and polymers are reported. Powder second-harmonic-generation efficiencies of up to 200 times urea have been realized for asymmetric donor-acceptor acetylenes. Third harmonic generation chi(3)s have been determined for a series of small conjugated molecules in solution. THG chi(3)s have also been determined for a series of soluble conjugated copolymers prepared using ring-opening metathesis polymerization. The results are discussed in terms of relevant molecular and/or macroscopic structural features of these conjugated organic materials.

Chi 3 dispersion in planar tantalum pentoxide waveguides in the telecommunications window.

PubMed

Chen, Ruiqi Y; Charlton, Martin D B; Lagoudakis, Pavlos G

2009-04-01

We report on the dispersion of the third-order nonlinear susceptibility (chi(3) or "Chi 3") in planar Ta2O5 waveguides in the telecommunications spectral window. We utilize the observation of third-harmonic generation under ultrashort pulsed excitation as a reference-free characterization method of chi(3) and obtain a large nonlinear coefficient, 2x10(-13) esu, at 1550 nm. Our observation of efficient third-harmonic generation in Ta2O5 waveguides in the telecoms window reveals the potential of this material system in high-speed integrated nonlinear optical switches.

Analysis of Even Harmonics Generation in an Isolated Electric Power System

NASA Astrophysics Data System (ADS)

Kanao, Norikazu; Hayashi, Yasuhiro; Matsuki, Junya

Harmonics bred from loads are mainly odd order because the current waveform has half-wave symmetry. Since the even harmonics are negligibly small, those are not generally measured in electric power systems. However, even harmonics were measured at a 500/275/154kV substation in Hokuriku Electric Power Company after removal of a transmission line fault. The even harmonics caused malfunctions of protective digital relays because the relays used 4th harmonics at the input filter as automatic supervisory signal. This paper describes the mechanism of generation of the even harmonics by comparing measured waveforms with ATP-EMTP simulation results. As a result of analysis, it is cleared that even harmonics are generated by three causes. The first cause is a magnetizing current of transformers due to flux deviation by DC component of a fault current. The second one is due to harmonic conversion of a synchronous machine which generates even harmonics when direct current component or even harmonic current flow into the machine. The third one is that increase of harmonic impedance due to an isolated power system produces harmonic voltages. The design of the input filter of protective digital relays should consider even harmonics generation in an isolated power system.

Optical coherence tomography and non-linear microscopy for paintings - a study of the complementary capabilities and laser degradation effects.

PubMed

Liang, Haida; Mari, Meropi; Cheung, Chi Shing; Kogou, Sotiria; Johnson, Phillip; Filippidis, George

2017-08-07

This paper examines for the first time the potential complementary imaging capabilities of Optical coherence tomography (OCT) and non-linear microscopy (NLM) for multi-modal 3D examination of paintings following the successful application of OCT to the in situ, non-invasive examination of varnish and paint stratigraphy of historic paintings and the promising initial studies of NLM of varnish samples. OCT provides image contrast through the optical scattering and absorption properties of materials, while NLM provides molecular information through multi-photon fluorescence and higher harmonics generation (second and third harmonic generation). OCT is well-established in the in situ non-invasive imaging of the stratigraphy of varnish and paint layers. While NLM examination of transparent samples such as fresh varnish and some transparent paints showed promising results, the ultimate use of NLM on paintings is limited owing to the laser degradation effects caused by the high peak intensity of the laser source necessary for the generation of non-linear phenomena. The high intensity normally employed in NLM is found to be damaging to all non-transparent painting materials from slightly scattering degraded varnish to slightly absorbing paint at the wavelength of the laser excitation source. The results of this paper are potentially applicable to a wide range of materials given the diversity of the materials encountered in paintings (e.g. minerals, plants, insects, oil, egg, synthetic and natural varnish).

Automatic computation and solution of generalized harmonic balance equations

NASA Astrophysics Data System (ADS)

Peyton Jones, J. C.; Yaser, K. S. A.; Stevenson, J.

2018-02-01

Generalized methods are presented for generating and solving the harmonic balance equations for a broad class of nonlinear differential or difference equations and for a general set of harmonics chosen by the user. In particular, a new algorithm for automatically generating the Jacobian of the balance equations enables efficient solution of these equations using continuation methods. Efficient numeric validation techniques are also presented, and the combined algorithm is applied to the analysis of dc, fundamental, second and third harmonic response of a nonlinear automotive damper.

Harmonics Generation by Surface Plasmon Polaritons on Single Nanowires.

PubMed

de Hoogh, Anouk; Opheij, Aron; Wulf, Matthias; Rotenberg, Nir; Kuipers, L

2016-08-17

We present experimental observations of visible wavelength second- and third-harmonic generation on single plasmonic nanowires of variable widths. We identify that near-infrared surface plasmon polaritons, which are guided along the nanowire, act as the source of the harmonics generation. We discuss the underlying mechanism of this nonlinear process, using a combination of spatially resolved measurements and numerical simulations to show that the visible harmonics are generated via a combination of both local and propagating plasmonic modes. Our results provide the first demonstration of nanoscale nonlinear optics with guided, propagating plasmonic modes on a lithographically defined chip, opening up new routes toward integrated optical circuits for information processing.

In vivo polarization dependant Second and Third harmonic generation imaging of Caenorhabditis elegans pharyngeal muscles

NASA Astrophysics Data System (ADS)

Filippidis, G.; Troulinaki, K.; Fotakis, C.; Tavernarakis, N.

2009-07-01

In this study Second and Third harmonic generation (SHG-THG) imaging measurements were performed to the pharyngeal muscles of the nematode Caenorhabditis elegans, in vivo with linearly polarized laser beam. Complementary information about the anatomy of the pharynx and the morphology of the anterior part of the worm were extracted. THG signals proved to have no dependence on incident light polarization, while SHG images are highly sensitive to the changes of the incident linearly polarized light.

Noninvasive intravital cellular diagnosis of atopic dermatitis by using harmonic optical virtual biopsy

NASA Astrophysics Data System (ADS)

Chen, Szu-Yu; Lee, Jyh-Hong; Chiang, Bor-Luen; Sun, Chi-Kuang

2007-02-01

Atopic dermatitis (AD) is now very common in people who live in cities, especially for babies and children. Since the cause of AD is still not completely understood and each person may have his own mixed symptoms that can change over time, diagnosis of AD can not be done precisely. Unlike some skin diseases, physical biopsy is rarely used in diagnosing AD on account of its low urgency. Thus, only indirect diagnoses, like asking for a medical history to learn about the symptoms and to rule out other diseases can be carried out. To gain insight into cellular details of AD for long-term diagnosing without physical biopsy, a noninvasive in vivo tool with a sub-micron subsurface resolution and high penetrability has to be used. In this presentation, we show that harmonic optical virtual biopsy can provide the required noninvasive cellular imaging, and is ideal for future clinical diagnosis of AD. Harmonic optical microscopy has been demonstrated to have the capability to reveal cellular morphology of human skin from epidermis to dermis layer. Third harmonic generation (THG), which is sensitive to inhomogeneous interfaces, can show the structures of skins, and can be used to reveal the morphological changes, for example, the thicken cuticle which is a common symptom of AD. Second harmonic generation (SHG), which occurs in non-centrosymmetric structures, has excellent contrast in collagen fibers and can show the pathological changes of dermis layer. Utilizing both THG and SHG, useful information may be given to facilitate the diagnosis of AD.

Epi-detected quadruple-modal nonlinear optical microscopy for label-free imaging of the tooth

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Zi; Zheng, Wei; Huang, Zhiwei, E-mail: biehzw@nus.edu.sg

2015-01-19

We present an epi-detected quadruple-modal nonlinear optical microscopic imaging technique (i.e., coherent anti-Stokes Raman scattering (CARS), second-harmonic generation (SHG), third-harmonic generation (THG), and two-photon excited fluorescence (TPEF)) based on a picosecond (ps) laser-pumped optical parametric oscillator system for label-free imaging of the tooth. We demonstrate that high contrast ps-CARS images covering both the fingerprint (500–1800 cm{sup −1}) and high-wavenumber (2500–3800 cm{sup −1}) regions can be acquired to uncover the distributions of mineral and organic biomaterials in the tooth, while high quality TPEF, SHG, and THG images of the tooth can also be acquired under ps laser excitation without damaging the samples. Themore » quadruple-modal nonlinear microscopic images (CARS/SHG/THG/TPEF) acquired provide better understanding of morphological structures and biochemical/biomolecular distributions in the dentin, enamel, and the dentin-enamel junction of the tooth without labeling, facilitating optical diagnosis and characterization of the tooth in dentistry.« less

Third harmonic generation imaging of intact human cerebral organoids to assess key components of early neurogenesis in Rett Syndrome (Conference Presentation)

NASA Astrophysics Data System (ADS)

Yildirim, Murat; Feldman, Danielle; Wang, Tianyu; Ouzounov, Dimitre G.; Chou, Stephanie; Swaney, Justin; Chung, Kwanghun; Xu, Chris; So, Peter T. C.; Sur, Mriganka

2017-02-01

Rett Syndrome (RTT) is a pervasive, X-linked neurodevelopmental disorder that predominantly affects girls. It is mostly caused by a sporadic mutation in the gene encoding methyl CpG-binding protein 2 (MeCP2).The clinical features of RTT are most commonly reported to emerge between the ages of 6-18 months and implicating RTT as a disorder of postnatal development. However, a variety of recent evidence from our lab and others demonstrates that RTT phenotypes are present at the earliest stages of brain development including neurogenesis, migration, and patterning in addition to stages of synaptic and circuit development and plasticity. We have used RTT patient-derived induced pluripotent stem cells to generate 3D human cerebral organoids that can serve as a model for human neurogenesis in vitro. We aim to expand on our existing findings in order to determine aberrancies at individual stages of neurogenesis by performing structural and immunocytochemical staining in isogenic control and MeCP2-deficient organoids. In addition, we aim to use Third Harmonic Generation (THG) microscopy as a label-free, nondestructive 3D tissue visualization method in order to gain a complete understanding of the structural complexity that underlies human neurogenesis. As a proof of concept, we have performed THG imaging in healthy intact human cerebral organoids cleared with SWITCH. We acquired an intrinsic THG signal with the following laser configurations: 400 kHz repetition rate, 65 fs pulse width laser at 1350 nm wavelength. In these THG images, nuclei are clearly delineated and cross sections demonstrate the depth penetration capacity (< 1mm) that extends throughout the organoid. Imaging control and MeCP2-deficient human cerebral organoids in 2D sections reveals structural and protein expression-based alterations that we expect will be clearly elucidated via both THG and three-photon fluorescence microscopy.

Bound States and the Third Harmonic Generation in an Electric Field Biased Semi-parabolic Quantum Well

NASA Astrophysics Data System (ADS)

Zhang, Li; Xie, Hong-Jing

2003-11-01

Within the framework of the compact density matrix approach, the third-harmonic generation (THG) in an electric-field-biased semi-parabolic quantum well (QW) has been deduced and investigated. Via variant of displacement harmonic oscillation, the exact electronic states in the semi-parabolic QW with an applied electric field have also been obtained and discussed. Numerical results on typical GaAs material reveal that, electric fields and confined potential frequency of semi-parabolic QW have obvious influences on the energy levels of electronic states and the THG in the semi-parabolic QW systems. The project supported in part by Guangdong Provincial Natural Science Foundation of China

Magnetization-induced second- and third-harmonic generation in transparent magnetic films

NASA Astrophysics Data System (ADS)

Ohkoshi, Shin-Ichi; Shimura, Jusuke; Ikeda, Katsuyoshi; Hashimoto, Kazuhito

2005-01-01

We describe the magnetization-induced second-harmonic (SH) generation in (FeIIxCrII1-x)1.5[CrIII(CN)6]. 7.5H2O and the magnetization-induced third-harmonic (TH) generation in Y1.5Bi1.5Fe3.8Al1.2O12 (Bi, Al:YIG). The polarization plane of a SH wave from a (FeIIxCrII1-x)1.5[CrIII(CN)6].7.5H2O film was rotated by an applied external magnetic field. This SH rotation is ascribed to the interaction between the electric polarization along the out-of-plane and spontaneous magnetizations. In particular, the magnetic linear term χijkLmagn(1) contributed to the SH rotation. Applying a longitudinal external magnetic field to a Bi,Al:YIG magnetic film rotated the polarization plane of the TH wave. This TH rotation is understood by the contribution of the magnetic term of χyxxxZmagn(1) in a third-order nonlinear optical susceptibility.

Second and third harmonic generation associated to infrared transitions in a Morse quantum well under applied electric and magnetic fields

NASA Astrophysics Data System (ADS)

Restrepo, R. L.; Kasapoglu, E.; Sakiroglu, S.; Ungan, F.; Morales, A. L.; Duque, C. A.

2017-09-01

The effects of electric and magnetic fields on the second and third harmonic generation coefficients in a Morse potential quantum well are theoretically studied. The energy levels and corresponding wave functions are obtained by solving the Schrödinger equation for the electron in the parabolic band scheme and effective mass approximations and the envelope function approach. The results show that both the electric and the magnetic fields have significant influence on the magnitudes and resonant peak energy positions of the second and third harmonic generation responses. In general, the Morse potential profile becomes wider and shallower as γ -parameter increases and so the energies of the bound states will be functions of this parameter. Therefore, we can conclude that the effects of the electric and magnetic fields can be used to tune and control the optical properties of interest in the range of the infrared electromagnetic spectrum.

On the possible origin of bulk third harmonic generation in skin cells

NASA Astrophysics Data System (ADS)

Su, Tung-Yu; Liao, Chien-Sheng; Yang, Chih-Yuan; Zhuo, Guan-Yu; Chen, Szu-Yu; Chu, Shi-Wei

2011-09-01

We studied third harmonic generation (THG) of melanin solution with concentrations similar to melanocytes in human skin. In contrast to conventional observation of THG at interface, bulk THG was detected inside the solution due to the formation of melanin hydrocolloids. A linear relationship between melanin concentration and THG intensity was found, suggesting THG originated from high-order hyper-Rayleigh scattering. By fitting this linear relationship, third-order hyperpolarizability of melanin hydrocolloids was determined to be three orders larger than that of water. Our result will be useful for interpretation of THG signals in skin and other tissues containing colloidal particles.

Study of third order nonlinearity of chalcogenide thin films using third harmonic generation measurements

NASA Astrophysics Data System (ADS)

Rani, Sunita; Mohan, Devendra; Kumar, Manish; Sanjay

2018-05-01

Third order nonlinear susceptibility of (GeSe3.5)100-xBix (x = 0, 10, 14) and ZnxSySe100-x-y (x = 2, y = 28; x = 4, y = 20; x = 6, y = 12; x = 8, y = 4) amorphous chalcogenide thin films prepared using thermal evaporation technique is estimated. The dielectric constant at incident and third harmonic wavelength is calculated using "PARAV" computer program. 1064 nm wavelength of Nd: YAG laser is incident on thin film and third harmonic signal at 355 nm wavelength alongwith fundamental light is obtained in reflection that is separated from 1064 nm using suitable optical filter. Reflected third harmonic signal is measured to trace the influence of Bi and Zn on third order nonlinear susceptibility and is found to increase with increase in Bi and Zn content in (GeSe3.5)100-xBix, and ZnxSySe100-x-y chalcogenide thin films respectively. The excellent optical nonlinear property shows the use of chalcogenide thin films in photonics for wavelength conversion and optical data processing.

Third harmonic from air breakdown plasma induced by nanosecond laser pulses

NASA Astrophysics Data System (ADS)

Stafe, M.; Negutu, C.; Puscas, N. N.

2018-06-01

Harmonic generation is a nonlinear optical effect consisting in frequency up-conversion of intense laser radiation when phase-matching conditions are fulfilled. Here, we study the mechanisms involved in the third harmonic (TH) generation process, the conversion efficiency, and the properties of TH radiation generated in air by focusing infrared linearly polarized nanosecond laser pulses at intensities of the order of TW/cm2. By analyzing the emission from the air breakdown plasma, we demonstrate that filamentary breakdown plasma containing molecular nitrogen ions acts as an optical nonlinear medium enabling generation of TH radiation in the axial direction. The data reveal important properties of the TH radiation: maximum conversion efficiency of 0.04%, sinc2 dependence of the TH intensity on the square root of the pump intensity, and three times smaller divergence and pulse duration of TH as compared to the pump radiation.

Quantitative analysis on collagen of dermatofibrosarcoma protuberans skin by second harmonic generation microscopy.

PubMed

Wu, Shulian; Huang, Yudian; Li, Hui; Wang, Yunxia; Zhang, Xiaoman

2015-01-01

Dermatofibrosarcoma protuberans (DFSP) is a skin cancer usually mistaken as other benign tumors. Abnormal DFSP resection results in tumor recurrence. Quantitative characterization of collagen alteration on the skin tumor is essential for developing a diagnostic technique. In this study, second harmonic generation (SHG) microscopy was performed to obtain images of the human DFSP skin and normal skin. Subsequently, structure and texture analysis methods were applied to determine the differences in skin texture characteristics between the two skin types, and the link between collagen alteration and tumor was established. Results suggest that combining SHG microscopy and texture analysis methods is a feasible and effective method to describe the characteristics of skin tumor like DFSP. © Wiley Periodicals, Inc.

Macromolecular structure of cellulose studied by second-harmonic generation imaging microscopy

NASA Astrophysics Data System (ADS)

Brown, R. Malcom; Millard, Andrew C.; Campagnola, Paul J.

2003-11-01

The macromolecular structure of purified cellulose samples is studied by second-harmonic generation (SHG) imaging microscopy. We show that the SHG contrast in both Valonia and Acetobacter cellulose strongly resembles that of collagen from animal tissues, both in terms of morphology and polarization anisotropy. Polarization analysis shows that microfibrils in each lamella are highly aligned and ordered and change directions by 90° in adjacent lamellae. The angular dependence of the SHG intensity fits well to a cos2 θ distribution, which is characteristic of the electric dipole interaction. Enzymatic degradation of Valonia fibers by cellulase is followed in real time by SHG imaging and results in exponential decay kinetics, showing that SHG imaging microscopy is ideal for monitoring dynamics in biological systems.

Towards a Compact Fiber Laser for Multimodal Imaging

NASA Astrophysics Data System (ADS)

Nie, Bai; Saytashev, Ilyas; Dantus, Marcos

We report on multimodal depth-resolved imaging of unstained living Drosophila Melanogaster larva using sub-50 fs pulses centered at 1060 nm wavelength. Both second harmonic and third harmonic generation imaging modalities are demonstrated.

Towards a compact fiber laser for multimodal imaging

NASA Astrophysics Data System (ADS)

Nie, Bai; Saytashev, Ilyas; Dantus, Marcos

2014-03-01

We report on multimodal depth-resolved imaging of unstained living Drosophila Melanogaster larva using sub-50 fs pulses centered at 1060 nm wavelength. Both second harmonic and third harmonic generation imaging modalities are demonstrated.

Second- and third-harmonic generation in metal-based structures

DOE Office of Scientific and Technical Information (OSTI.GOV)

Scalora, M.; Akozbek, N.; Bloemer, M. J.

We present a theoretical approach to the study of second- and third-harmonic generation from metallic structures and nanocavities filled with a nonlinear material in the ultrashort pulse regime. We model the metal as a two-component medium, using the hydrodynamic model to describe free electrons and Lorentz oscillators to account for core electron contributions to both the linear dielectric constant and harmonic generation. The active nonlinear medium that may fill a metallic nanocavity, or be positioned between metallic layers in a stack, is also modeled using Lorentz oscillators and surface phenomena due to symmetry breaking are taken into account. We studymore » the effects of incident TE- and TM-polarized fields and show that a simple reexamination of the basic equations reveals additional, exploitable dynamical features of nonlinear frequency conversion in plasmonic nanostructures.« less

Enhanced third harmonic generation from the epsilon-near-zero modes of ultrathin films

DOE Office of Scientific and Technical Information (OSTI.GOV)

Luk, Ting S.; De Ceglia, Domenico; Liu, Sheng

We demonstrate, through our experimentation, efficient third harmonic generation from an indium tin oxide nanofilm (λ/42 thick) on a glass substrate for a pump wavelength of 1.4 μm. A conversion efficiency of 3.3 × 10 -6 is achieved by exploiting the field enhancement properties of the epsilon-near-zero mode with an enhancement factor of 200. Furthermore, this nanoscale frequency conversion method is applicable to other plasmonic materials and reststrahlen materials in proximity of the longitudinal optical phonon frequencies.

Enhanced third harmonic generation from the epsilon-near-zero modes of ultrathin films

DOE Office of Scientific and Technical Information (OSTI.GOV)

Luk, Ting S., E-mail: tsluk@sandia.gov; Liu, Sheng; Campione, Salvatore

We experimentally demonstrate efficient third harmonic generation from an indium tin oxide nanofilm (λ/42 thick) on a glass substrate for a pump wavelength of 1.4 μm. A conversion efficiency of 3.3 × 10{sup −6} is achieved by exploiting the field enhancement properties of the epsilon-near-zero mode with an enhancement factor of 200. This nanoscale frequency conversion method is applicable to other plasmonic materials and reststrahlen materials in proximity of the longitudinal optical phonon frequencies.

PREFACE: Ultrafast biophotonics Ultrafast biophotonics

NASA Astrophysics Data System (ADS)

Gu, Min; Reid, Derryck; Ben-Yakar, Adela

2010-08-01

The use of light to explore biology can be traced to the first observations of tissue made with early microscopes in the mid-seventeenth century, and has today evolved into the discipline which we now know as biophotonics. This field encompasses a diverse range of activities, each of which shares the common theme of exploiting the interaction of light with biological material. With the rapid advancement of ultrafast optical technologies over the last few decades, ultrafast lasers have increasingly found applications in biophotonics, to the extent that the distinctive new field of ultrafast biophotonics has now emerged, where robust turnkey ultrafast laser systems are facilitating cutting-edge studies in the life sciences to take place in everyday laboratories. The broad spectral bandwidths, precision timing resolution, low coherence and high peak powers of ultrafast optical pulses provide unique opportunities for imaging and manipulating biological systems. Time-resolved studies of bio-molecular dynamics exploit the short pulse durations from such lasers, while other applications such as optical coherence tomography benefit from the broad optical bandwidths possible by using super-continuum generation and additionally allowing for high speed imaging with speeds as high as 47 000 scans per second. Continuing progress in laser-system technology is accelerating the adoption of ultrafast techniques across the life sciences, both in research laboratories and in clinical applications, such as laser-assisted in situ keratomileusis (LASIK) eye surgery. Revolutionizing the field of optical microscopy, two-photon excitation fluorescence (TPEF) microscopy has enabled higher spatial resolution with improved depth penetration into biological specimens. Advantages of this nonlinear optical process include: reduced photo-interactions, allowing for extensive imaging time periods; simultaneously exciting multiple fluorescent molecules with only one excitation wavelength; and reduced chromatic aberration effects. These extensive advantages have led to further exploration of nonlinear processes including second-harmonic generation (SHG) microscopy and third-harmonic generation (THG) microscopy. Second-harmonic generation has provided biologists with an extremely powerful tool for generating contrast in biological imaging, with the additional benefit of non-invasive three-dimensional imaging. The recent popularity of THG microscopy is largely due to the fact that three-dimensional imaging is achievable without the need for any labels, but rather relying on the intrinsic properties of the biological specimen itself. This optical nonlinear technique has attracted much attention recently from the biological community due to its non-invasive capabilities. Users of ultrafast lasers in the biological and medical fields are becoming a fast-growing community, employing pulse-shaping microscopy, resolution-enhancing microscopy techniques, linear and nonlinear micro-spectroscopy, functional deep-tissue imaging, optical coherence tomography, nonlinear fluorescence microscopy, molecular imaging and control, harmonic microscopy and femtosecond lifetime imaging, for cutting-edge research concerning the interaction of light with biological dynamics. The adaptability of ultrafast lasers to interact with a large array of materials through nonlinear excitation has enabled precise control of laser fluence allowing for highly localized material interactions, permitting micro-structured fabricated surfaces. The resultant multi-dimensional fabricated micro-structures are capable of replicating and/or manipulating microenvironments for controlled cell biology. In this special issue of Journal of Optics readers have a chance to view a collection of new contributions to the growing research field of ultrafast biophotonics. They are presented with recent advances in ultrafast technology applied to biological and medical investigations, where topics include advances in the visualization and identification of photo-reaction dynamics of biological functions under relevant physiological conditions, theoretically proposed imaging designs for obtaining super-resolved optical sectioned images in single exposures and fabricated micro-structured surfaces for biological micro-environments. We hope the collection will stimulate innovative new research in this growing field by showcasing new techniques for the visualization and manipulation of complex biological systems using linear and and nonlinear optical processes. Professor Min Gu would like to acknowledge Dr Betty Kouskousis for her contribution and support towards this editorial.

Third harmonic frequency generation by type-I critically phase-matched LiB3O5 crystal by means of optically active quartz crystal.

PubMed

Gapontsev, Valentin P; Tyrtyshnyy, Valentin A; Vershinin, Oleg I; Davydov, Boris L; Oulianov, Dmitri A

2013-02-11

We present a method of third harmonic generation at 355 nm by frequency mixing of fundamental and second harmonic radiation of an ytterbium nanosecond pulsed all-fiber laser in a type-I phase-matched LiB(3)O(5) (LBO) crystal where originally orthogonal polarization planes of the fundamental and second harmonic beams are aligned by an optically active quartz crystal. 8 W of ultraviolet light at 355 nm were achieved with 40% conversion efficiency from 1064 nm radiation. The conversion efficiency obtained in a type-I phase-matched LBO THG crystal was 1.6 times higher than the one achieved in a type-II LBO crystal at similar experimental conditions. In comparison to half-wave plates traditionally used for polarization alignment the optically active quartz crystal has much lower temperature dependence and requires simpler optical alignment.

Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues.

PubMed Central

Campagnola, Paul J; Millard, Andrew C; Terasaki, Mark; Hoppe, Pamela E; Malone, Christian J; Mohler, William A

2002-01-01

We find that several key endogenous protein structures give rise to intense second-harmonic generation (SHG)-nonabsorptive frequency doubling of an excitation laser line. Second-harmonic imaging microscopy (SHIM) on a laser-scanning system proves, therefore, to be a powerful and unique tool for high-resolution, high-contrast, three-dimensional studies of live cell and tissue architecture. Unlike fluorescence, SHG suffers no inherent photobleaching or toxicity and does not require exogenous labels. Unlike polarization microscopy, SHIM provides intrinsic confocality and deep sectioning in complex tissues. In this study, we demonstrate the clarity of SHIM optical sectioning within unfixed, unstained thick specimens. SHIM and two-photon excited fluorescence (TPEF) were combined in a dual-mode nonlinear microscopy to elucidate the molecular sources of SHG in live cells and tissues. SHG arose not only from coiled-coil complexes within connective tissues and muscle thick filaments, but also from microtubule arrays within interphase and mitotic cells. Both polarization dependence and a local symmetry cancellation effect of SHG allowed the signal from species generating the second harmonic to be decoded, by ratiometric correlation with TPEF, to yield information on local structure below optical resolution. The physical origin of SHG within these tissues is addressed and is attributed to the laser interaction with dipolar protein structures that is enhanced by the intrinsic chirality of the protein helices. PMID:11751336

Multi-Photon Micro-Spectroscopy of Biological Specimens

DTIC Science & Technology

2000-07-01

Micro-spectroscopy, multi-photon fluorescence spectroscopy, second harmonic generation, plant tissues, stem, chloroplast, protoplast, maize, Arabidopsis...harmonic generation (SHG) in the plant cell 5wall. In this case, micro-spectroscopy provides a means of verification that, indeed, SHG occurs in plant ...fluorescence microscopy -the response of plant cells to high intensity illumination," Micron (in press) 2000. 3. H.-C. Huang and C. -C Chen, "Genome

Polarization-resolved second-harmonic-generation imaging of photoaged dermal collagen fiber

NASA Astrophysics Data System (ADS)

Yasui, Takeshi; Takahashi, Yu; Araki, Tsutomu

2009-02-01

Polarization-resolved second-harmonic-generation (SHG) microscopy is useful for assessment of collagen fiber orientation in tissues. In this paper, we investigated the relation between wrinkle direction and collagen orientation in ultraviolet-B-exposed (UVB-exposed) skin using polarization-resolved SHG microscopy. A polarization anisotropic image of the SHG light indicated that wrinkle direction in UVB-exposed skin is predominantly parallel to the orientation of dermal collagen fibers whereas no-UVB-exposed skin was dominated by collagen orientation parallel to the meridian line of body. The method proposed has the potential to become a powerful non-invasive tool for assessment of cutaneous photoaging.

Multimodal imaging of vocal fold scarring in a rabbit model by multiphoton microscopy

NASA Astrophysics Data System (ADS)

Kazarine, Alexei; Bouhabel, Sarah; Douillette, Annie H.; Kost, Karen; Li-Jessen, Nicole Y. K.; Mongeau, Luc; Wiseman, Paul W.

2017-02-01

Vocal fold scarring as a result of injury or disease can lead to voice disorders which can significantly affect the quality of life. During the scarring process, the normally elastic tissue of the vocal fold lamina propria is replaced by a much stiffer collagen-based fibrotic tissue, which impacts the fold's ability to vibrate. Surgical removal of this tissue is often ineffective and can result in further scarring. Injectable biomaterials, a form of tissue engineering, have been proposed as a potential solution to reduce existing scars or prevent scarring altogether. In order to properly evaluate the effectiveness of these new materials, multiphoton microscopy emerges as an effective tool due to its intrinsic multiple label free contrast mechanisms that highlight extracellular matrix elements. In this study, we evaluate the spatial distribution of collagen and elastin fibers in a rabbit model using second harmonic generation (SHG), third harmonic generation (THG) and two photon autofluorescence (TPAF) applied to unlabeled tissue sections. In comparison to traditional methods that rely on histological staining or immunohistochemistry, SHG, THG and TPAF provide a more reliable detection of these native proteins. The evaluation of collagen levels allows us to follow the extent of scarring, while the presence of elastin fibers is thought to be indicative of the level of healing of the injured fold. Using these imaging modalities, we characterize the outcome of injectable biomaterial treatments in order to direct future treatments for tissue engineering.

Three-dimensional simulation of thermal harmonic lasing free electron laser with detuning of the fundamental

DOE Office of Scientific and Technical Information (OSTI.GOV)

Salehi, E.; Maraghechi, B., E-mail: behrouz@aut.ac.ir; School of Particle and Accelerator Physics, Institute for Research in Fundamental Sciences

2016-03-15

Detuning of the fundamental is a way to enhance harmonic generation. By this method, the wiggler is composed of two segments in such a way that the fundamental resonance of the second segment to coincide with the third harmonic of the first segment of the wiggler to generate extreme ultraviolet radiation and x-ray emission. A set of coupled, nonlinear, and first-order differential equations in three dimensions describing the evolution of the electron trajectories and the radiation field with warm beam is solved numerically by CYRUS 3D code in the steady-state for two models (1) seeded free electron laser (FEL) andmore » (2) shot noise on the electron beam (self-amplified spontaneous emission FEL). Thermal effects in the form of longitudinal velocity spread are considered. Three-dimensional simulation describes self-consistently the longitudinal spatial dependence of radiation waists, curvatures, and amplitudes together with the evaluation of the electron beam. The evolutions of the transverse modes are investigated for the fundamental resonance and the third harmonic. Also, the effective modes of the third harmonic are studied. In this paper, we found that detuning of the fundamental with shot noise gives more optimistic result than the seeded FEL.« less

Solid-state laser source of narrowband ultraviolet B light for skin disease care

NASA Astrophysics Data System (ADS)

Tarasov, Aleksandr A.; Chu, Hong

2013-03-01

We report about the development of all-solid-state laser source of narrowband UV-B light for medical applications. The device is based on a gain-switched Ti: Sapphire laser with volume Bragg grating, pumped at 532 nm and operating at 931.8 nm, followed by a third harmonic generator and a fiber optic beam homogenizer. The maximum available pulse energy exceeded 5 mJ at 310.6 nm, with a pulse repetition rates of 50 Hz. The output characteristics satisfy the medical requirements for psoriasis and vitiligo treatment. A new optical scheme for third harmonic generation enhancement at moderate levels of input intensities is proposed and investigated. As a result, 40% harmonic efficiency was obtained, when input pulse power was only 300 kW.

Anomalous behavior in the third harmonic generation z response through dispersion induced shape changes and matching χ(3)

NASA Astrophysics Data System (ADS)

Pillai, Rajesh S.; Brakenhoff, G. J.; Müller, M.

2006-09-01

The third harmonic generation (THG) axial response in the vicinity of an interface formed by two isotropic materials of normal dispersion is typically single peaked, with the maximum intensity at the interface position. Here it is shown experimentally that this THG z response may show anomalous behavior—being double peaked with a dip coinciding with the interface position—when the THG contributions from both materials are of similar magnitude. The observed anomalous behavior is explained, using paraxial Gaussian theory, by considering dispersion induced shape changes in the THG z response.

Electron correlations and pre-collision in the re-collision picture of high harmonic generation

NASA Astrophysics Data System (ADS)

Mašín, Zdeněk; Harvey, Alex G.; Spanner, Michael; Patchkovskii, Serguei; Ivanov, Misha; Smirnova, Olga

2018-07-01

We discuss the seminal three-step model and the re-collision picture in the context of high harmonic generation in molecules. In particular, we stress the importance of multi-electron correlation during the first and the third of the three steps of the process: (1) the strong-field ionization and (3) the recombination. We point out how an accurate account of multi-electron correlations during the third recombination step allows one to gauge the importance of pre-collision: the term coined by Eberly (n.d. private communication) to describe unusual pathways during the first, ionization, step.

A new mode of contrast in biological second harmonic generation microscopy.

PubMed

Green, Nicola H; Delaine-Smith, Robin M; Askew, Hannah J; Byers, Robert; Reilly, Gwendolen C; Matcher, Stephen J

2017-10-17

Enhanced image contrast in biological second harmonic imaging microscopy (SHIM) has previously been reported via quantitative assessments of forward- to epi-generated signal intensity ratio and by polarization analysis. Here we demonstrate a new form of contrast: the material-specific, wavelength-dependence of epi-generated second harmonic generation (SHG) excitation efficiency, and discriminate collagen and myosin by ratiometric epi-generated SHG images at 920 nm and 860 nm. Collagen shows increased SHG intensity at 920 nm, while little difference is detected between the two for myosin; allowing SHIM to characterize different SHG-generating components within a complex biological sample. We propose that momentum-space mapping of the second-order non-linear structure factor is the source of this contrast and develop a model for the forward and epi-generated SHG wavelength-dependence. Our model demonstrates that even very small changes in the assumed material fibrillar structure can produce large changes in the wavelength-dependency of epi-generated SHG. However, in the case of forward SHG, although the same changes impact upon absolute intensity at a given wavelength, they have very little effect on wavelength-dependency beyond the expected monotonic fall. We also propose that this difference between forward and epi-generated SHG provides an explanation for many of the wavelength-dependency discrepancies in the published literature.

Singularity-driven second- and third-harmonic generation at {epsilon}-near-zero crossing points

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vincenti, M. A.; Ceglia, D. de; Ciattoni, A.

We show an alternative path to efficient second- and third-harmonic generation in proximity of the zero crossing points of the dielectric permittivity in conjunction with low absorption. Under these circumstances, any material, either natural or artificial, will show similar degrees of field enhancement followed by strong harmonic generation, without resorting to any resonant mechanism. The results presented in this paper provide a general demonstration of the potential that the zero-crossing-point condition holds for nonlinear optical phenomena. We investigate a generic Lorentz medium and demonstrate that a singularity-driven enhancement of the electric field may be achieved even in extremely thin layersmore » of material. We also discuss the role of nonlinear surface sources in a realistic scenario where a 20-nm layer of CaF{sub 2} is excited at 21 {mu}m, where {epsilon}{approx} 0. Finally, we show similar behavior in an artificial composite material that includes absorbing dyes in the visible range, provide a general tool for the improvement of harmonic generation using the {epsilon}{approx} 0 condition, and illustrate that this singularity-driven enhancement of the field lowers the thresholds for a plethora of nonlinear optical phenomena.« less

Photovoltaic power converter system with a controller configured to actively compensate load harmonics

DOEpatents

de Rooij, Michael Andrew; Steigerwald, Robert Louis; Delgado, Eladio Clemente

2008-12-16

Photovoltaic power converter system including a controller configured to reduce load harmonics is provided. The system comprises a photovoltaic array and an inverter electrically coupled to the array to generate an output current for energizing a load connected to the inverter and to a mains grid supply voltage. The system further comprises a controller including a first circuit coupled to receive a load current to measure a harmonic current in the load current. The controller includes a second circuit to generate a fundamental reference drawn by the load. The controller further includes a third circuit for combining the measured harmonic current and the fundamental reference to generate a command output signal for generating the output current for energizing the load connected to the inverter. The photovoltaic system may be configured to compensate harmonic currents that may be drawn by the load.

Characterization of human carotid atherosclerotic tissues imaged by combining multiple multiphoton microscopy techniques

NASA Astrophysics Data System (ADS)

Baria, E.; Cicchi, R.; Nesi, G.; Massi, D.; Pavone, F. S.

2017-07-01

We combined Second Harmonic Generation, Two-Photon Fluorescence and Fluorescence Lifetime Imaging Microscopy for studying human carotid ex vivo tissue sections affected by atherosclerosis, resulting in the discrimination of different arterial regions within the plaques.

Separation of High Order Harmonics with Fluoride Windows

DOE Office of Scientific and Technical Information (OSTI.GOV)

Allison, Tom; van Tilborg, Jeroen; Wright, Travis

2010-08-02

The lower orders produced in high order harmonic generation can be effciently temporally separated into monochromatic pulses by propagation in a Fluoride window while still preserving their femtosecond pulse duration. We present calculations for MgF2, CaF2, and LiF windows for the third, fifth, and seventh harmonics of 800 nm. We demonstrate the use of this simple and inexpensive technique in a femtosecond pump/probe experiment using the fifth harmonic.

Third harmonic generation of a short pulse laser in a plasma density ripple created by a machining beam

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, C. S.; Tripathi, V. K.

An intense machining laser beam, impinged on a gas jet target, causes space periodic ionization of the gas and heats the electrons. The nonuniform plasma pressure leads to atomic density redistribution. When, after a suitable time delay, a second more intense laser pulse is launched along the periodicity wave vector q-vector, a plasma density ripple n{sub q} is instantly created, leading to resonant third harmonic generation when q=4{omega}{sub p}{sup 2}/(3{omega}c{gamma}{sub 0}), where {omega}{sub p} is the plasma frequency, {omega} is the laser frequency, and {gamma}{sub 0} is the electron Lorentz factor. The third harmonic is produced through the beating ofmore » ponderomotive force induced second harmonic density oscillations and the quiver velocity of electrons at the fundamental. The relativistic mass nonlinearity plays no role in resonant coupling. The energy conversion efficiency scales as the square of plasma density and square of depth of density ripple, and is {approx}0.2% for normalized laser amplitude a{sub o}{approx}1 in a plasma of 1% critical density with 20% density ripple. The theory explains several features of a recent experiment.« less

Harmonic generation with an ultra-strongly coupled cavity polariton

NASA Astrophysics Data System (ADS)

Crescimanno, Michael; Singer, Kenneth; Liu, Bin; McMaster, Michael

2017-04-01

The large dipole density in a new class of glassy organic dyes results in ultrastrong exciton-cavity field coupling leading to polariton splittings of over an eV. We describe the theoretical model and experimental protocol used to understand third harmonic generation (THG) in this system. We quantify the THG enhancement at the polariton branches through its dependence on coupling, cavity-exciton detuning and cavity finesse.

Harmonic generation in metallic, GaAs-filled nanocavities in the enhanced transmission regime at visible and UV wavelengths.

PubMed

Vincenti, M A; de Ceglia, D; Roppo, V; Scalora, M

2011-01-31

We have conducted a theoretical study of harmonic generation from a silver grating having slits filled with GaAs. By working in the enhanced transmission regime, and by exploiting phase-locking between the pump and its harmonics, we guarantee strong field localization and enhanced harmonic generation under conditions of high absorption at visible and UV wavelengths. Silver is treated using the hydrodynamic model, which includes Coulomb and Lorentz forces, convection, electron gas pressure, plus bulk χ(3) contributions. For GaAs we use nonlinear Lorentz oscillators, with characteristic χ(2) and χ(3) and nonlinear sources that arise from symmetry breaking and Lorentz forces. We find that: (i) electron pressure in the metal contributes to linear and nonlinear processes by shifting/reshaping the band structure; (ii) TE- and TM-polarized harmonics can be generated efficiently; (iii) the χ(2) tensor of GaAs couples TE- and TM-polarized harmonics that create phase-locked pump photons having polarization orthogonal compared to incident pump photons; (iv) Fabry-Perot resonances yield more efficient harmonic generation compared to plasmonic transmission peaks, where most of the light propagates along external metal surfaces with little penetration inside its volume. We predict conversion efficiencies that range from 10(-6) for second harmonic generation to 10(-3) for the third harmonic signal, when pump power is 2 GW/cm2.

Spontaneous Polarization in Bio-organic Materials Studied by Scanning Pyroelectric Microscopy (SPEM) and Second Harmonic Generation Microscopy (SHGM)

NASA Astrophysics Data System (ADS)

Putzeys, T.; Wübbenhorst, M.; van der Veen, M. A.

2015-06-01

Bio-organic materials such as bones, teeth, and tendon generally show nonlinear optical (Masters and So in Handbook of Biomedical Nonlinear Optical Microscopy, 2008), pyro- and piezoelectric (Fukada and Yasuda in J Phys Soc Jpn 12:1158, 1957) properties, implying a permanent polarization, the presence of which can be rationalized by describing the growth of the sample and the creation of a polar axis according to Markov's theory of stochastic processes (Hulliger in Biophys J 84:3501, 2003; Batagiannis et al. in Curr Opin Solid State Mater Sci 17:107, 2010). Two proven, versatile techniques for probing spontaneous polarization distributions in solids are scanning pyroelectric microscopy (SPEM) and second harmonic generation microscopy (SHGM). The combination of pyroelectric scanning with SHG-microscopy in a single experimental setup leading to complementary pyroelectric and nonlinear optical data is demonstrated, providing us with a more complete image of the polarization in organic materials. Crystals consisting of a known polar and hyperpolarizable material, CNS (4-chloro-4-nitrostilbene) are used as a reference sample, to verify the functionality of the setup, with both SPEM and SHGM images revealing the same polarization domain information. In contrast, feline and human nails exhibit a pyroelectric response, but a second harmonic response is absent for both keratin containing materials, implying that there may be symmetry-allowed SHG, but with very inefficient second harmonophores. This new approach to polarity detection provides additional information on the polar and hyperpolar nature in a variety of (bio) materials.

Molecular Orientation in Dry and Hydrated Cellulose Fibers: A Coherent Anti-Stokes Raman Scattering Microscopy Study

PubMed Central

Zimmerley, Maxwell; Younger, Rebecca; Valenton, Tiffany; Oertel, David C.; Ward, Jimmie L.; Potma, Eric O.

2012-01-01

Coherent anti-Stokes Raman scattering (CARS) microscopy is combined with spontaneous Raman scattering microspectroscopy and second harmonic generation (SHG) microscopy to interrogate the molecular alignment in dry and hydrated cellulose fibers. Two types of cellulose were investigated: natural cellulose I in cotton fibers and regenerated cellulose II in rayon fibers. On the basis of the orientation of the methylene symmetric stretching vibration, the molecular alignment of cellulose microfibrils is found to be conserved on the micrometer scale. Whereas the molecular orientation in cotton shows modest variability along the fiber, the alignment of the cellulose units in rayon is highly consistent throughout the fiber. The ordered alignment is retained upon fiber hydration. Upon hydration of the cellulose fibers, an anisotropic electronic contribution is observed, which indicates an ordered incorporation of water molecules into the fiber structure. The third-order and second-order electronic polarizability of cellulose I are directed along the axis of the polyglucan chain. No second-order optical response is observed in cellulose II, supporting the antiparallel arrangement of the polyglucan chains in regenerated cellulose. PMID:20684644

Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure

PubMed Central

Chen, Xiyi; Nadiarynkh, Oleg; Plotnikov, Sergey; Campagnola, Paul J

2013-01-01

Second-harmonic generation (SHG) microscopy has emerged as a powerful modality for imaging fibrillar collagen in a diverse range of tissues. Because of its underlying physical origin, it is highly sensitive to the collagen fibril/fiber structure, and, importantly, to changes that occur in diseases such as cancer, fibrosis and connective tissue disorders. We discuss how SHG can be used to obtain more structural information on the assembly of collagen in tissues than is possible by other microscopy techniques. We first provide an overview of the state of the art and the physical background of SHG microscopy, and then describe the optical modifications that need to be made to a laser-scanning microscope to enable the measurements. Crucial aspects for biomedical applications are the capabilities and limitations of the different experimental configurations. We estimate that the setup and calibration of the SHG instrument from its component parts will require 2–4 weeks, depending on the level of the user’s experience. PMID:22402635

Optical control of recovery speed of photoinduced third-harmonic generation in azo-copolymer thin films

NASA Astrophysics Data System (ADS)

Lin, Jian Hung; Lai, Ngoc Diep; Hsu, Chia Chen

2006-03-01

Recovery speed of photoinduced third-harmonic (TH) generation in azo-copolymer thin films can be controlled by a nanosecond laser excitation. When the excitation is tuned on, the TH signal decreases because of angular hole burning and angular redistribution effects. After turning off the excitation, the TH signal can recover to its original level either within 1min (high intensity excitation) or longer than several days (low intensity excitation). The fast recovery of the TH signal is attributed to the increase of temperature in the sample that causes molecules to more easily reorient and return to the original trans form.

Third-harmonic generation of a laser-driven quantum dot with impurity

NASA Astrophysics Data System (ADS)

Sakiroglu, S.; Kilic, D. Gul; Yesilgul, U.; Ungan, F.; Kasapoglu, E.; Sari, H.; Sokmen, I.

2018-06-01

The third-harmonic generation (THG) coefficient for a laser-driven quantum dot with an on-center Gaussian impurity under static magnetic field is theoretically investigated. Laser field effect is treated within the high-frequency Floquet approach and the analytical expression of the THG coefficient is deduced from the compact density-matrix approach. The numerical results demonstrate that the application of intense laser field causes substantial changes on the behavior of THG. In addition the position and magnitude of the resonant peak of THG coefficient is significantly affected by the magnetic field, quantum dot size and the characteristic parameters of the impurity potential.

Collagen analysis by second-harmonic generation microscopy predicts outcome of luminal breast cancer.

PubMed

Natal, Rodrigo A; Vassallo, José; Paiva, Geisilene R; Pelegati, Vitor B; Barbosa, Guilherme O; Mendonça, Guilherme R; Bondarik, Caroline; Derchain, Sophie F; Carvalho, Hernandes F; Lima, Carmen S; Cesar, Carlos L; Sarian, Luís Otávio

2018-04-01

Second-harmonic generation microscopy represents an important tool to evaluate extracellular matrix collagen structure, which undergoes changes during cancer progression. Thus, it is potentially relevant to assess breast cancer development. We propose the use of second-harmonic generation images of tumor stroma selected on hematoxylin and eosin-stained slides to evaluate the prognostic value of collagen fibers analyses in peri and intratumoral areas in patients diagnosed with invasive ductal breast carcinoma. Quantitative analyses of collagen parameters were performed using ImageJ software. These parameters presented significantly higher values in peri than in intratumoral areas. Higher intratumoral collagen uniformity was associated with high pathological stages and with the presence of axillary lymph node metastasis. In patients with immunohistochemistry-based luminal subtype, higher intratumoral collagen uniformity and quantity were independently associated with poorer relapse-free and overall survival, respectively. A multivariate response recursive partitioning model determined 12.857 and 11.894 as the best cut-offs for intratumoral collagen quantity and uniformity, respectively. These values have shown high sensitivity and specificity to differentiate distinct outcomes. Values of intratumoral collagen quantity and uniformity exceeding the cut-offs were strongly associated with poorer relapse-free and overall survival. Our findings support a promising prognostic value of quantitative evaluation of intratumoral collagen by second-harmonic generation imaging mainly in the luminal subtype breast cancer.

Characterization of second and third order optical nonlinearities of ZnO sputtered films

NASA Astrophysics Data System (ADS)

Larciprete, M. C.; Haertle, D.; Belardini, A.; Bertolotti, M.; Sarto, F.; Günter, P.

2006-03-01

We measured the second and third order optical nonlinearity of zinc oxide, grown on glass substrates by the ion beam sputtering technique. Second and third harmonic generation measurements were performed by means of the rotational Maker fringes technique for different polarization configurations, thus allowing the determination of all non-zero components of the second order susceptibility at three different fundamental beam wavelengths, i.e., 1064 nm, 1542 nm and 1907 nm. The dispersion of the nonlinear optical coefficients has been evaluated, while the nonlinear optical coefficients were found to range between 0.9 pm/V and 0.16 pm/V for d33, 0.53 pm/V and 0.08 pm/V for |d15|, 0.31 and 0.08 pm/V for |d31|, with increasing wavelength. Finally, one third order susceptibility, χijkl (3), has been determined by third harmonic generation measurements at a fundamental wavelength λ=1907 nm and a value for χ3333 (3) of 185×10-20 m2/V2 has been found.

Ex-vivo multiphoton analysis of rabbit corneal wound healing following photorefractive keratectomy

NASA Astrophysics Data System (ADS)

Wang, Tsung-Jen; Lo, Wen; Dong, Chen-Yuan; Hu, Fung-Rong

2008-02-01

The aim of this study is to assess the application of multiphoton autofluorescence and second harmonic generation (SHG) microscopy for investigating corneal wound healing after high myopic (-10.0D) photorefractive keratectomy (PRK) procedures on the rabbit eyes. The effect of PRK on the morphology and distribution of keratocytes were investigated using multiphoton excited autofluorescence imaging, while the effect of PRK on the arrangement of collagen fibers was monitored by second-harmonic generation imaging. Without histological processing, multiphoton microscopy is able to characterize corneal damage and wound healing from PRK. Our results show that this technique has potential application in the clinical evaluation of corneal damage due to refractive surgery, and may be used to study the unwanted side effects of these procedures.

On energy harvesting for augmented tags

NASA Astrophysics Data System (ADS)

Allane, Dahmane; Duroc, Yvan; Andia Vera, Gianfranco; Touhami, Rachida; Tedjini, Smail

2017-02-01

In this paper, the harmonic signals generated by UHF RFID chips, usually considered as spurious effects and unused, are exploited. Indeed, the harmonic signals are harvested to feed a supplementary circuitry associated with a passive RFID tag. Two approaches are presented and compared. In the first one, the third-harmonic signal is combined with an external 2.45-GHz Wi-Fi signal. The integration is done in such a way that the composite signal boosts the conversion efficiency of the energy harvester. In the second approach, the third-harmonic signal is used as the only source of a harvester that energizes a commercial temperature sensor associated with the tag. The design procedures of the two "augmented-tag" approaches are presented. The performance of each system is simulated with ADS software, and using Harmonic Balance tool (HB), the results obtained in simulation and measurements are compared also. xml:lang="fr"

Ophthalmic imaging using multiphoton microscopy

NASA Astrophysics Data System (ADS)

Teng, Shu-Wen; Peng, Ju-Li; Lin, Huei-Hsing; Wu, Hai-Yin; Lo, Wen; Sun, Yen; Lin, Wei-Chou; Lin, Sung-Jan; Jee, Shiou-Hwa; Tan, Hsin-Yuan; Dong, Chen-Yuan

2005-04-01

This purpose of this study is to demonstrate the feasibility of using multiphoton microscopy in ophthalmologic imaging. Without the introduction of extrinsic fluorescence molecules, multiphoton induced autofluorescence and second harmonic generation signals can be used to obtain useful structural information of normal and diseased corneas. Our work can potentially lead to the in vivo application of multiphoton microscopy in investigating corneal physiology and pathologies.

Three-dimensional structural imaging of starch granules by second-harmonic generation circular dichroism.

PubMed

Zhuo, G-Y; Lee, H; Hsu, K-J; Huttunen, M J; Kauranen, M; Lin, Y-Y; Chu, S-W

2014-03-01

Chirality is one of the most fundamental and essential structural properties of biological molecules. Many important biological molecules including amino acids and polysaccharides are intrinsically chiral. Conventionally, chiral species can be distinguished by interaction with circularly polarized light, and circular dichroism is one of the best-known approaches for chirality detection. As a linear optical process, circular dichroism suffers from very low signal contrast and lack of spatial resolution in the axial direction. It has been demonstrated that by incorporating nonlinear interaction with circularly polarized excitation, second-harmonic generation circular dichroism can provide much higher signal contrast. However, previous circular dichroism and second-harmonic generation circular dichroism studies are mostly limited to probe chiralities at surfaces and interfaces. It is known that second-harmonic generation, as a second-order nonlinear optical effect, provides excellent optical sectioning capability when combined with a laser-scanning microscope. In this work, we combine the axial resolving power of second-harmonic generation and chiral sensitivity of second-harmonic generation circular dichroism to realize three-dimensional chiral detection in biological tissues. Within the point spread function of a tight focus, second-harmonic generation circular dichroism could arise from the macroscopic supramolecular packing as well as the microscopic intramolecular chirality, so our aim is to clarify the origins of second-harmonic generation circular dichroism response in complicated three-dimensional biological systems. The sample we use is starch granules whose second-harmonic generation-active molecules are amylopectin with both microscopic chirality due to its helical structure and macroscopic chirality due to its crystallized packing. We found that in a starch granule, the second-harmonic generation for right-handed circularly polarized excitation is significantly different from second-harmonic generation for left-handed one, offering excellent second-harmonic generation circular dichroism contrast that approaches 100%. In addition, three-dimensional visualization of second-harmonic generation circular dichroism distribution with sub-micrometer spatial resolution is realized. We observed second-harmonic generation circular dichroism sign change across the starch granules, and the result suggests that in thick biological tissue, second-harmonic generation circular dichroism arises from macroscopic molecular packing. Our result provides a new method to visualize the organization of three-dimensional structures of starch granules. The second-harmonic generation circular dichroism imaging method expands the horizon of nonlinear chiroptical studies from simplified surface/solution environments to complicated biological tissues. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

Third-harmonic entanglement and Einstein-Podolsky-Rosen steering over a frequency range of more than an octave

NASA Astrophysics Data System (ADS)

Olsen, M. K.

2018-03-01

The development of quantum technologies which use quantum states of the light field interacting with other systems creates a demand for such states over wide frequency ranges. In this work we compare the bipartite entanglement and Einstein-Podolsky-Rosen (EPR) -steering properties of the two different parametric schemes which produce third-harmonic optical fields from an input field at the fundamental frequency. The first scheme uses second harmonic cascaded with sum-frequency generation, while the second uses triply degenerate four- wave mixing, also known as direct third-harmonic generation. We find that both schemes produce continuous-variable bipartite entanglement and EPR steering over a frequency range which has previously been unobtainable. The direct scheme produces a greater degree of EPR steering, while the cascaded scheme allows for greater flexibility in having three available bipartitions, thus allowing for greater flexibility in the tailoring of light matter interfaces. There are also parameter regimes in both for which classical mean-field analyses fail to predict the mean-field solutions. Both schemes may be very useful for applications in quantum communication and computation networks, as well as providing for quantum interfaces between a wider range of light and atomic ensembles than is presently practicable.

Self-referenced axial chromatic dispersion measurement in multiphoton microscopy through 2-color THG imaging.

PubMed

Du, Yu; Zhuang, Ziwei; He, Jiexing; Liu, Hongji; Qiu, Ping; Wang, Ke

2018-05-16

With tunable excitation light, multiphoton microscopy (MPM) is widely used for imaging biological structures at subcellular resolution. Axial chromatic dispersion, present in virtually every transmissive optical system including the multiphoton microscope, leads to focal (and the resultant image) plane separation. Here we demonstrate experimentally a technique to measure the axial chromatic dispersion in a multiphoton microscope, using simultaneous 2-color third-harmonic generation (THG) imaging excited by a 2-color soliton source with tunable wavelength separation. Our technique is self-referenced, eliminating potential measurement error when 1-color tunable excitation light is used which necessitates reciprocating motion of the mechanical translation stage. Using this technique, we demonstrate measured axial chromatic dispersion with 2 different objective lenses in a multiphoton microscope. Further measurement in a biological sample also indicates that this axial chromatic dispersion, in combination with 2-color imaging, may open up opportunity for simultaneous imaging of two different axial planes. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Generation of higher odd harmonics in a defective photonic crystal

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ramanujam, N. R., E-mail: wilsonpra@yahoo.co.in; Wilson, K. S. Joseph

2015-06-24

A photonic crystal (AB){sup 2}(DB)(AB){sup 2} with high refractive index medium as silicon and low refractive medium as air is considered. Using the transfer matrix method, the transmission properties as a function of wavelength with photonic band gaps has been obtained. We are able to demonstrate the generation of third, fifth, seventh and ninth harmonics in the present work. We show that if the air medium is removed in the defect, the defect modes are generated but not harmonics. It can be designed to have a frequency conversion, and have a potential for becoming the basis for the next generationmore » of optical devices.« less

Improvement and formation of UV-induced damage on LBO crystal surface during long-term high-power third-harmonic generation.

PubMed

Hong, Hailong; Liu, Qiang; Huang, Lei; Gong, Mali

2013-03-25

We demonstrate the improvement and formation of UV-induced damage on LBO crystal output surface during long-term (130 h) high-power (20 W) high-repetition-rate (80 kHz) third-harmonic generation. The output surface was super-polished (RMS surface roughness <0.6 nm) to sub-nanometer scale super smooth roughness. The surface lifetime has been improved more than 20-fold compared with the as-polished ones (RMS surface roughness 4.0~8.0 nm). The damage could be attributed to the consequence of thermal effects resulted from impurity absorptions. Simultaneously, it was verified that the impurities originated in part from the UV-induced deposition.

Detecting a pronounced delocalized state in third-harmonic generation phenomenon; a quantum chaos approach

NASA Astrophysics Data System (ADS)

Behnia, S.; Ziaei, J.; Khodavirdizadeh, M.

2018-06-01

Nonlinear optics (NLO) deserves special attention in new optical devices, making it possible to generate coherent light more efficiently. Among the various NLO phenomena the third-harmonic generation (THG) is at the core of the effective operating mechanism of broadband wavelength conversion, in all-optical devices. Here, we aim to understand how the third-order susceptibility and the electric field may be effectively effect on the localization properties of the light in the THG process when included in a two-mode cavity coherently perturbed by a classical field. We address a stable-unstable transition due to the combination effect of the aforementioned factors. We report a reliable evidence confirming the appearance of chaos in THG under suitable conditions. By tracing the signatures of adjacent-spectral-spacing-ratio (ASSR) distribution and participation ratio, we also find a critical point (ɛc ,κc) =(3 . 1 , 0 . 35) for which a pronounced delocalized response is seen. This study may have profound findings for practical devices, and ushers in new opportunities for practical exploitation of the electric field and the third-order susceptibility effect in nonlinear optical devices.

Coherent nonlinear optical response of single-layer black phosphorus: third-harmonic generation

NASA Astrophysics Data System (ADS)

Margulis, Vladimir A.; Muryumin, Evgeny E.; Gaiduk, Evgeny A.

2017-10-01

We theoretically calculate the nonlinear optical (NLO) response of phosphorene (a black phosphorus monolayer) to a normally incident and linearly polarized coherent laser radiation of frequency ω, resulting in the generation of radiation at frequency 3ω. We derive explicit analytic expressions for four independent nonvanishing elements of the third-order NLO susceptibility tensor, describing the third-harmonic generation (THG) from phosphorene. The final formulas are numerically evaluated for typical values of the system's parameters to explore how the efficiency of the THG varies with both the frequency and the polarization direction of the incident radiation. The results obtained show a resonant enhancement of the THG efficiency when the pump photon energy ℏω approaches a value of one third of the bandgap energy Eg (≈1.5 eV) of phosphorene. It is also shown that the THG efficiency exhibits a specific polarization dependence, allowing the THG to be used for determining the orientation of phosphorene's crystallographic axes. Our findings highlight the material's potential for practical application in nanoscale photonic devices such as frequency convertors operating in the near-infrared spectral range.

Thermally and electrically controllable multiple high harmonics generation by harmonically driven quasi-two-dimensional electron gas

NASA Astrophysics Data System (ADS)

Maglevanny, I. I.; Smolar, V. A.; Karyakina, T. I.

2018-06-01

In this paper, we consider the activation processes in nonlinear meta-stable system based on a lateral (quasi-two-dimensional) superlattice and study the dynamics of such a system externally driven by a harmonic force. The internal control parameters are the longitudinal applied electric field and the sample temperature. The spontaneous transverse electric field is considered as an order parameter. The forced violations of order parameter are considered as a response of a system to periodic driving. We investigate the cooperative effects of self-organization and high harmonic forcing from the viewpoint of catastrophe theory and show the possibility of generation of third and higher odd harmonics in output signal that lead to distortion of its wave front. A higher harmonics detection strategy is further proposed and explained in detail by exploring the influences of system parameters on the response output of the system that are discussed through numerical simulations.

Electromagnetic probes of molecular motors in the electron transport chains of mitochondria and chloroplasts

NASA Astrophysics Data System (ADS)

Miller, J. H., Jr.; Nawarathna, D.; Vajrala, V.; Gardner, J.; Widger, W. R.

2005-12-01

We report on measurements of harmonics generated by whole cells, mitochondria, and chloroplasts in response to applied sinusoidal electric fields. The frequency- and amplitude-dependence of the induced harmonics exhibit features that correlate with physiological processes. Budding yeast (S. cerevisiae) cells produce numerous harmonics, the amplitudes of which depend strongly on frequency. When the second or third harmonic amplitude is plotted vs. applied frequency, we observe two peaks, around 3 kHz and 12 kHz, which are suppressed by respiratory inhibitors. We observe similar peaks when measuring the harmonic response of B. indicas, a relative of the mitochondrial ancestor. In uncoupled mitochondria, in which most of the electron transport chain is active but the ATP-synthase molecular turbine is inactive, only one (lower frequency) of the two peaks is present. Finally, we find that harmonics generated by chloroplasts depend dramatically on incident light, and vanish in the absence of light.

Involvement of small carbon clusters in the enhancement of high-order harmonic generation of ultrashort pulses in the plasmas produced during ablation of carbon-contained nanoparticles

NASA Astrophysics Data System (ADS)

Ganeev, R. A.

2017-09-01

Various carbon-based nanoparticles ablated at the conditions suitable for efficient harmonic generation during propagation of ultrashort pulses through the laser-produced plasmas were studied. The transmission electron microscopy of ablated debris and the time-of-flight mass-spectroscopy studies of plasmas are presented. The conditions of laser ablation of the carbon-contained nanoparticles (fullerenes, graphene, carbon nanotubes, carbon nanofibers, and diamond nanoparticles) were varied to define the impeding processes restricting the harmonic yield from such laser-produced plasmas. These studies show that the enhancement of harmonics during ablation of nanoparticle targets was related with the appearance of small carbon clusters at the moment of propagation of the ultrashort laser pulses though such plasmas.

Quantitative imaging of fibrotic and morphological changes in liver of non-alcoholic steatohepatitis (NASH) model mice by second harmonic generation (SHG) and auto-fluorescence (AF) imaging using two-photon excitation microscopy (TPEM).

PubMed

Yamamoto, Shin; Oshima, Yusuke; Saitou, Takashi; Watanabe, Takao; Miyake, Teruki; Yoshida, Osamu; Tokumoto, Yoshio; Abe, Masanori; Matsuura, Bunzo; Hiasa, Yoichi; Imamura, Takeshi

2016-12-01

Non-alcoholic steatohepatitis (NASH) is a common liver disorder caused by fatty liver. Because NASH is associated with fibrotic and morphological changes in liver tissue, a direct imaging technique is required for accurate staging of liver tissue. For this purpose, in this study we took advantage of two label-free optical imaging techniques, second harmonic generation (SHG) and auto-fluorescence (AF), using two-photon excitation microscopy (TPEM). Three-dimensional ex vivo imaging of tissues from NASH model mice, followed by image processing, revealed that SHG and AF are sufficient to quantitatively characterize the hepatic capsule at an early stage and parenchymal morphologies associated with liver disease progression, respectively.

Kinetic Modeling of Accelerated Stability Testing Enabled by Second Harmonic Generation Microscopy.

PubMed

Song, Zhengtian; Sarkar, Sreya; Vogt, Andrew D; Danzer, Gerald D; Smith, Casey J; Gualtieri, Ellen J; Simpson, Garth J

2018-04-03

The low limits of detection afforded by second harmonic generation (SHG) microscopy coupled with image analysis algorithms enabled quantitative modeling of the temperature-dependent crystallization of active pharmaceutical ingredients (APIs) within amorphous solid dispersions (ASDs). ASDs, in which an API is maintained in an amorphous state within a polymer matrix, are finding increasing use to address solubility limitations of small-molecule APIs. Extensive stability testing is typically performed for ASD characterization, the time frame for which is often dictated by the earliest detectable onset of crystal formation. Here a study of accelerated stability testing on ritonavir, a human immunodeficiency virus (HIV) protease inhibitor, has been conducted. Under the condition for accelerated stability testing at 50 °C/75%RH and 40 °C/75%RH, ritonavir crystallization kinetics from amorphous solid dispersions were monitored by SHG microscopy. SHG microscopy coupled by image analysis yielded limits of detection for ritonavir crystals as low as 10 ppm, which is about 2 orders of magnitude lower than other methods currently available for crystallinity detection in ASDs. The four decade dynamic range of SHG microscopy enabled quantitative modeling with an established (JMAK) kinetic model. From the SHG images, nucleation and crystal growth rates were independently determined.

Ultrafast Optical Modulation of Second- and Third-Harmonic Generation from Cut-Disk-Based Metasurfaces

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sartorello, Giovanni; Olivier, Nicolas; Zhang, Jingjing

2016-08-17

We design and fabricate a metasurface composed of gold cut-disk resonators that exhibits a strong coherent nonlinear response. We experimentally demonstrate all-optical modulation of both second- and third-harmonic signals on a subpicosecond time scale. Pump probe experiments and numerical models show that the observed effects are due to the ultrafast response of the electronic excitations in the metal under external illumination. These effects pave the way for the development of novel active nonlinear metasurfaces with controllable and switchable coherent nonlinear response.

Modulating optical rectification, second and third harmonic generation of doped quantum dots: Interplay between hydrostatic pressure, temperature and noise

NASA Astrophysics Data System (ADS)

Ganguly, Jayanta; Saha, Surajit; Bera, Aindrila; Ghosh, Manas

2016-10-01

We examine the profiles of optical rectification (OR), second harmonic generation (SHG) and third harmonic generation (THG) of impurity doped QDs under the combined influence of hydrostatic pressure (HP) and temperature (T) in presence and absence of Gaussian white noise. Noise has been incorporated to the system additively and multiplicatively. In order to study the above nonlinear optical (NLO) properties the doped dot has been subjected to a polarized monochromatic electromagnetic field. Effect of application of noise is nicely reflected through alteration of peak shift (blue/red) and variation of peak height (increase/decrease) of above NLO properties as temperature and pressure are varied. All such changes again sensitively depends on mode of application (additive/multiplicative) of noise. The remarkable influence of interplay between noise strength and its mode of application on the said profiles has also been addressed. The findings illuminate fascinating role played by noise in tuning above NLO properties of doped QD system under the active presence of both hydrostatic pressure and temperature.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Murase, Kenya, E-mail: murase@sahs.med.osaka-u.ac.jp; Song, Ruixiao; Hiratsuka, Samu

We investigated the feasibility of visualizing blood coagulation using a system for magnetic particle imaging (MPI). A magnetic field-free line is generated using two opposing neodymium magnets and transverse images are reconstructed from the third-harmonic signals received by a gradiometer coil, using the maximum likelihood-expectation maximization algorithm. Our MPI system was used to image the blood coagulation induced by adding CaCl{sub 2} to whole sheep blood mixed with magnetic nanoparticles (MNPs). The “MPI value” was defined as the pixel value of the transverse image reconstructed from the third-harmonic signals. MPI values were significantly smaller for coagulated blood samples than thosemore » without coagulation. We confirmed the rationale of these results by calculating the third-harmonic signals for the measured viscosities of samples, with an assumption that the magnetization and particle size distribution of MNPs obey the Langevin equation and log-normal distribution, respectively. We concluded that MPI can be useful for visualizing blood coagulation.« less

A high repetition rate multiwavelength polarized solid state laser source for long range lidar applications

NASA Astrophysics Data System (ADS)

Sudheer, S. K.; Pillai, V. P. Mahadevan; Nayar, V. U.

2006-12-01

Advances in Laser Technology and nonlinear Optical techniques can be effectively utilized for LIDAR applications in space and atmospheric sciences to achieve better flexibility and control of the available optical power. Using such devices, one can achieve highly accurate and resolved, measurement of the distribution for atmospheric scattering layers. In the present investigation a diode double end pumped high repetition rate, multi wavelength Nd:YAG laser is designed, fabricated and various laser beam parameters have been characterized for LIDAR applications. Nonlinear optical techniques have been employed to generate higher harmonics like 532nm, 355nm and 266nm for various spectral studies. The experimental setup mainly consists of two Fiber coupled pump laser diodes (Model FAP- 81-30C-800B, Coherent Inc, USA) with a maximum output power of 30Watt at a wavelength of 807-810nm at 30°C set temperature. A second harmonic LBO crystal cut for critical phase matching placed within the laser resonator is provided for converting a fraction of the fundamental beam to a second harmonic beam. A type II frequency tripling LBO nonlinear crystal (cut for critical phase matching) is also located inside the laser resonator. The third harmonic beam and the unconverted fundamental beam are then directed across a type I fourth harmonic LBO crystal cut for critical phase matching where a portion of the fundamental beam and a portion of the third harmonic beam are converted to a fourth harmonic frequency when both fundamental and third harmonic beams propagate through the frequency quadrupling crystal. The resulting beams which are the fundamental (1064nm), second harmonic (532nm), third harmonic (355nm) and fourth harmonic (266nm) are then directed to a fourth harmonic separator in which the fourth harmonic beam is separated from the fundamental beam. A maximum average power of 12W at 1064nm, 8W at 532nm, 5W at 355nm and 3W at 266nm have been measured at a repetition rate of 10KHz. A minimum pulse width of 25ns have been observed.

Visualization of collagen regeneration in mouse dorsal skin using second harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Luo, T.; Chen, J. X.; Zhuo, S. M.; Lu, K. C.; Jiang, X. S.; Liu, Q. G.

2009-03-01

The purpose of this study is to highlight a clearer understanding of the process of collagen regeneration during wound healing. By means of second harmonic generation (SHG) microscopy, the changes of collagen arrangement at the wound margin were analyzed at 0, 3, 5, 7, 11 and 13 days post injury. The degree of collagen disorders associated with the healing process was quantitatively obtained using the aspect ratio of polar plot image of collagen azimuthal angles and the healing status of collagen could be estimated by arithmetical mean deviation ( Ra) of the collagen SHG images. Our results suggest that SHG microscopy has potential advances in the collagen studies during wound healing and the arrangement of collagen fibers gradually transformed from disorder to order so as to contract the wound. It is capable of promoting clinical application of the noninvasive imaging tool and the analysis methods of collagen disorder as an effective scar management for prevention and treatment about aberrant healing.

Imaging articular cartilage using second harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Mansfield, Jessica C.; Winlove, C. Peter; Knapp, Karen; Matcher, Stephen J.

2006-02-01

Sub cellular resolution images of equine articular cartilage have been obtained using both second harmonic generation microscopy (SHGM) and two-photon fluorescence microscopy (TPFM). The SHGM images clearly map the distribution of the collagen II fibers within the extracellular matrix while the TPFM images show the distribution of endogenous two-photon fluorophores in both the cells and the extracellular matrix, highlighting especially the pericellular matrix and bright 2-3μm diameter features within the cells. To investigate the source of TPF in the extracellular matrix experiments have been carried out to see if it may originate from the proteoglycans. Pure solutions of the following proteoglycans hyaluronan, chondroitin sulfate and aggrecan have been imaged, only the aggrecan produced any TPF and here the intensity was not great enough to account for the TPF in the extracellular matrix. Also cartilage samples were subjected to a process to remove proteoglycans and cellular components. After this process the TPF from the samples had decreased by a factor of two, with respect to the SHG intensity.

Atom-Dependent Edge-Enhanced Second-Harmonic Generation on MoS2 Monolayers.

PubMed

Lin, Kuang-I; Ho, Yen-Hung; Liu, Shu-Bai; Ciou, Jian-Jhih; Huang, Bo-Ting; Chen, Christopher; Chang, Han-Ching; Tu, Chien-Liang; Chen, Chang-Hsiao

2018-02-14

Edge morphology and lattice orientation of single-crystal molybdenum disulfide (MoS 2 ) monolayers, a transition metal dichalcogenide (TMD), possessing a triangular shape with different edges grown by chemical vapor deposition are characterized by atomic force microscopy and transmission electron microscopy. Multiphoton laser scanning microscopy is utilized to study one-dimensional atomic edges of MoS 2 monolayers with localized midgap electronic states, which result in greatly enhanced optical second-harmonic generation (SHG). Microscopic S-zigzag edge and S-Mo Klein edge (bare Mo atoms protruding from a S-zigzag edge) terminations and the edge-atom dependent resonance energies can therefore be deduced based on SHG images. Theoretical calculations based on density functional theory clearly explain the lower energy of the S-zigzag edge states compared to the corresponding S-Mo Klein edge states. Characterization of the atomic-scale variation of edge-enhanced SHG is a step forward in this full-optical and high-yield technique of atomic-layer TMDs.

Cascading second-order nonlinear processes in a lithium niobate-on-insulator microdisk.

PubMed

Liu, Shijie; Zheng, Yuanlin; Chen, Xianfeng

2017-09-15

Whispering-gallery-mode (WGM) microcavities are very important in both fundamental science and practical applications, among which on-chip second-order nonlinear microresonators play an important role in integrated photonic functionalities. Here we demonstrate resonant second-harmonic generation (SHG) and cascaded third-harmonic generation (THG) in a lithium niobate-on-insulator (LNOI) microdisk resonator. Efficient SHG in the visible range was obtained with only several mW input powers at telecom wavelengths. THG was also observed through a cascading process, which reveals simultaneous phase matching and strong mode coupling in the resonator. Cascading of second-order nonlinear processes gives rise to an effectively large third-order nonlinearity, which makes on-chip second-order nonlinear microresonators a promising frequency converter for integrated nonlinear photonics.

Miniaturized video-rate epi-third-harmonic-generation fiber-microscope.

PubMed

Chia, Shih-Hsuan; Yu, Che-Hang; Lin, Chih-Han; Cheng, Nai-Chia; Liu, Tzu-Ming; Chan, Ming-Che; Chen, I-Hsiu; Sun, Chi-Kuang

2010-08-02

With a micro-electro-mechanical system (MEMS) mirror, we successfully developed a miniaturized epi-third-harmonic-generation (epi-THG) fiber-microscope with a video frame rate (31 Hz), which was designed for in vivo optical biopsy of human skin. With a large-mode-area (LMA) photonic crystal fiber (PCF) and a regular microscopic objective, the nonlinear distortion of the ultrafast pulses delivery could be much reduced while still achieving a 0.4 microm lateral resolution for epi-THG signals. In vivo real time virtual biopsy of the Asian skin with a video rate (31 Hz) and a sub-micron resolution was obtained. The result indicates that this miniaturized system was compact enough for the least invasive hand-held clinical use.

Dietary adaptions in the ultrastructure of dinosaur dentine.

PubMed

Brink, Kirstin S; Chen, Yu-Cheng; Wu, Ya-Na; Liu, Wei-Min; Shieh, Dar-Bin; Huang, Timothy D; Sun, Chi-Kuang; Reisz, Robert R

2016-12-01

Teeth are key to understanding the feeding ecology of both extant and extinct vertebrates. Recent studies have highlighted the previously unrecognized complexity of dinosaur dentitions and how specific tooth tissues and tooth shapes differ between taxa with different diets. However, it is unknown how the ultrastructure of these tooth tissues contributes to the differences in feeding style between taxa. In this study, we use third harmonic generation microscopy and scanning electron microscopy to examine the ultrastructure of the dentine in herbivorous and carnivorous dinosaurs to understand how the structure of this tissue contributes to the overall utility of the tooth. Morphometric analyses of dentinal tubule diameter, density and branching rates reveal a strong signal for dietary preferences, with herbivorous saurischian and ornithischian dinosaurs consistently having higher dentinal tubule density than their carnivorous relatives. We hypothesize that this relates to the hardness of the dentine, where herbivorous taxa have dentine that is more resistant to breakage and wear at the dentine-enamel junction than carnivorous taxa. This study advocates the detailed study of dentine and the use of advanced microscopy techniques to understand the evolution of dentition and feeding ecology in extinct vertebrates. © 2016 The Author(s).

Second-harmonic generation from a positive-negative index material heterostructure.

PubMed

Mattiucci, Nadia; D'Aguanno, Giuseppe; Bloemer, Mark J; Scalora, Michael

2005-12-01

Resonant cavities have been widely used in the past to enhance material, nonlinear response. Traditional mirrors include metallic films and distributed Bragg reflectors. In this paper we propose negative index material mirrors as a third alternative. With the help of a rigorous Green function approach, we investigate second harmonic generation from single and coupled cavities, and theoretically prove that negative index material mirrors can raise the nonlinear conversion efficiency of a bulk material by at least four orders of magnitude compared to a bulk medium.

Non-critical phase-matching fourth harmonic generation of a 1053-nm laser in an ADP crystal

PubMed Central

Ji, Shaohua; Wang, Fang; Zhu, Lili; Xu, Xinguang; Wang, Zhengping; Sun, Xun

2013-01-01

In current inertial confinement fusion (ICF) facilities, KDP and DKDP crystals are the second harmonic generation (SHG) and third harmonic generation (THG) materials for the Nd:glass laser (1053 nm). Based on the trend for the development of short wavelengths for ICF driving lasers, technical solutions for fourth harmonic generation (FHG) will undoubtedly attract more and more attention. In this paper, the rapid growth of an ADP crystal and non-critical phase-matching (NCPM) FHG of a 1053-nm laser using an ADP crystal are reported. The NCPM temperature is 33.7°C. The conversion efficiency from 526 to 263 nm is 70%, and the angular acceptance range is 55.4 mrad; these results are superior to those for the DKDP crystals. This research has shown that ADP crystals will be a competitive candidate in future ICF facilities when the utilisation of high-energy, high-efficiency UV lasers at wavelengths shorter than the present 351 nm is of interest. PMID:23549389

Non-critical phase-matching fourth harmonic generation of a 1053-nm laser in an ADP crystal.

PubMed

Ji, Shaohua; Wang, Fang; Zhu, Lili; Xu, Xinguang; Wang, Zhengping; Sun, Xun

2013-01-01

In current inertial confinement fusion (ICF) facilities, KDP and DKDP crystals are the second harmonic generation (SHG) and third harmonic generation (THG) materials for the Nd:glass laser (1053 nm). Based on the trend for the development of short wavelengths for ICF driving lasers, technical solutions for fourth harmonic generation (FHG) will undoubtedly attract more and more attention. In this paper, the rapid growth of an ADP crystal and non-critical phase-matching (NCPM) FHG of a 1053-nm laser using an ADP crystal are reported. The NCPM temperature is 33.7°C. The conversion efficiency from 526 to 263 nm is 70%, and the angular acceptance range is 55.4 mrad; these results are superior to those for the DKDP crystals. This research has shown that ADP crystals will be a competitive candidate in future ICF facilities when the utilisation of high-energy, high-efficiency UV lasers at wavelengths shorter than the present 351 nm is of interest.

In vivo multiphoton microscopy of deep tissue with gradient index lenses

NASA Astrophysics Data System (ADS)

Levene, Michael J.; Dombeck, Daniel A.; Williams, Rebecca M.; Skoch, Jesse; Hickey, Gregory A.; Kasischke, Karl A.; Molloy, Raymond P.; Ingelsson, Martin; Stern, Edward A.; Klucken, Jochen; Bacskai, Brian J.; Zipfel, Warren R.; Hyman, Bradley T.; Webb, Watt W.

2004-06-01

Gradient index lenses enable multiphoton microscopy of deep tissues in the intact animal. In order to assess their applicability to clinical research, we present in vivo multiphoton microscopy with gradient index lenses in brain regions associated with Alzheimer's disease and Parkinson's disease in both transgenic and wild-type mice. We also demonstrate microscopy of ovary in wild type mouse using only intrinsic fluorescence and second harmonic generation, signal sources which may prove useful for both the study and diagnosis of cancer.

Higher-order Kerr effect and harmonic cascading in gases.

PubMed

Bache, Morten; Eilenberger, Falk; Minardi, Stefano

2012-11-15

The higher-order Kerr effect (HOKE) has recently been advocated to explain measurements of the saturation of the nonlinear refractive index in gases. Here we show that cascaded third-harmonic generation results in an effective fifth-order nonlinearity that is negative and significant. Higher-order harmonic cascading will also occur from the HOKE, and the cascading contributions may significantly modify the observed nonlinear index change. At lower wavelengths, cascading increases the HOKE saturation intensity, while for longer wavelengths cascading will decrease the HOKE saturation intensity.

Towards protein-crystal centering using second-harmonic generation (SHG) microscopy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kissick, David J.; Dettmar, Christopher M.; Becker, Michael

2013-05-01

The potential of second-harmonic generation (SHG) microscopy for automated crystal centering to guide synchrotron X-ray diffraction of protein crystals has been explored. The potential of second-harmonic generation (SHG) microscopy for automated crystal centering to guide synchrotron X-ray diffraction of protein crystals was explored. These studies included (i) comparison of microcrystal positions in cryoloops as determined by SHG imaging and by X-ray diffraction rastering and (ii) X-ray structure determinations of selected proteins to investigate the potential for laser-induced damage from SHG imaging. In studies using β{sub 2} adrenergic receptor membrane-protein crystals prepared in lipidic mesophase, the crystal locations identified by SHGmore » images obtained in transmission mode were found to correlate well with the crystal locations identified by raster scanning using an X-ray minibeam. SHG imaging was found to provide about 2 µm spatial resolution and shorter image-acquisition times. The general insensitivity of SHG images to optical scatter enabled the reliable identification of microcrystals within opaque cryocooled lipidic mesophases that were not identified by conventional bright-field imaging. The potential impact of extended exposure of protein crystals to five times a typical imaging dose from an ultrafast laser source was also assessed. Measurements of myoglobin and thaumatin crystals resulted in no statistically significant differences between structures obtained from diffraction data acquired from exposed and unexposed regions of single crystals. Practical constraints for integrating SHG imaging into an active beamline for routine automated crystal centering are discussed.« less

Generation of coherent terahertz radiation in ultrafast laser-gas interactionsa)

NASA Astrophysics Data System (ADS)

Kim, Ki-Yong

2009-05-01

The generation of intense terahertz radiation in ultrafast laser-gas interactions is studied on a basis of transient electron current model. When an ultrashort pulse laser's fundamental and its second harmonic fields are mixed to ionize a gas, a nonvanishing, directional photoelectron current can be produced, which simultaneously emits terahertz radiation in the far field. Here, the generation mechanism is examined with an analytic derivation and numerical simulations, in which tunneling ionization and subsequent electron motion in the combined laser field play a key role. In the simulations, three types of laser-gas interactions are considered: (i) mixing the fundamental and its second harmonic fields, (ii) mixing nonharmonic, two-color fields, and (iii) focusing single-color, few-cycle pulses. In these interactions, terahertz generation and other nonlinear effects driven by the transient current are investigated. In particular, anticorrelation between terahertz and second (or third) harmonic generation is observed and analyzed.

Anisotropic high-harmonic generation in bulk crystals

DOE Office of Scientific and Technical Information (OSTI.GOV)

You, Yong Sing; Reis, David A.; Ghimire, Shambhu

2016-11-21

The microscopic valence electron density determines the optical, electronic, structural and thermal properties of materials. However, current techniques for measuring this electron charge density are limited: for example, scanning tunnelling microscopy is confined to investigations at the surface, and electron diffraction requires very thin samples to avoid multiple scattering. Therefore, an optical method is desirable for measuring the valence charge density of bulk materials. Since the discovery of high-harmonic generation (HHG) in solids, there has been growing interest in using HHG to probe the electronic structure of solids. Here, using single-crystal MgO, we demonstrate that high-harmonic generation in solids ismore » sensitive to interatomic bonding. We find that harmonic efficiency is enhanced (diminished) for semi-classical electron trajectories that connect (avoid) neighbouring atomic sites in the crystal. Finally, these results indicate the possibility of using materials’ own electrons for retrieving the interatomic potential and thus the valence electron density, and perhaps even wavefunctions, in an all-optical setting.« less

Rigorous theory of molecular orientational nonlinear optics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kwak, Chong Hoon, E-mail: chkwak@ynu.ac.kr; Kim, Gun Yeup

2015-01-15

Classical statistical mechanics of the molecular optics theory proposed by Buckingham [A. D. Buckingham and J. A. Pople, Proc. Phys. Soc. A 68, 905 (1955)] has been extended to describe the field induced molecular orientational polarization effects on nonlinear optics. In this paper, we present the generalized molecular orientational nonlinear optical processes (MONLO) through the calculation of the classical orientational averaging using the Boltzmann type time-averaged orientational interaction energy in the randomly oriented molecular system under the influence of applied electric fields. The focal points of the calculation are (1) the derivation of rigorous tensorial components of the effective molecularmore » hyperpolarizabilities, (2) the molecular orientational polarizations and the electronic polarizations including the well-known third-order dc polarization, dc electric field induced Kerr effect (dc Kerr effect), optical Kerr effect (OKE), dc electric field induced second harmonic generation (EFISH), degenerate four wave mixing (DFWM) and third harmonic generation (THG). We also present some of the new predictive MONLO processes. For second-order MONLO, second-order optical rectification (SOR), Pockels effect and difference frequency generation (DFG) are described in terms of the anisotropic coefficients of first hyperpolarizability. And, for third-order MONLO, third-order optical rectification (TOR), dc electric field induced difference frequency generation (EFIDFG) and pump-probe transmission are presented.« less

Monitoring microstructural evolution of alloy 617 with non-linear acoustics for remaining useful life prediction; multiaxial creep-fatigue and creep-ratcheting

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lissenden, Cliff; Hassan, Tasnin; Rangari, Vijaya

The research built upon a prior investigation to develop a unified constitutive model for design-­by-­analysis of the intermediate heat exchanger (IHX) for a very high temperature reactor (VHTR) design of next generation nuclear plants (NGNPs). Model development requires a set of failure data from complex mechanical experiments to characterize the material behavior. Therefore uniaxial and multiaxial creep-­fatigue and creep-­ratcheting tests were conducted on the nickel-­base Alloy 617 at 850 and 950°C. The time dependence of material behavior, and the interaction of time dependent behavior (e.g., creep) with ratcheting, which is an increase in the cyclic mean strain under load-­controlled cycling,more » are major concerns for NGNP design. This research project aimed at characterizing the microstructure evolution mechanisms activated in Alloy 617 by mechanical loading and dwell times at elevated temperature. The acoustic harmonic generation method was researched for microstructural characterization. It is a nonlinear acoustics method with excellent potential for nondestructive evaluation, and even online continuous monitoring once high temperature sensors become available. It is unique because it has the ability to quantitatively characterize microstructural features well before macroscale defects (e.g., cracks) form. The nonlinear acoustics beta parameter was shown to correlate with microstructural evolution using a systematic approach to handle the complexity of multiaxial creep-­fatigue and creep-­ratcheting deformation. Mechanical testing was conducted to provide a full spectrum of data for: thermal aging, tensile creep, uniaxial fatigue, uniaxial creep-­fatigue, uniaxial creep-ratcheting, multiaxial creep-fatigue, and multiaxial creep-­ratcheting. Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM), and Optical Microscopy were conducted to correlate the beta parameter with individual microstructure mechanisms. We researched application of the harmonic generation method to tubular mechanical test specimens and pipes for nondestructive evaluation. Tubular specimens and pipes act as waveguides, thus we applied the acoustic harmonic generation method to guided waves in both plates and shells. Magnetostrictive transducers were used to generate and receive guided wave modes in the shell sample and the received signals were processed to show the sensitivity of higher harmonic generation to microstructure evolution. Modeling was initiated to correlate higher harmonic generation with the microstructure that will lead to development of a life prediction model that is informed by the nonlinear acoustics measurements.« less

Soliton self-frequency shift and third-harmonic generation in a four-hole As₂S₅ microstructured optical fiber.

PubMed

Cheng, Tonglei; Usaki, Ryo; Duan, Zhongchao; Gao, Weiqing; Deng, Dinghuan; Liao, Meisong; Kanou, Yasuhire; Matsumoto, Morio; Misumi, Takashi; Suzuki, Takenobu; Ohishi, Yasutake

2014-02-24

Soliton self-frequency shift (SSFS) and third-harmonic generation (THG) are observed in a four-hole As2S5 chalcogenide microstructured optical fiber (MOF). The As2S5 MOF is tapered to offer an ideal environment for SSFS. After tapering, the zero-dispersion wavelength (ZDW) shifts from 2.02 to 1.61 μm, and the rate of SSFS can be enhanced by increasing the energy density of the pulse. By varying the average input power from 220 to 340 mW, SSFS of a soliton central wavelength from 2.206 to 2.600 μm in the mid-infrared is observed in the tapered segment, and THG at 632 nm is observed in the untapered segment.

Tumor tissue characterization using polarization-sensitive second harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Tokarz, Danielle; Cisek, Richard; Golaraei, Ahmad; Krouglov, Serguei; Navab, Roya; Niu, Carolyn; Sakashita, Shingo; Yasufuku, Kazuhiro; Tsao, Ming-Sound; Asa, Sylvia L.; Barzda, Virginijus; Wilson, Brian C.

2015-06-01

Changes in the ultrastructure of collagen in various tumor and non-tumor human tissues including lung, pancreas and thyroid were investigated ex vivo by a polarization-sensitive second harmonic generation (SHG) microscopy technique referred to as polarization-in, polarization-out (PIPO) SHG. This involves measuring the orientation of the linear polarization of outgoing SHG as a function of the linear polarization orientation of incident laser radiation. From the PIPO SHG data, the second-order nonlinear optical susceptibility tensor component ratio, χ(2) ZZZ'/χ(2) ZXX', for each pixel of the SHG image was obtained and presented as color-coded maps. Further, the orientation of collagen fibers in the tissue was deduced. Since the χ(2) ZZZ'/χ(2) ZXX' values represent the organization of collagen in the tissue, theses maps revealed areas of altered collagen structure (not simply concentration) within tissue sections. Statistically-significant differences in χ(2) ZZZ'/χ(2) ZXX' were found between tumor and non-tumor tissues, which varied from organ to organ. Hence, PIPO SHG microscopy could potentially be used to aid pathologists in diagnosing cancer. Additionally, PIPO SHG microscopy could aid in characterizing the structure of collagen in other collagen-related biological processes such as wound repair.

Precise, motion-free polarization control in Second Harmonic Generation microscopy using a liquid crystal modulator in the infinity space.

PubMed

Lien, Chi-Hsiang; Tilbury, Karissa; Chen, Shean-Jen; Campagnola, Paul J

2013-01-01

Second Harmonic Generation (SHG) microscopy coupled with polarization analysis has great potential for use in tissue characterization, as molecular and supramolecular structural details can be extracted. Such measurements are difficult to perform quickly and accurately. Here we present a new method that uses a liquid crystal modulator (LCM) located in the infinity space of a SHG laser scanning microscope that allows the generation of any desired linear or circular polarization state. As the device contains no moving parts, polarization can be rotated accurately and faster than by manual or motorized control. The performance in terms of polarization purity was validated using Stokes vector polarimetry, and found to have minimal residual polarization ellipticity. SHG polarization imaging characteristics were validated against well-characterized specimens having cylindrical and/or linear symmetries. The LCM has a small footprint and can be implemented easily in any standard microscope and is cost effective relative to other technologies.

Precise, motion-free polarization control in Second Harmonic Generation microscopy using a liquid crystal modulator in the infinity space

PubMed Central

Lien, Chi-Hsiang; Tilbury, Karissa; Chen, Shean-Jen; Campagnola, Paul J.

2013-01-01

Second Harmonic Generation (SHG) microscopy coupled with polarization analysis has great potential for use in tissue characterization, as molecular and supramolecular structural details can be extracted. Such measurements are difficult to perform quickly and accurately. Here we present a new method that uses a liquid crystal modulator (LCM) located in the infinity space of a SHG laser scanning microscope that allows the generation of any desired linear or circular polarization state. As the device contains no moving parts, polarization can be rotated accurately and faster than by manual or motorized control. The performance in terms of polarization purity was validated using Stokes vector polarimetry, and found to have minimal residual polarization ellipticity. SHG polarization imaging characteristics were validated against well-characterized specimens having cylindrical and/or linear symmetries. The LCM has a small footprint and can be implemented easily in any standard microscope and is cost effective relative to other technologies. PMID:24156059

Effect of the magnetic field on the nonlinear optical rectification and second and third harmonic generation in double δ-doped GaAs quantum wells

NASA Astrophysics Data System (ADS)

Martínez-Orozco, J. C.; Rojas-Briseño, J. G.; Rodríguez-Magdaleno, K. A.; Rodríguez-Vargas, I.; Mora-Ramos, M. E.; Restrepo, R. L.; Ungan, F.; Kasapoglu, E.; Duque, C. A.

2017-11-01

In this paper we are reporting the computation for the Nonlinear Optical Rectification (NOR) and the Second and Third Harmonic Generation (SHG and THG) related with electronic states of asymmetric double Si-δ-doped quantum well in a GaAs matrix when this is subjected to an in-plane (x-oriented) constant magnetic field effect. The work is performed in the effective mass and parabolic band approximations in order to compute the electronic structure for the system by a diagonalization procedure. The expressions for the nonlinear optical susceptibilities, χ0(2), χ2ω(2), and χ3ω(3), are those arising from the compact matrix density formulation and stand for the NOR, SHG, and THG, respectively. This asymmetric double δ-doped quantum well potential profile actually exhibits nonzero NOR, SHG, and THG responses which can be easily controlled by the in-plane (x-direction) externally applied magnetic field. In particular we find that for the chosen configuration the harmonic generation is in the far-infrared/THz region, thus and becoming suitable building blocks for photodetectors in this range of the electromagnetic spectra.

Resonantly enhanced method for generation of tunable, coherent vacuum ultraviolet radiation

DOEpatents

Glownia, James H.; Sander, Robert K.

1985-01-01

Carbon Monoxide vapor is used to generate coherent, tunable vacuum ultraviolet radiation by third-harmonic generation using a single tunable dye laser. The presence of a nearby electronic level resonantly enhances the nonlinear susceptibility of this molecule allowing efficient generation of the vuv light at modest pump laser intensities, thereby reducing the importance of a six-photon multiple-photon ionization process which is also resonantly enhanced by the same electronic level but to higher order. By choosing the pump radiation wavelength to be of shorter wavelength than individual vibronic levels used to extend tunability stepwise from 154.4 to 124.6 nm, and the intensity to be low enough, multiple-photon ionization can be eliminated. Excitation spectra of the third-harmonic emission output exhibit shifts to shorter wavelength and broadening with increasing CO pressure due to phase matching effects. Increasing the carbon monoxide pressure, therefore, allows the substantial filling in of gaps arising from the stepwise tuning thereby providing almost continuous tunability over the quoted range of wavelength emitted.

Resonantly enhanced method for generation of tunable, coherent vacuum-ultraviolet radiation

DOEpatents

Glownia, J.H.; Sander, R.K.

1982-06-29

Carbon Monoxide vapor is used to generate coherent, tunable vacuum ultraviolet radiation by third-harmonic generation using a single tunable dye laser. The presence of a nearby electronic level resonantly enhances the nonlinear susceptibility of this molecule allowing efficient generation of the vuv light at modest pump laser intensities, thereby reducing the importance of a six-photon multiple-photon ionization process which is also resonantly enhanced by the same electronic level but no higher order. By choosing the pump radiation wavelength to be of shorter wavelength than individual vibronic levels used to extend tunability stepwise from 154.4 to 124.6 nm, and the intensity to be low enough, multiple-photon ionization can be eliminated. Excitation spectra of the third-harmonic emission output exhibit shifts to shorter wavelength and broadening with increasing CO pressure due to phase matching effects. Increasing the carbon monoxide pressure, therefore, allows the substantial filling in of gaps arising from the stepwise tuning thereby providing almost continuous tunability over the quoted range of wavelength emitted.

Theory of plasmonic effects in nonlinear optics: the case of graphene

NASA Astrophysics Data System (ADS)

Rostami, Habib; Katsnelson, Mikhail I.; Polini, Marco; Mikhail I. Katsnelson Collaboration; Habib Rostami; Marco Polini Collaboration

The nonlinear optical properties of two-dimensional electronic systems are beginning to attract considerable interest both in the theoretical and experimental sectors. Recent experiments on the nonlinear optical properties of graphene reveal considerably strong third harmonic generation and four-wave mixing of this single-atomic-layer electronic system. We develop a large-N theory of electron-electron interaction corrections to multi-legged Feynman diagrams describing second- and third-order nonlinear response functions. Our theory is completely general and is useful to understand all second- and third-order nonlinear effects, including harmonic generation, wave mixing, and photon drag. We apply our theoretical framework to the case of graphene, by carrying out microscopic calculations of the second- and third-order nonlinear response functions of an interacting two-dimensional gas of massless Dirac fermions. We compare our results with recent measurements, where all-optical launching of graphene plasmons has been achieved. This work was supported by Fondazione Istituto Italiano di Tecnologia, the European Union's Horizon 2020 research and innovation programme under Grant agreement No. 696656 GrapheneCore, and the ERC Advanced Grant 338957 FEMTO/NANO (M.I.K.).

Hyperspectral stimulated Raman scattering and multiphoton imaging for digital pathology of colonic disease

NASA Astrophysics Data System (ADS)

Wang, Zi; Zheng, Wei; Lin, Jian; Huang, Zhiwei

2016-03-01

Histopathology examinations of H&E stained biopsied tissues is the golden standard for colonic diseases (e.g., polyps, adenoma, and adenocarcinoma) diagnosis. However, staining effect of sample and doctor's expertise degree may greatly influence the diagnosis results. The information provided by the H&E stained sample is also limited to the morphological and PH information and no quantative information is available. In this paper, we report the development of a unique multimodal nonlinear optical microscopy (i.e., hyperspectral stimulated Raman scattering (hsSRS), second-harmonic generation (SHG), third-harmonic generation (THG), two-photon excitation fluorescence (TPEF)) platform for the diagnosis and characterization of colonic diseases. HsSRS in both fingerprint (800-1800 cm-1) and high-wavenumber (2800-3600 cm-1) regions allows us to discriminate different constituents with tiny difference in the Raman spectra. The increase of proteins and reduction of lipids could be observed with the progress of colonic cancer. SHG shows the distribution of collagen, which is found to aggregate for adenocarcinoma. TPEF provides the cell morphological and can reflect the damage inside glands caused by the diseases. THG shows the increase of optical heterogeneity related to cancer process. This work shows that the integrated hsSRS and TPEF/SHG/THG imaging technique can be an effective method for digital pathology of colonic diseases at the molecular and sub-cellular levels.

Second-harmonic generation microscopy of tooth

NASA Astrophysics Data System (ADS)

Kao, Fu-Jen; Wang, Yung-Shun; Huang, Mao-Kuo; Huang, Sheng-Lung; Cheng, Ping C.

2000-07-01

In this study, we have developed a high performance microscopic system to perform second-harmonic (SH)imaging on a tooth. The high sensitivity of the system allows an acquisition rate of 300 seconds/frame with a resolution at 512x512 pixels. The surface SH signal generated from the tooth is also carefully verified through micro-spectroscopy, polarization rotation, and wavelength tuning. In this way, we can ensure the authenticity of the signal. The enamel that encapsulates the dentine is known to possess highly ordered structures. The anisotrophy of the structure is revealed in the microscopic SH images of the tooth sample.

Polarization-resolved second-harmonic generation microscopy as a method to visualize protein-crystal domains

PubMed Central

DeWalt, Emma L.; Begue, Victoria J.; Ronau, Judith A.; Sullivan, Shane Z.; Das, Chittaranjan; Simpson, Garth J.

2013-01-01

Polarization-resolved second-harmonic generation (PR-SHG) microscopy is described and applied to identify the presence of multiple crystallographic domains within protein-crystal conglomerates, which was confirmed by synchrotron X-ray diffraction. Principal component analysis (PCA) of PR-SHG images resulted in principal component 2 (PC2) images with areas of contrasting negative and positive values for conglomerated crystals and PC2 images exhibiting uniformly positive or uniformly negative values for single crystals. Qualitative assessment of PC2 images allowed the identification of domains of different internal ordering within protein-crystal samples as well as differentiation between multi-domain conglomerated crystals and single crystals. PR-SHG assessments of crystalline domains were in good agreement with spatially resolved synchrotron X-ray diffraction measurements. These results have implications for improving the productive throughput of protein structure determination through early identification of multi-domain crystals. PMID:23275165

Polarization-resolved second-harmonic generation imaging for liver fibrosis assessment without labeling

NASA Astrophysics Data System (ADS)

Lin, Jian; Pan, Shiying; Zheng, Wei; Huang, Zhiwei

2013-10-01

We apply the polarization-resolved second-harmonic generation (PR-SHG) microscopy to investigate the changes of collagen typings (type I vs type III) and collagen fibril orientations of liver tissue in bile-duct-ligation (BDL) rat models. The PR-SHG results show that the second-order susceptibility tensor ratios (χ31/χ15 and χ33/χ15) of collagen fibers increase with liver fibrotic progression after BDL surgery, reflecting an increase of the type III collagen component with the severity of liver fibrosis; and the square root of the collagen type III to type I ratio linearly correlates (R2 = 0.98) with histopathological scores. Furthermore, the collagen fibril orientations become more random with liver fibrosis transformation as compared to normal liver tissue. This work demonstrates that PR-SHG microscopy has the potential for label-free diagnosis and characterization of liver fibrosis based on quantitative analysis of collagen typings and fibril orientations.

Quantifying collagen orientation in breast tissue biopsies using SLIM (Conference Presentation)

NASA Astrophysics Data System (ADS)

Majeed, Hassaan; Okoro, Chukwuemeka; Balla, Andre; Toussaint, Kimani C.; Popescu, Gabriel

2017-02-01

Breast cancer is a major public health problem worldwide, being the most common type of cancer among women according to the World Health Organization (WHO). The WHO has further stressed the importance of an early determination of the disease course through prognostic markers. Recent studies have shown that the alignment of collagen fibers in tumor adjacent stroma correlate with poorer health outcomes in patients. Such studies have typically been carried out using Second-Harmonic Generation (SHG) microscopy. SHG images are very useful for quantifying collagen fiber orientation due their specificity to non-centrosymmetric structures in tissue, leading to high contrast in collagen rich areas. However, the imaging throughput in SHG microscopy is limited by its point scanning geometry. In this work, we show that SLIM, a wide-field high-throughput QPI technique, can be used to obtain the same information on collagen fiber orientation as is obtainable through SHG microscopy. We imaged a tissue microarray containing both benign and malignant cores using both SHG microscopy and SLIM. The cellular (non-collagenous) structures in the SLIM images were next segmented out using an algorithm developed in-house. Using the previously published Fourier Transform Second Harmonic Generation (FT-SHG) tool, the fiber orientations in SHG and segmented SLIM images were then quantified. The resulting histograms of fiber orientation angles showed that both SHG and SLIM generate similar measurements of collagen fiber orientation. The SLIM modality, however, can generate these results at much higher throughput due to its wide-field, whole-slide scanning capabilities.

Analysis of human knee osteoarthritic cartilage using polarization sensitive second harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Kumar, Rajesh; Grønhaug, Kirsten M.; Romijn, Elisabeth I.; Drogset, Jon O.; Lilledahl, Magnus B.

2014-05-01

Osteoarthritis is one of the most prevalent joint diseases in the world. Although the cause of osteoarthritis is not exactly clear, the disease results in a degradation of the quality of the articular cartilage including collagen and other extracellular matrix components. We have investigated alterations in the structure of collagen fibers in the cartilage tissue of the human knee using mulitphoton microscopy. Due to inherent high nonlinear susceptibility, ordered collagen fibers present in the cartilage tissue matrix produces strong second harmonic generation (SHG) signals. Significant morphological differences are found in different Osteoarthritic grades of cartilage by SHG microscopy. Based on the polarization analysis of the SHG signal, we find that a few locations of hyaline cartilage (mainly type II collagen) is being replaced by fibrocartilage (mainly type I cartilage), in agreement with earlier literature. To locate the different types and quantify the alteration in the structure of collagen fiber, we employ polarization-SHG microscopic analysis, also referred to as _-tensor imaging. The image analysis of p-SHG image obtained by excitation polarization measurements would represent different tissue constituents with different numerical values at pixel level resolution.

Nonlinear microscopy of collagen fibers

NASA Astrophysics Data System (ADS)

Strupler, M.; Pena, A.-M.; Hernest, M.; Tharaux, P.-L.; Fabre, A.; Marchal-Somme, J.; Crestani, B.; Débarre, D.; Martin, J.-L.; Beaurepaire, E.; Schanne-Klein, M.-C.

2007-02-01

We used intrinsic Second Harmonic Generation (SHG) by fibrillar collagen to visualize the three-dimensional architecture of collagen fibrosis at the micrometer scale using laser scanning nonlinear microscopy. We showed that SHG signals are highly specific to fibrillar collagen and provide a sensitive probe of the micrometer-scale structural organization of collagen in tissues. Moreover, recording simultaneously other nonlinear optical signals in a multimodal setup, we visualized the tissue morphology using Two-Photon Excited Fluorescence (2PEF) signals from endogenous chromophores such as NADH or elastin. We then compared different methods to determine accurate indexes of collagen fibrosis using nonlinear microscopy, given that most collagen fibrils are smaller than the microscope resolution and that second harmonic generation is a coherent process. In order to define a robust method to process our three-dimensional images, we either calculated the fraction of the images occupied by a significant SHG signal, or averaged SHG signal intensities. We showed that these scores provide an estimation of the extension of renal and pulmonary fibrosis in murine models, and that they clearly sort out the fibrotic mice.

Robust generation of Fourier-synthesized laser fields and their estimation of the optical phase by using quantum control of molecular tunneling ionization

NASA Astrophysics Data System (ADS)

Yoshida, Tsuyoshi; Saito, Naoaki; Ohmura, Hideki

2018-03-01

Intense (5.0 × 1012 W cm-2) nanosecond Fourier-synthesized laser fields consisting of fundamental, second-, third-, and fourth-harmonic light generated by an interferometer-free Fourier-synthesized laser field generator induce orientation-selective ionization based on directionally asymmetric molecular tunneling ionization (TI). The laser field generator ensures adjustment-free operation, high stability, and high reproducibility. Phase-sensitive, orientation-selective molecular TI provides a simple way to estimate the relative phase differences between the fundamental light and each harmonic by data-fitting analysis. This application of Fourier-synthesized laser fields will facilitate not only lightwave engineering but also the control of matter.

Investigation of Optical Cavity Modes and Ultrafast Carrier Dynamics in Zinc Oxide Rods Using Second-Harmonic Generation and Transient Absorption Pump-Probe Microscopy

NASA Astrophysics Data System (ADS)

Mehl, Brian Peter

The polydispersity intrinsic to nanoscale and microscale semiconductor materials poses a major challenge to using individual objects as building blocks for device applications. The ability to manipulate the shape of ZnO structures is enormous, making it an ideal material for studying shape-dependent phenomena. We have built a nonlinear microscope used to directly image optical cavity modes in ZnO rods using second-harmonic generation. Images of second-harmonic generation in needle-shaped ZnO rods obtained from individual structures show areas of enhanced second-harmonic intensity along the longitudinal axis of the rod that are periodically distributed and symmetrically situated relative to the rod midpoint. The spatial modulation is a direct consequence of the fundamental optical field coupling into standing wave resonator modes of the ZnO structure, leading to an enhanced backscattered second-harmonic condition that cannot be achieved in bulk ZnO. A more complicated second-harmonic image is observed when excitation is below the band gap, which is attributed to whispering gallery modes. Additionally, the nonlinear microscope was combined with transient absorption pump-probe to follow the electron-hole recombination dynamics at different points within individual needle-shaped ZnO rods to characterize spatial differences in dynamical behavior. The results from pump-probe experiments are correlated with spatially resolved ultrafast emission measurements, and scanning electron microscopy provides structural details. Dramatically different electron-hole recombination dynamics are observed in the narrow tips compared to the interior, with the ends exhibiting a greater propensity for electron-hole plasma formation and faster recombination of carriers across the band gap that stem from a physical confinement of the charge carriers. In the interior of the rod, a greater fraction of the electron-hole recombination is trap-mediated and occurs on a significantly longer time scale.

CARS molecular fingerprinting using a sub-nanosecond supercontinuum light source

NASA Astrophysics Data System (ADS)

Kano, Hideaki; Akiyama, Toshihiro; Inoko, Akihito; Kobayashi, Tsubasa; Leproux, Philippe; Couderc, Vincent; Kaji, Yuichi; Oshika, Tetsuro

2018-02-01

We have visualized living cells and tissues using an ultrabroadband multiplex coherent anti-Stokes Raman scattering (CARS) microspectroscopic system by using a sub-nanosecond supercontinuum (SC) light source. Owing to the ultrabroadband spectral profile of the SC, we can generate multiplex CARS signals in the spectral range of 500-3800 cm-1, which covers the whole molecular fingerprint region, as well as the C-H and O-H stretching regions. Through the combination of the ultrabroadband multiplex CARS method with second harmonic generation (SHG) and third harmonic generation (THG) processes, we have successfully performed selective imaging of ciliary rootlet-composing Rootletin filaments in rat retina.

Resonant third harmonic generation of KrF laser in Ar gas.

PubMed

Rakowski, R; Barna, A; Suta, T; Bohus, J; Földes, I B; Szatmári, S; Mikołajczyk, J; Bartnik, A; Fiedorowicz, H; Verona, C; Verona Rinati, G; Margarone, D; Nowak, T; Rosiński, M; Ryć, L

2014-12-01

Investigations of emission of harmonics from argon gas jet irradiated by 700 fs, 5 mJ pulses from a KrF laser are presented. Harmonics conversion was optimized by varying the experimental geometry and the nozzle size. For the collection of the harmonic radiation silicon and solar-blind diamond semiconductor detectors equipped with charge preamplifiers were applied. The possibility of using a single-crystal CVD diamond detector for separate measurement of the 3rd harmonic in the presence of a strong pumping radiation was explored. Our experiments show that the earlier suggested 0.7% conversion efficiency can really be obtained, but only in the case when phase matching is optimized with an elongated gas target length corresponding to the length of coherence.

Using surface lattice resonances to engineer nonlinear optical processes in metal nanoparticle arrays

NASA Astrophysics Data System (ADS)

Huttunen, Mikko J.; Rasekh, Payman; Boyd, Robert W.; Dolgaleva, Ksenia

2018-05-01

Collective responses of localized surface plasmon resonances, known as surface lattice resonances (SLRs) in metal nanoparticle arrays, can lead to high quality factors (˜100 ), large local-field enhancements, and strong light-matter interactions. SLRs have found many applications in linear optics, but little work of the influence of SLRs on nonlinear optics has been reported. Here we show how SLRs could be utilized to enhance nonlinear optical interactions. We devote special attention to the sum-frequency, difference-frequency, and third-harmonic generation processes because of their potential for the realization of novel sources of light. We also demonstrate how such arrays could be engineered to enhance higher-order nonlinear optical interactions through cascaded nonlinear processes. In particular, we demonstrate how the efficiency of third-harmonic generation could be engineered via cascaded second-order responses.

Impact of the phase-mismatch in the SHG crystal and consequential self-action of the fundamental wave by cascaded second-order effects on the THG efficiency of a Q-switched 1342 nm Nd:YVO₄ laser.

PubMed

Koch, Peter; Bartschke, Juergen; L'huillier, Johannes A

2015-05-18

We report on the influence of self-focusing and self-defocusing in the phase-mismatched frequency doubling crystal on the third harmonic generation (THG) efficiency in a two crystal frequency tripling scheme. By detuning the temperature of the doubling crystal, the impact of a phase-mismatch in second harmonic generation (SHG) on the subsequent sum frequency mixing process was investigated. It was found that adjusting the temperature not only affected the power ratio of the second harmonic to the fundamental but also the beam diameter of the fundamental beam in the THG crystal, which was caused by self-focusing and self-defocusing of the fundamental beam, respectively. This self-action was induced by a cascaded χ(2) : χ(2) process in the phase-mismatched SHG crystal. Self-defocusing was observable for positive detuning and self-focusing for negative detuning of the phase-matching temperature. Hence, the THG efficiency was not symmetric with respect to the point of optimum phase-matching. Optimum THG was obtained for positive detuning and the resulting self-defocusing in combination with the focusing lens in front of the THG stage was also beneficial for the beam quality of the third harmonic.

On-target diagnosing of few-cycle pulses by high-order-harmonic generation

NASA Astrophysics Data System (ADS)

Brambila, Danilo S.; Husakou, Anton; Ivanov, Misha; Zhavoronkov, Nickolai

2017-12-01

We propose an approach to determine the residual phase distortion directly in the interaction region of few-cycle laser radiation with a gaseous target. We describe how the spectra of the generated high harmonics measured as a function of externally introduced dispersion into the driving few-cycle laser pulse can be used to decode small amounts of second- and third-order spectral phase, including the sign. The diagnosis is based on the analysis of several key features in the high-harmonic spectrum: the depth of spectral modulation, the position of the cutoff, and the symmetry of the spectrum with respect to the introduced dispersion. The approach is applicable to pulses without carrier-envelope phase (CEP) stabilization. Surprisingly, we find that for nearly-single-cycle pulses with nonstabilized CEP, deep spectral modulations in the harmonic spectra emerge for positively rather than negatively chirped pulses, in contrast to the case of CEP-stabilized pulses.

Optical nonlinearities of excitons in monolayer MoS2

NASA Astrophysics Data System (ADS)

Soh, Daniel B. S.; Rogers, Christopher; Gray, Dodd J.; Chatterjee, Eric; Mabuchi, Hideo

2018-04-01

We calculate linear and nonlinear optical susceptibilities arising from the excitonic states of monolayer MoS2 for in-plane light polarizations, using second-quantized bound and unbound exciton operators. Optical selection rules are critical for obtaining the susceptibilities. We derive the valley-chirality rule for the second-order harmonic generation in monolayer MoS2 and find that the third-order harmonic process is efficient only for linearly polarized input light while the third-order two-photon process (optical Kerr effect) is efficient for circularly polarized light using a higher order exciton state. The absence of linear absorption due to the band gap and the unusually strong two-photon third-order nonlinearity make the monolayer MoS2 excitonic structure a promising resource for coherent nonlinear photonics.

Quasi-phase-matched χ(3 )-parametric interactions in sinusoidally tapered waveguides

NASA Astrophysics Data System (ADS)

Saleh, Mohammed F.

2018-01-01

In this article, I show how periodically tapered waveguides can be employed as efficient quasi-phase-matching schemes for four-wave mixing parametric processes in third-order nonlinear materials. As an example, a thorough study of enhancing third-harmonic generation in sinusoidally tapered fibers has been conducted. The quasi-phase-matching condition has been obtained for nonlinear parametric interactions in these structures using Fourier-series analysis. The dependencies of the conversion efficiency of the third harmonic on the modulation amplitude, tapering period, longitudinal-propagation direction, and pump wavelength have been studied. In comparison to uniform waveguides, the conversion efficiency has been enhanced by orders of magnitudes. I envisage that this work will have a great impact in the field of guided nonlinear optics using centrosymmetric materials.

Nonlinear plasmonic imaging techniques and their biological applications

NASA Astrophysics Data System (ADS)

Deka, Gitanjal; Sun, Chi-Kuang; Fujita, Katsumasa; Chu, Shi-Wei

2017-01-01

Nonlinear optics, when combined with microscopy, is known to provide advantages including novel contrast, deep tissue observation, and minimal invasiveness. In addition, special nonlinearities, such as switch on/off and saturation, can enhance the spatial resolution below the diffraction limit, revolutionizing the field of optical microscopy. These nonlinear imaging techniques are extremely useful for biological studies on various scales from molecules to cells to tissues. Nevertheless, in most cases, nonlinear optical interaction requires strong illumination, typically at least gigawatts per square centimeter intensity. Such strong illumination can cause significant phototoxicity or even photodamage to fragile biological samples. Therefore, it is highly desirable to find mechanisms that allow the reduction of illumination intensity. Surface plasmon, which is the collective oscillation of electrons in metal under light excitation, is capable of significantly enhancing the local field around the metal nanostructures and thus boosting up the efficiency of nonlinear optical interactions of the surrounding materials or of the metal itself. In this mini-review, we discuss the recent progress of plasmonics in nonlinear optical microscopy with a special focus on biological applications. The advancement of nonlinear imaging modalities (including incoherent/coherent Raman scattering, two/three-photon luminescence, and second/third harmonic generations that have been amalgamated with plasmonics), as well as the novel subdiffraction limit imaging techniques based on nonlinear behaviors of plasmonic scattering, is addressed.

IR Bandwidth and Crystal Thickness Effects on THG Efficiency and Temporal Shaping of Quasi-Rectangular UV Pulses: Part II - Incident IR Ripple

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bolton, Paul R.; Limborg-Deprey, Cecile; /SLAC

We have investigated the effect of incident ir spectral bandwidth and crystal thickness, on uv pulses produced by third harmonic generation (THG) in a crystal pair. Our focus is on the third harmonic generation efficiency and longitudinal uv intensity profile parameters of SNLO predictions that are evaluated for three incident ir spectral bandwidths and a range of crystal thicknesses. These results represent a continuation of earlier work in which the effects of the same selected ir bandwidths and range of crystal thicknesses were investigated using a pair of BBO Type I crystals in a simplistic geometry for which the longitudinalmore » intensity plateau has a zero slope, 'flattop' profile. The current work is distinguished from the previous work by an imposed ripple on the ir intensity longitudinal profile and constitutes a portion of a Part II effort to which we have made reference. As with preceding work, all third harmonic data are net results at the exit of the second BBO crystal. Predictions are obtained with the modified SNLO code developed by Arlee Smith at the Sandia National Laboratories. This modification has allowed us to pursue the 'coupled' case in which the output of the first BBO crystal is used as input to the second one. This includes both the fundamental and second harmonic light. Defined parameters are consistent with previous work. The presented cases are best results. The criteria for selection of these reported cases are highest THG efficiency combined with minimum intensity ripple in the plateau. The incident ir pulse is quasi-rectangular with an imposed 5.2 % (rms) intensity ripple added to the plateau. The ir pulse bandwidth is centered at 800 nm. Second harmonic generation occurs in the first BBO crystal and THG occurs in the second crystal as a consequence of sum frequency generation. Type I phase matching is used throughout, so that for a negative uniaxial crystal: n{sub 2}{sup e}({theta}) = n{sub 1}{sup o};(SHG) (1.1) 3n{sub 3}{sup 3}({theta}) = 2n{sub 2}{sup o} + n{sub 1}{sup o};(THG) where n{sub 2}{sup e}({theta}) and n{sub 3}{sup e}({theta}) are the angle dependent extraordinary refractive indices for the second and third harmonics respectively, and n{sub 1}{sup 1} and n{sub 2}{sup o} are the ordinary refractive indices for the fundamental and second harmonic respectively. Although our goal at this stage has not been to comply with all the LCLS injector laser specifications, the results provided here represent a parameter study that can be used to determine candidate bandwidth dependent, crystal thickness combinations for the detailed design of compliant THG subsystems. This simplistic geometry better elucidates acceptance bandwidth limitations that are intrinsic to the crystal material.« less

Self-Phase-Matched Second-Harmonic and White-Light Generation in a Biaxial Zinc Tungstate Single Crystal

PubMed Central

Osewski, Pawel; Belardini, Alessandro; Petronijevic, Emilija; Centini, Marco; Leahu, Grigore; Diduszko, Ryszard; Pawlak, Dorota A.; Sibilia, Concita

2017-01-01

Second-order nonlinear optical materials are used to generate new frequencies by exploiting second-harmonic generation (SHG), a phenomenon where a nonlinear material generates light at double the optical frequency of the input beam. Maximum SHG is achieved when the pump and the generated waves are in phase, for example through birefringence in uniaxial crystals. However, applying these materials usually requires a complicated cutting procedure to yield a crystal with a particular orientation. Here we demonstrate the first example of phase matching under the normal incidence of SHG in a biaxial monoclinic single crystal of zinc tungstate. The crystal was grown by the micro-pulling-down method with the (102) plane perpendicular to the growth direction. Additionally, at the same time white light was generated as a result of stimulated Raman scattering and multiphoton luminescence induced by higher-order effects such as three-photon luminescence enhanced by cascaded third-harmonic generation. The annealed crystal offers SHG intensities approximately four times larger than the as grown one; optimized growth and annealing conditions may lead to much higher SHG intensities. PMID:28338074

Self-Phase-Matched Second-Harmonic and White-Light Generation in a Biaxial Zinc Tungstate Single Crystal

NASA Astrophysics Data System (ADS)

Osewski, Pawel; Belardini, Alessandro; Petronijevic, Emilija; Centini, Marco; Leahu, Grigore; Diduszko, Ryszard; Pawlak, Dorota A.; Sibilia, Concita

2017-03-01

Second-order nonlinear optical materials are used to generate new frequencies by exploiting second-harmonic generation (SHG), a phenomenon where a nonlinear material generates light at double the optical frequency of the input beam. Maximum SHG is achieved when the pump and the generated waves are in phase, for example through birefringence in uniaxial crystals. However, applying these materials usually requires a complicated cutting procedure to yield a crystal with a particular orientation. Here we demonstrate the first example of phase matching under the normal incidence of SHG in a biaxial monoclinic single crystal of zinc tungstate. The crystal was grown by the micro-pulling-down method with the (102) plane perpendicular to the growth direction. Additionally, at the same time white light was generated as a result of stimulated Raman scattering and multiphoton luminescence induced by higher-order effects such as three-photon luminescence enhanced by cascaded third-harmonic generation. The annealed crystal offers SHG intensities approximately four times larger than the as grown one; optimized growth and annealing conditions may lead to much higher SHG intensities.

3D second harmonic generation imaging tomography by multi-view excitation

PubMed Central

Campbell, Kirby R.; Wen, Bruce; Shelton, Emily M.; Swader, Robert; Cox, Benjamin L.; Eliceiri, Kevin; Campagnola, Paul J.

2018-01-01

Biological tissues have complex 3D collagen fiber architecture that cannot be fully visualized by conventional second harmonic generation (SHG) microscopy due to electric dipole considerations. We have developed a multi-view SHG imaging platform that successfully visualizes all orientations of collagen fibers. This is achieved by rotating tissues relative to the excitation laser plane of incidence, where the complete fibrillar structure is then visualized following registration and reconstruction. We evaluated high frequency and Gaussian weighted fusion reconstruction algorithms, and found the former approach performs better in terms of the resulting resolution. The new approach is a first step toward SHG tomography. PMID:29541654

Efficient third harmonic generation of a CW-fibered 1.5 µm laser diode

NASA Astrophysics Data System (ADS)

Philippe, Charles; Chea, Erick; Nishida, Yoshiki; du Burck, Frédéric; Acef, Ouali

2016-10-01

We report on frequency tripling of CW-Telecom laser diode using two cascaded PPLN ridge nonlinear crystals, both used in single-pass configuration. All optical components used for this development are fibered, leading to a very compact and easy to use optical setup. We have generated up to 290 mW optical power in the green range, from 800 mW only of infrared power around 1.54 µm. This result corresponds to an optical conversion efficiency P 3 ω / P ω > 36 %. To our knowledge, this is best value ever demonstrated up today for a CW-third harmonic generation in single-pass configuration. This frequency tripling experimental setup was tested over more than 2 years of continuous operation, without any interruption. The compactness and the reliability of our device make it very suitable as a transportable optical oscillator. In particular, it paves the way for embedded applications thanks to the high level of long-term stability of the optical alignments.

Methods, systems and apparatus for controlling third harmonic voltage when operating a multi-space machine in an overmodulation region

DOEpatents

Perisic, Milun; Kinoshita, Michael H; Ranson, Ray M; Gallegos-Lopez, Gabriel

2014-06-03

Methods, system and apparatus are provided for controlling third harmonic voltages when operating a multi-phase machine in an overmodulation region. The multi-phase machine can be, for example, a five-phase machine in a vector controlled motor drive system that includes a five-phase PWM controlled inverter module that drives the five-phase machine. Techniques for overmodulating a reference voltage vector are provided. For example, when the reference voltage vector is determined to be within the overmodulation region, an angle of the reference voltage vector can be modified to generate a reference voltage overmodulation control angle, and a magnitude of the reference voltage vector can be modified, based on the reference voltage overmodulation control angle, to generate a modified magnitude of the reference voltage vector. By modifying the reference voltage vector, voltage command signals that control a five-phase inverter module can be optimized to increase output voltages generated by the five-phase inverter module.

Continuum generation in ultra high numerical aperture fiber with application to multiphoton microscopy

NASA Astrophysics Data System (ADS)

Sayler, Nicholas

Nonlinear microscopy benefits from broadband laser sources, enabling efficient excitation of an array of fluorophores, for example. This work demonstrates broadening of a narrow band input pulse (6 nm to 40 nm) centered at 1040 nm with excellent shot-to-shot stability. In a preliminary demonstration, multiphoton imaging with pulses from the fiber is performed. In particular second harmonic imaging of corn starch is performed.

Efficient Second-Harmonic Generation in Nanocrystalline Silicon Nanoparticles.

PubMed

Makarov, Sergey V; Petrov, Mihail I; Zywietz, Urs; Milichko, Valentin; Zuev, Dmitry; Lopanitsyna, Natalia; Kuksin, Alexey; Mukhin, Ivan; Zograf, George; Ubyivovk, Evgeniy; Smirnova, Daria A; Starikov, Sergey; Chichkov, Boris N; Kivshar, Yuri S

2017-05-10

Recent trends to employ high-index dielectric particles in nanophotonics are motivated by their reduced dissipative losses and large resonant enhancement of nonlinear effects at the nanoscale. Because silicon is a centrosymmetric material, the studies of nonlinear optical properties of silicon nanoparticles have been targeting primarily the third-harmonic generation effects. Here we demonstrate, both experimentally and theoretically, that resonantly excited nanocrystalline silicon nanoparticles fabricated by an optimized laser printing technique can exhibit strong second-harmonic generation (SHG) effects. We attribute an unexpectedly high yield of the nonlinear conversion to a nanocrystalline structure of nanoparticles supporting the Mie resonances. The demonstrated efficient SHG at green light from a single silicon nanoparticle is 2 orders of magnitude higher than that from unstructured silicon films. This efficiency is significantly higher than that of many plasmonic nanostructures and small silicon nanoparticles in the visible range, and it can be useful for a design of nonlinear nanoantennas and silicon-based integrated light sources.

Second harmonic generation microscopy differentiates collagen type I and type III in COPD

NASA Astrophysics Data System (ADS)

Suzuki, Masaru; Kayra, Damian; Elliott, W. Mark; Hogg, James C.; Abraham, Thomas

2012-03-01

The structural remodeling of extracellular matrix proteins in peripheral lung region is an important feature in chronic obstructive pulmonary disease (COPD). Multiphoton microscopy is capable of inducing specific second harmonic generation (SHG) signal from non-centrosymmetric structural proteins such as fibrillar collagens. In this study, SHG microscopy was used to examine structural remodeling of the fibrillar collagens in human lungs undergoing emphysematous destruction (n=2). The SHG signals originating from these diseased lung thin sections from base to apex (n=16) were captured simultaneously in both forward and backward directions. We found that the SHG images detected in the forward direction showed well-developed and well-structured thick collagen fibers while the SHG images detected in the backward direction showed striking different morphological features which included the diffused pattern of forward detected structures plus other forms of collagen structures. Comparison of these images with the wellestablished immunohistochemical staining indicated that the structures detected in the forward direction are primarily the thick collagen type I fibers and the structures identified in the backward direction are diffusive structures of forward detected collagen type I plus collagen type III. In conclusion, we here demonstrate the feasibility of SHG microscopy in differentiating fibrillar collagen subtypes and understanding their remodeling in diseased lung tissues.

Extracting morphologies from third harmonic generation images of structurally normal human brain tissue.

PubMed

Zhang, Zhiqing; Kuzmin, Nikolay V; Groot, Marie Louise; de Munck, Jan C

2017-06-01

The morphologies contained in 3D third harmonic generation (THG) images of human brain tissue can report on the pathological state of the tissue. However, the complexity of THG brain images makes the usage of modern image processing tools, especially those of image filtering, segmentation and validation, to extract this information challenging. We developed a salient edge-enhancing model of anisotropic diffusion for image filtering, based on higher order statistics. We split the intrinsic 3-phase segmentation problem into two 2-phase segmentation problems, each of which we solved with a dedicated model, active contour weighted by prior extreme. We applied the novel proposed algorithms to THG images of structurally normal ex-vivo human brain tissue, revealing key tissue components-brain cells, microvessels and neuropil, enabling statistical characterization of these components. Comprehensive comparison to manually delineated ground truth validated the proposed algorithms. Quantitative comparison to second harmonic generation/auto-fluorescence images, acquired simultaneously from the same tissue area, confirmed the correctness of the main THG features detected. The software and test datasets are available from the authors. z.zhang@vu.nl. Supplementary data are available at Bioinformatics online. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Mid-infrared-to-mid-ultraviolet supercontinuum enhanced by third-to-fifteenth odd harmonics.

PubMed

Mitrofanov, A V; Voronin, A A; Mitryukovskiy, S I; Sidorov-Biryukov, D A; Pugžlys, A; Andriukaitis, G; Flöry, T; Stepanov, E A; Fedotov, A B; Baltuška, A; Zheltikov, A M

2015-05-01

A high-energy supercontinuum spanning 4.7 octaves, from 250 to 6500 nm, is generated using a 0.3-TW, 3.9-μm output of a mid-infrared optical parametric chirped-pulse amplifier as a driver inducing a laser filament in the air. The high-frequency wing of the supercontinuum spectrum is enhanced by odd-order optical harmonics of the mid-infrared driver. Optical harmonics up to the 15th order are observed in supercontinuum spectra as overlapping, yet well-resolved peaks broadened, as verified by numerical modeling, due to spatially nonuniform ionization-induced blue shift.

Compact double-bunch x-ray free electron lasers for fresh bunch self-seeding and harmonic lasing

DOE PAGES

Emma, C.; Feng, Y.; Nguyen, D. C.; ...

2017-03-03

This study presents a novel method to improve the longitudinal coherence, efficiency and maximum photon energy of x-ray free electron lasers (XFELs). The method is equivalent to having two separate concatenated XFELs. The first uses one bunch of electrons to reach the saturation regime, generating a high power self-amplified spontaneous emission x-ray pulse at the fundamental and third harmonic. The x-ray pulse is filtered through an attenuator/monochromator and seeds a different electron bunch in the second FEL, using the fundamental and/or third harmonic as an input signal. In our method we combine the two XFELs operating with two bunches, separatedmore » by one or more rf cycles, in the same linear accelerator. We discuss the advantages and applications of the proposed system for present and future XFELs.« less

Third-harmonic generation in tunable nonlinear hyperbolic metamaterial

NASA Astrophysics Data System (ADS)

Wicharn, Surawut; Buranasiri, Prathan

2018-03-01

In this research, a third-harmonic generation (THG) in a tunable nonlinear hyperbolic metamaterial (TNHM) has been investigated numerically. The TNHM is consisted of periodically arranging of multilayered graphene layers system for controlled optical properties purpose, and ordinary nonlinear dielectric layer. The possibility of TNHM permittivity dispersion controlled by number of graphene layers and external bias voltage to graphene layers was satisfied, then the structure has created the nearly perfect phase-matching scheme based on epsilon-near-zero (ENZ) behavior of the nonlinear medium. Finally, the optimal designed TNHM structure with sufficient bias voltage have given the forwardand backward-direction TH pulses, which the backward-forward TH intensity ratio is closely unity. The THG conversion efficiencies have been maximized after increasing the pumping level to 800 MW/cm2 . From this study, the optimal designed TNHM can be applied as a bi-directional nonlinear frequency converters in nanophotonic systems.

Optical third harmonic generation in the magnetic semiconductor EuSe

NASA Astrophysics Data System (ADS)

Lafrentz, M.; Brunne, D.; Kaminski, B.; Pavlov, V. V.; Pisarev, R. V.; Henriques, A. B.; Yakovlev, D. R.; Springholz, G.; Bauer, G.; Bayer, M.

2012-01-01

Third harmonic generation (THG) has been studied in europium selenide EuSe in the vicinity of the band gap at 2.1-2.6 eV and at higher energies up to 3.7 eV. EuSe is a magnetic semiconductor crystalizing in centrosymmetric structure of rock-salt type with the point group m3m. For this symmetry the crystallographic and magnetic-field-induced THG nonlinearities are allowed in the electric-dipole approximation. Using temperature, magnetic field, and rotational anisotropy measurements, the crystallographic and magnetic-field-induced contributions to THG were unambiguously separated. Strong resonant magnetic-field-induced THG signals were measured at energies in the range of 2.1-2.6 eV and 3.1-3.6 eV for which we assign to transitions from 4f7 to 4f65d1 bands, namely involving 5d(t2g) and 5d(eg) states.

Vibrational excitation of hydrogen molecules by two-photon absorption and third-harmonic generation

NASA Astrophysics Data System (ADS)

Miyamoto, Yuki; Hara, Hideaki; Hiraki, Takahiro; Masuda, Takahiko; Sasao, Noboru; Uetake, Satoshi; Yoshimi, Akihiro; Yoshimura, Koji; Yoshimura, Motohiko

2018-01-01

We report the coherent excitation of the vibrational state of hydrogen molecules by two-photon absorption and the resultant third-harmonic generation (THG). Parahydrogen molecules cooled by liquid nitrogen are irradiated by mid-infrared nanosecond pulses at 4.8 μm with a nearly Fourier-transform-limited linewidth. The first excited vibrational state of parahydrogen is populated by two-photon absorption of the mid-infrared photons. Because of the narrow linewidth of the mid-infrared pulses, coherence between the ground and excited states is sufficient to induce higher-order processes. Near-infrared photons from the THG are observed at 1.6 μm. The dependence of the intensity of the near-infrared radiation on mid-infrared pulse energy, target pressure, and cell length is determined. We used a simple formula for THG with consideration of realistic experimental conditions to explain the observed results.

Role of phase matching in pulsed second-harmonic generation: Walk-off and phase-locked twin pulses in negative-index media

NASA Astrophysics Data System (ADS)

Roppo, Vito; Centini, Marco; Sibilia, Concita; Bertolotti, Mario; de Ceglia, Domenico; Scalora, Michael; Akozbek, Neset; Bloemer, Mark J.; Haus, Joseph W.; Kosareva, Olga G.; Kandidov, Valery P.

2007-09-01

The present investigation is concerned with the study of pulsed second-harmonic generation under conditions of phase and group velocity mismatch, and generally low conversion efficiencies and pump intensities. In positive-index, nonmetallic materials, we generally find qualitative agreement with previous reports regarding the presence of a double-peaked second harmonic signal, which comprises a pulse that walks off and propagates at the nominal group velocity one expects at the second-harmonic frequency, and a second pulse that is “captured” and propagates under the pump pulse. We find that the origin of the double-peaked structure resides in a phase-locking mechanism that characterizes not only second-harmonic generation, but also χ(3) processes and third-harmonic generation. The phase-locking mechanism that we describe occurs for arbitrarily small pump intensities, and so it is not a soliton effect, which usually relies on a threshold mechanism, although multicolor solitons display similar phase locking characteristics. Thus, in second harmonic generation a phase-matched component is always generated, even under conditions of material phase mismatch: This component is anomalous, because the material does not allow energy exchange between the pump and the second-harmonic beam. On the other hand, if the material is phase matched, phase locking and phase matching are indistinguishable, and the conversion process becomes efficient. We also report a similar phase-locking phenomenon in negative index materials. A spectral analysis of the pump and the generated signals reveals that the phase-locking phenomenon causes the forward moving, phase-locked second-harmonic pulse to experience the same negative index as the pump pulse, even though the index of refraction at the second-harmonic frequency is positive. Our analysis further shows that the reflected second-harmonic pulse generated at the interface and the forward-moving, phase-locked pulse appear to be part of the same pulse initially generated at the surface, part of which is immediately back-reflected, while the rest becomes trapped and dragged along by the pump pulse. These pulses thus constitute twin pulses generated at the interface, having the same negative wave vector, but propagating in opposite directions. Almost any break of the longitudinal symmetry, even an exceedingly small χ(2) discontinuity, releases the trapped pulse which then propagates in the backward direction. These dynamics are indicative of very rich and intricate interactions that characterize ultrashort pulse propagation phenomena.

Role of phase matching in pulsed second-harmonic generation: Walk-off and phase-locked twin pulses in negative-index media

DOE Office of Scientific and Technical Information (OSTI.GOV)

Roppo, Vito; Centini, Marco; Sibilia, Concita

The present investigation is concerned with the study of pulsed second-harmonic generation under conditions of phase and group velocity mismatch, and generally low conversion efficiencies and pump intensities. In positive-index, nonmetallic materials, we generally find qualitative agreement with previous reports regarding the presence of a double-peaked second harmonic signal, which comprises a pulse that walks off and propagates at the nominal group velocity one expects at the second-harmonic frequency, and a second pulse that is 'captured' and propagates under the pump pulse. We find that the origin of the double-peaked structure resides in a phase-locking mechanism that characterizes not onlymore » second-harmonic generation, but also {chi}{sup (3)} processes and third-harmonic generation. The phase-locking mechanism that we describe occurs for arbitrarily small pump intensities, and so it is not a soliton effect, which usually relies on a threshold mechanism, although multicolor solitons display similar phase locking characteristics. Thus, in second harmonic generation a phase-matched component is always generated, even under conditions of material phase mismatch: This component is anomalous, because the material does not allow energy exchange between the pump and the second-harmonic beam. On the other hand, if the material is phase matched, phase locking and phase matching are indistinguishable, and the conversion process becomes efficient. We also report a similar phase-locking phenomenon in negative index materials. A spectral analysis of the pump and the generated signals reveals that the phase-locking phenomenon causes the forward moving, phase-locked second-harmonic pulse to experience the same negative index as the pump pulse, even though the index of refraction at the second-harmonic frequency is positive. Our analysis further shows that the reflected second-harmonic pulse generated at the interface and the forward-moving, phase-locked pulse appear to be part of the same pulse initially generated at the surface, part of which is immediately back-reflected, while the rest becomes trapped and dragged along by the pump pulse. These pulses thus constitute twin pulses generated at the interface, having the same negative wave vector, but propagating in opposite directions. Almost any break of the longitudinal symmetry, even an exceedingly small {chi}{sup (2)} discontinuity, releases the trapped pulse which then propagates in the backward direction. These dynamics are indicative of very rich and intricate interactions that characterize ultrashort pulse propagation phenomena.« less

Effect of composition and temperature on the second harmonic generation in silver phosphate glasses

NASA Astrophysics Data System (ADS)

Konidakis, I.; Psilodimitrakopoulos, S.; Kosma, K.; Lemonis, A.; Stratakis, E.

2018-01-01

We herein employ nonlinear laser imaging microscopy to explicitly study the dynamics of second harmonic generation (SHG) in silver iodide phosphate glasses. While glasses of this family have gained extensive scientific attention over the years due to their superior conducting properties, considerably less attention has been paid to their unique nonlinear optical characteristics. In the present study, firstly, it is demonstrated that SHG signal intensity is enhanced upon increasing silver content due to the random formation of silver microstructures within the glass network. Secondly, the SHG temperature dynamics were explored near the glass transition temperature (Tg) regime, where significant glass relaxation phenomena occur. It is found that heating towards the Tg improves the SHG efficiency, whereas above Tg, the capacity of glasses to generate second harmonic radiation is drastically suppressed. The novel findings of this work are considered important in terms of the potential employment of these glasses for the realization of advanced photonic applications like optical-switches and wavelength conversion devices.

Investigation of the effect of hydration on dermal collagen in ex vivo human skin tissue using second harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Samatham, Ravikant; Wang, Nicholas K.; Jacques, Steven L.

2016-02-01

Effect of hydration on the dermal collagen structure in human skin was investigated using second harmonic generation microscopy. Dog ears from the Mohs micrographic surgery department were procured for the study. Skin samples with subject aged between 58-90 years old were used in the study. Three dimensional Multiphoton (Two-photon and backward SHG) control data was acquired from the skin samples. After the control measurement, the skin tissue was either soaked in deionized water for 2 hours (Hydration) or kept at room temperature for 2 hours (Desiccation), and SHG data was acquired. The data was normalized for changes in laser power and detector gain. The collagen signal per unit volume from the dermis was calculated. The desiccated skin tissue gave higher backward SHG compared to respective control tissue, while hydration sample gave a lower backward SHG. The collagen signal decreased with increase in hydration of the dermal collagen. Hydration affected the packing of the collagen fibrils causing a change in the backward SHG signal. In this study, the use of multiphoton microscopy to study the effect of hydration on dermal structure was demonstrated in ex vivo tissue.

Motion-artifact-robust, polarization-resolved second-harmonic-generation microscopy based on rapid polarization switching with electro-optic Pockells cell and its application to in vivo visualization of collagen fiber orientation in human facial skin

PubMed Central

Tanaka, Yuji; Hase, Eiji; Fukushima, Shuichiro; Ogura, Yuki; Yamashita, Toyonobu; Hirao, Tetsuji; Araki, Tsutomu; Yasui, Takeshi

2014-01-01

Polarization-resolved second-harmonic-generation (PR-SHG) microscopy is a powerful tool for investigating collagen fiber orientation quantitatively with low invasiveness. However, the waiting time for the mechanical polarization rotation makes it too sensitive to motion artifacts and hence has hampered its use in various applications in vivo. In the work described in this article, we constructed a motion-artifact-robust, PR-SHG microscope based on rapid polarization switching at every pixel with an electro-optic Pockells cell (PC) in synchronization with step-wise raster scanning of the focus spot and alternate data acquisition of a vertical-polarization-resolved SHG signal and a horizontal-polarization-resolved one. The constructed PC-based PR-SHG microscope enabled us to visualize orientation mapping of dermal collagen fiber in human facial skin in vivo without the influence of motion artifacts. Furthermore, it implied the location and/or age dependence of the collagen fiber orientation in human facial skin. The robustness to motion artifacts in the collagen orientation measurement will expand the application scope of SHG microscopy in dermatology and collagen-related fields. PMID:24761292

In vivo multiphoton and second harmonic generation microscopy of epithelial carcinogenesis

NASA Astrophysics Data System (ADS)

Vargas, Gracie; Shilagard, Tuya; Sun, Ju; Motamedi, Massoud

2006-02-01

Multiphoton microscopy and second harmonic generation microscopy were used to image epithelial changes in a hamster model for oral malignant transformation. In vivo imaging was performed to characterize morphometric alterations in normal and precancerous regions. Morphometric measurements such as cell nucleus area and epithelial thicknesses obtained from MPM-SHGM were in excellent agreement with histology obtained after in vivo imaging. MPM-SHGM was highly sensitive to spectroscopic and architectural alterations throughout carcinogenesis, showing statistically significant changes in morphology. MPM revealed hyperkeratosis, nuclear enlargement/crowding in dysplasia, and immune cell infiltration. SHGM revealed alterations in submucosal architecture, with a decrease in SHG density evident during early stages of precancer. By combining MPM with SHGM, the basement membrane could be identified in normal, hyperplasia, and dysplasia samples and in some cases of early invasion. The combined technique of MPM-SHGM has the potential to serve as an adjunct to biopsy for assessing precancerous changes and will be investigated further for that purpose. Additionally, the method can provide spatiotemporal assessment of early neoplastic changes in order to elucidate the stages of transformation in vivo and could be used to assess therapeutic efficacy of agents being tested for the treatment of epithelial precancers/cancer.

Nonlinear optical microscopy: use of second harmonic generation and two-photon microscopy for automated quantitative liver fibrosis studies.

PubMed

Sun, Wanxin; Chang, Shi; Tai, Dean C S; Tan, Nancy; Xiao, Guangfa; Tang, Huihuan; Yu, Hanry

2008-01-01

Liver fibrosis is associated with an abnormal increase in an extracellular matrix in chronic liver diseases. Quantitative characterization of fibrillar collagen in intact tissue is essential for both fibrosis studies and clinical applications. Commonly used methods, histological staining followed by either semiquantitative or computerized image analysis, have limited sensitivity, accuracy, and operator-dependent variations. The fibrillar collagen in sinusoids of normal livers could be observed through second-harmonic generation (SHG) microscopy. The two-photon excited fluorescence (TPEF) images, recorded simultaneously with SHG, clearly revealed the hepatocyte morphology. We have systematically optimized the parameters for the quantitative SHG/TPEF imaging of liver tissue and developed fully automated image analysis algorithms to extract the information of collagen changes and cell necrosis. Subtle changes in the distribution and amount of collagen and cell morphology are quantitatively characterized in SHG/TPEF images. By comparing to traditional staining, such as Masson's trichrome and Sirius red, SHG/TPEF is a sensitive quantitative tool for automated collagen characterization in liver tissue. Our system allows for enhanced detection and quantification of sinusoidal collagen fibers in fibrosis research and clinical diagnostics.

Non-Linear Optical Microscopy Sheds Light on Cardiovascular Disease

PubMed Central

Caorsi, Valentina; Toepfer, Christopher; Sikkel, Markus B.; Lyon, Alexander R.; MacLeod, Ken; Ferenczi, Mike A.

2013-01-01

Many cardiac diseases have been associated with increased fibrosis and changes in the organization of fibrillar collagen. The degree of fibrosis is routinely analyzed with invasive histological and immunohistochemical methods, giving a limited and qualitative understanding of the tissue's morphological adaptation to disease. Our aim is to quantitatively evaluate the increase in fibrosis by three-dimensional imaging of the collagen network in the myocardium using the non-linear optical microscopy techniques Two-Photon Excitation microscopy (TPE) and Second Harmonic signal Generation (SHG). No sample staining is needed because numerous endogenous fluorophores are excited by a two-photon mechanism and highly non-centrosymmetric structures such as collagen generate strong second harmonic signals. We propose for the first time a 3D quantitative analysis to carefully evaluate the increased fibrosis in tissue from a rat model of heart failure post myocardial infarction. We show how to measure changes in fibrosis from the backward SHG (BSHG) alone, as only backward-propagating SHG is accessible for true in vivo applications. A 5-fold increase in collagen I fibrosis is detected in the remote surviving myocardium measured 20 weeks after infarction. The spatial distribution is also shown to change markedly, providing insight into the morphology of disease progression. PMID:23409139

Realization of a mW-level 10.7-eV (λ = 115.6 nm) laser by cascaded third harmonic generation of a Yb:fiber CPA laser at 1-MHz.

PubMed

Zhao, Zhigang; Kobayashi, Yohei

2017-06-12

We demonstrate a 10.7-eV (λ = 115.6 nm) laser with mW levels of average power and a 1-MHz repetition rate, which was driven by the third harmonic radiation (THG), at 347 nm, of an Yb:fiber chirped pulse amplifier (CPA) laser. The 347 nm ultraviolet radiation was obtained by frequency conversion of the high power output of a 1-MHz Yb:fiber CPA, using beta barium borate (BBO) nonlinear crystals. The frequency converted output was focused down into a gas cell filled with a mixture of Ar and Xe, and was subjected to a second THG frequency conversion. The generated 10.7-eV laser was separated from the fundamental beam using a LiF prism and no further separation from other harmonic waves was required. The highest measured output power was ~80 μW, which corresponded to an average power of ~1.25 mW inside the gas cell when the transmission coefficients of the LiF optics were taken into account. The corresponding conversion efficiency from 347 nm down to 115.6 nm was ~2.5 × 10 -4 .

Non-label bioimaging utilizing scattering lights

NASA Astrophysics Data System (ADS)

Watanabe, Tomonobu M.; Ichimura, Taro; Fujita, Hideaki

2017-04-01

Optical microscopy is an indispensable tool for medical and life sciences. Especially, the microscopes utilized with scattering light offer a detailed internal observation of living specimens in real time because of their non-labeling and non-invasive capability. We here focus on two kinds of scattering lights, Raman scattering light and second harmonic generation light. Raman scattering light includes the information of all the molecular vibration modes of the molecules, and can be used to distinguish types and/or state of cell. Second harmonic generation light is derived from electric polarity of proteins in the specimen, and enables to detect their structural change. In this conference, we would like to introduce our challenges to extract biological information from those scattering lights.

Two-photon spectral fluorescence lifetime and second-harmonic generation imaging of the porcine cornea with a 12-femtosecond laser microscope

NASA Astrophysics Data System (ADS)

Batista, Ana; Breunig, Hans Georg; Uchugonova, Aisada; Morgado, António Miguel; König, Karsten

2016-03-01

Five dimensional microscopy with a 12-fs laser scanning microscope based on spectrally resolved two-photon autofluorescence lifetime and second-harmonic generation (SHG) imaging was used to characterize all layers of the porcine cornea. This setup allowed the simultaneous excitation of both metabolic cofactors, NAD(P)H and flavins, and their discrimination based on their spectral emission properties and fluorescence decay characteristics. Furthermore, the architecture of the stromal collagen fibrils was assessed by SHG imaging in both forward and backward directions. Information on the metabolic state and the tissue architecture of the porcine cornea were obtained with subcellular resolution, and high temporal and spectral resolutions.

Two-photon spectral fluorescence lifetime and second-harmonic generation imaging of the porcine cornea with a 12-femtosecond laser microscope.

PubMed

Batista, Ana; Breunig, Hans Georg; Uchugonova, Aisada; Morgado, António Miguel; König, Karsten

2016-03-01

Five dimensional microscopy with a 12-fs laser scanning microscope based on spectrally resolved two-photon autofluorescence lifetime and second-harmonic generation (SHG) imaging was used to characterize all layers of the porcine cornea. This setup allowed the simultaneous excitation of both metabolic cofactors, NAD(P)H and flavins, and their discrimination based on their spectral emission properties and fluorescence decay characteristics. Furthermore, the architecture of the stromal collagen fibrils was assessed by SHG imaging in both forward and backward directions. Information on the metabolic state and the tissue architecture of the porcine cornea were obtained with subcellular resolution, and high temporal and spectral resolutions.

Determination of three-dimensional molecular orientation of type-I collagen by circularly-polarized second harmonic generation imaging

NASA Astrophysics Data System (ADS)

Zhuo, Guan-Yu; Hung, Wei-Han; Kao, Fu-Jen

2017-04-01

The content of collagen is up to 30% existing in mammals. It supports the main component of connective tissues such as skin, ligament, and cartilage. Among various types of collagen, type-I collagen is of the most abundance and has been broadly studied due to the importance in bioscience. Second harmonic generation (SHG) microscopy is an effective tool used to study the collagen organization without labeling. In this study, we used circular polarization instead of linear polarization to retrieve three-dimensional (3D) molecular orientation of type-I collagen with only two cross polarized SHG images without acquiring an image stack of varying polarization.

Bioinspired second harmonic generation

NASA Astrophysics Data System (ADS)

Sonay, Ali Y.; Pantazis, Periklis

2017-07-01

Second harmonic generation (SHG) is a microscopic technique applicable to a broad spectrum of biological and medical imaging due to its excellent photostability, high signal-to-noise ratio (SNR) and narrow emission profile. Current SHG microscopy techniques rely on two main contrast modalities. These are endogenous SHG generated by tissue structures, which is clinically relevant but cannot be targeted to another location, or SHG nanoprobes, inorganic nanocrystals that can be directed to proteins and cells of interest, but cannot be applied for clinical imaging due to their chemical composition. Here we analyzed SHG signal generated by large-scale peptide assemblies. Our results show the sequence of peptides play an important role on both the morphology and SHG signal of the peptide assemblies. Changing peptide sequence allows confinement of large number of peptides to smaller voxels, generating intense SHG signal. With miniaturization of these peptides and their proper functionalization strategies, such bioinspired nanoparticles would emerge as valuable tools for clinical imaging.

Photon energy conversion by near-zero permittivity nonlinear materials

DOE Office of Scientific and Technical Information (OSTI.GOV)

Luk, Ting S.; Sinclair, Michael B.; Campione, Salvatore

Efficient harmonic light generation can be achieved with ultrathin films by coupling an incident pump wave to an epsilon-near-zero (ENZ) mode of the thin film. As an example, efficient third harmonic generation from an indium tin oxide nanofilm (.lamda./42 thick) on a glass substrate for a pump wavelength of 1.4 .mu.m was demonstrated. A conversion efficiency of 3.3.times.10.sup.-6 was achieved by exploiting the field enhancement properties of the ENZ mode with an enhancement factor of 200. This nanoscale frequency conversion method is applicable to other plasmonic materials and reststrahlen materials in proximity of the longitudinal optical phonon frequencies.

Cardiovascular Imaging Using Two-Photon Microscopy

PubMed Central

Scherschel, John A.; Rubart, Michael

2008-01-01

Two-photon excitation microscopy has become the standard technique for high resolution deep tissue and intravital imaging. It provides intrinsic three-dimensional resolution in combination with increased penetration depth compared to single-photon confocal microscopy. This article will describe the basic physical principles of two-photon excitation and will review its multiple applications to cardiovascular imaging, including second harmonic generation and fluorescence laser scanning microscopy. In particular, the capability and limitations of multiphoton microscopy to assess functional heterogeneity on a cellular scale deep within intact, Langendorff-perfused hearts are demonstrated. It will also discuss the use of two-photon excitation-induced release of caged compounds for the study of intracellular calcium signaling and intercellular dye transfer. PMID:18986603

Optical bistability and second-harmonic generation in thin film coupled cavity photonic crystal structures

NASA Astrophysics Data System (ADS)

Diao, Liyong

This thesis deals with design, fabrication and modeling of bistable and multi-stable switching dynamics and second-harmonic generation in two groups of thin film coupled cavity photonic crystal structures. The first component studies optical bistability and multistability in such structures. Optical bistability and multistability are modelled by a nonlinear transfer matrix method. The second component is focused on the modelling and experimental measurement of second-harmonic generation in such structures. It is found that coupled cavity structures can reduce the threshold and index change for bistable operation, but single cavity structures can do the same. However, there is a clear advantage in using coupled cavity structures for multistability in that the threshold for multistability can be reduced. Second-harmonic generation is enhanced by field localization due to the resonant effect at the fundamental wavelength in single and coupled cavity structures by simulated and measured results. The work in this thesis makes three significant contributions. First, in the successful fabrication of thin film coupled cavity structures, the simulated linear transmissions of such structures match those of the fabricated structures almost exactly. Second, the newly defined figure of merit at the maximum transmission point on the bistable curve can be used to compare the material damage tolerance to any other Kerr effect nonlinear gate. Third, the simulated second-harmonic generation agrees excellently with experimental results. More generally optical thin film fabrication has commercial applications in many industry sections, such as electronics, opto-electronics, optical coating, solar cell and MEMS.

Third order harmonic imaging for biological tissues using three phase-coded pulses.

PubMed

Ma, Qingyu; Gong, Xiufen; Zhang, Dong

2006-12-22

Compared to the fundamental and the second harmonic imaging, the third harmonic imaging shows significant improvements in image quality due to the better resolution, but it is degraded by the lower sound pressure and signal-to-noise ratio (SNR). In this study, a phase-coded pulse technique is proposed to selectively enhance the sound pressure of the third harmonic by 9.5 dB whereas the fundamental and the second harmonic components are efficiently suppressed and SNR is also increased by 4.7 dB. Based on the solution of the KZK nonlinear equation, the axial and lateral beam profiles of harmonics radiated from a planar piston transducer were theoretically simulated and experimentally examined. Finally, the third harmonic images using this technique were performed for several biological tissues and compared with the images obtained by the fundamental and the second harmonic imaging. Results demonstrate that the phase-coded pulse technique yields a dramatically cleaner and sharper contrast image.

Second Harmonic Generation Mediated by Aligned Water in Starch Granules.

PubMed

Cisek, Richard; Tokarz, Danielle; Krouglov, Serguei; Steup, Martin; Emes, Michael J; Tetlow, Ian J; Barzda, Virginijus

2014-12-26

The origin of second harmonic generation (SHG) in starch granules was investigated using ab initio quantum mechanical modeling and experimentally examined using polarization-in, polarization-out (PIPO) second harmonic generation microscopy. Ab initio calculations revealed that the largest contribution to the SHG signal from A- and B-type allomorphs of starch originates from the anisotropic organization of hydroxide and hydrogen bonds mediated by aligned water found in the polymers. The hypothesis was experimentally tested by imaging maize starch granules under various hydration and heat treatment conditions that alter the hydrogen bond network. The highest SHG intensity was found in fully hydrated starch granules, and heat treatment diminished the SHG intensity. The PIPO SHG imaging showed that dried starch granules have a much higher nonlinear optical susceptibility component ratio than fully hydrated granules. In contrast, deuterated starch granules showed a smaller susceptibility component ratio demonstrating that SHG is highly sensitive to the organization of the hydroxyl and hydrogen bond network. The polarization SHG imaging results of potato starch granules, representing starch allomorph B, were compared to those of maize starch granules representing allomorph A. The results showed that the amount of aligned water was higher in the maize granules. Nonlinear microscopy of starch granules provides evidence that varying hydration conditions leads to significant changes in the nonlinear susceptibility ratio as well as the SHG intensity, supporting the hypothesis from ab initio calculations that the dominant contribution to SHG is due to the ordered hydroxide and hydrogen bond network.

Polarization-Modulated Second Harmonic Generation Microscopy in Collagen

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stoller, P C

Collagen is a key structural protein in the body; several pathological conditions lead to changes in collagen. Among imaging modalities that can be used in vivo, second harmonic generation (SHG) microscopy has a key advantage: it provides {approx}1 {micro}m resolution information about collagen structure as a function of depth. A new technique--polarization-modulated SHG--is presented: it permits simultaneous measurement of collagen orientation, of a lower bound on the magnitude of the second order nonlinear susceptibility tensor, and of the ratio of the two independent elements in this tensor. It is applied to characterizing SHG in collagen and to determining effects ofmore » biologically relevant changes in collagen structure. The magnitude of the second harmonic signal in two dimensional images varies with position even in structurally homogeneous tissue; this phenomenon is due to interference between second harmonic light generated by neighboring fibrils, which are randomly oriented parallel or anti-parallel to each other. Studies in which focal spot size was varied indicated that regions where fibrils are co-oriented are less than {approx}1.5 {micro}m in diameter. A quartz reference was used to determine the spot size as well as a lower limit (d{sub xxx} > 0.3 pm/V) for the magnitude of the second order nonlinear susceptibility. The ratio of the two independent tensor elements ranged between d{sub XYY}/d{sub XXX} = 0.60 and 0.75. SHG magnitude alone was not useful for identifying structural anomalies in collagenous tissue. Instead, changes in the polarization dependence of SHG were used to analyze biologically relevant perturbations in collagen structure. Changes in polarization dependence were observed in dehydrated samples, but not in highly crosslinked samples, despite significant alterations in packing structure. Complete thermal denaturation and collagenase digestion produced samples with no detectable SHG signal. Collagen orientation was measured in thin samples of several different tissues in transmission mode as well as at different depths (up to 200 {micro}m) in thick samples in reflection mode; birefringence had no effect on the measurement. These studies showed that SHG microscopy was capable of detecting pathophysiological changes in collagen structure, suggesting that this technique has potential clinical applications.« less

Using Second Harmonic Generation Microscopy to Study the Three-Dimensional Structure of Collagen and its Degradation Mechanism

NASA Astrophysics Data System (ADS)

Mega, Yair

Collagen is one of the most abundant proteins found in the human body. Its crystalline structure possesses no centrosymmetry, allowing it to emit second-harmonic waves. Second harmonic generation (SHG) microscopy utilizes the latter quality to produce high-resolution images of collagen rich tissues and therefore become a key research tool in the biomedical field. We developed a new model, intended to be used together with second harmonic generation (SHG) microscopy, to thoroughly investigate collagen-based tissues. We use our SHG model to reveal information in real time from enzymatic biochemical processes. We also present a novel method used to measure quantitatively the direction of the fibers within the tissue, from SHG images. Using this method, we were able to reconstruct an angular map of the orientation of collagen fibers from multiple sections across the entire area of a human cornea. The structure we obtained demonstrates the criss-crossing structure of the human cornea, previously suggested in the literature. In addition, we also report work on a unique step-wise three-photon fluorescence excitation discovered in melanin. This unique fluorescence mechanism was exploited to discriminate melanin on a small-size, low-cost and low laser power setup which was used as a prototype for a handheld device. The latter study is a part of a larger on-going effort in our group to explore new diagnosis methods to be used for early skin cancer screening. Finally, this work demonstrates a spectroscopy-based method to correct for blood vessel thickness effect. The method analyzes spectral shift from a molecular imaging agent and correlate the shifts to the length of the optical path in blood. The correction method described in this work is intended to be implemented on a guided catheter near infrared fluorescence (NIRF) intra-vascular imaging system. In this imaging system, this study's results will used to correct for the radial distance between the imaging tip of the catheter and fluorescing agents chemically bonded to plaques on walls of the arteries.

Two-Photon and Second Harmonic Microscopy in Clinical and Translational Cancer Research

PubMed Central

PERRY, SETH W.; BURKE, RYAN M.; BROWN, EDWARD B.

2012-01-01

Application of two-photon microscopy (TPM) to translational and clinical cancer research has burgeoned over the last several years, as several avenues of pre-clinical research have come to fruition. In this review, we focus on two forms of TPM—two-photon excitation fluorescence microscopy, and second harmonic generation microscopy—as they have been used for investigating cancer pathology in ex vivo and in vivo human tissue. We begin with discussion of two-photon theory and instrumentation particularly as applicable to cancer research, followed by an overview of some of the relevant cancer research literature in areas that include two-photon imaging of human tissue biopsies, human skin in vivo, and the rapidly developing technology of two-photon microendoscopy. We believe these and other evolving two-photon methodologies will continue to help translate cancer research from the bench to the bedside, and ultimately bring minimally invasive methods for cancer diagnosis and treatment to therapeutic reality. PMID:22258888

Nonlinear Optics and Organic Materials

DTIC Science & Technology

1989-10-01

incrementally by making small changes in the generating optical harmonics. However, deficiencies in backbone or substituents. In this way the chemist can...experimental determination of Otx.l = 4.5 X 10-32 esu. ability of polymeric molecules to generate third Key parameters extracted from the UV and visible...solubility of most active organics in negative charge at the other end, thus generating a the polymer and their tendency to segregate or migrate out

Generation of UV light by intense ultrashort laser pulses in air

NASA Astrophysics Data System (ADS)

Alexeev, Ilya; Ting, Antonio; Gordon, Daniel; Briscoe, Eldridge; Penano, Joe; Sprangle, Phillip

2004-11-01

The propagation of collimated high-peak-power ultrashort laser pulses in air has attracted considerable attention, which may have a variety of important applications including remote sensing and chemical-biological aerosols standoff detection. Sub-millimeter diameter laser filaments can develop without any focusing optics and instead solely from laser self-focusing and plasma formation in air. These filaments can produce ultraviolet radiations in the form of the 3rd harmonic of the fundamental frequency and also through spectral broadening due to self-phase modulation of the laser pulse. Using femtosecond laser pulses produced by a high power Ti:Sapphire laser (0.8 TW, 50 fs, 800 nm) we observed generation of the third harmonic radiation light in air (centered around 267 nm) by the laser filaments. Characterization of the 3rd harmonic generation with respect to the major gas components of the air will be reported. Supported by the ONR and RDECOM. I. Alexeev is NRC/NRL Post-Doc.

High-harmonic generation by field enhanced femtosecond pulses in metal-sapphire nanostructure

PubMed Central

Han, Seunghwoi; Kim, Hyunwoong; Kim, Yong Woo; Kim, Young-Jin; Kim, Seungchul; Park, In-Yong; Kim, Seung-Woo

2016-01-01

Plasmonic high-harmonic generation (HHG) drew attention as a means of producing coherent extreme ultraviolet (EUV) radiation by taking advantage of field enhancement occurring in metallic nanostructures. Here a metal-sapphire nanostructure is devised to provide a solid tip as the HHG emitter, replacing commonly used gaseous atoms. The fabricated solid tip is made of monocrystalline sapphire surrounded by a gold thin-film layer, and intended to produce EUV harmonics by the inter- and intra-band oscillations of electrons driven by the incident laser. The metal-sapphire nanostructure enhances the incident laser field by means of surface plasmon polaritons, triggering HHG directly from moderate femtosecond pulses of ∼0.1 TW cm−2 intensities. The measured EUV spectra exhibit odd-order harmonics up to ∼60 nm wavelengths without the plasma atomic lines typically seen when using gaseous atoms as the HHG emitter. This experimental outcome confirms that the plasmonic HHG approach is a promising way to realize coherent EUV sources for nano-scale near-field applications in spectroscopy, microscopy, lithography and atto-second physics. PMID:27721374

Second-harmonic generation in shear wave beams with different polarizations

NASA Astrophysics Data System (ADS)

Spratt, Kyle S.; Ilinskii, Yurii A.; Zabolotskaya, Evgenia A.; Hamilton, Mark F.

2015-10-01

A coupled pair of nonlinear parabolic equations was derived by Zabolotskaya [1] that model the transverse components of the particle motion in a collimated shear wave beam propagating in an isotropic elastic solid. Like the KZK equation, the parabolic equation for shear wave beams accounts consistently for the leading order effects of diffraction, viscosity and nonlinearity. The nonlinearity includes a cubic nonlinear term that is equivalent to that present in plane shear waves, as well as a quadratic nonlinear term that is unique to diffracting beams. The work by Wochner et al. [2] considered shear wave beams with translational polarizations (linear, circular and elliptical), wherein second-order nonlinear effects vanish and the leading order nonlinear effect is third-harmonic generation by the cubic nonlinearity. The purpose of the current work is to investigate the quadratic nonlinear term present in the parabolic equation for shear wave beams by considering second-harmonic generation in Gaussian beams as a second-order nonlinear effect using standard perturbation theory. In order for second-order nonlinear effects to be present, a broader class of source polarizations must be considered that includes not only the familiar translational polarizations, but also polarizations accounting for stretching, shearing and rotation of the source plane. It is found that the polarization of the second harmonic generated by the quadratic nonlinearity is not necessarily the same as the polarization of the source-frequency beam, and we are able to derive a general analytic solution for second-harmonic generation from a Gaussian source condition that gives explicitly the relationship between the polarization of the source-frequency beam and the polarization of the second harmonic.

Multiphoton microscopic imaging of human normal and cancerous oesophagus tissue.

PubMed

Chen, W S; Wang, Y; Liu, N R; Zhang, J X; Chen, R

2014-01-01

In this paper, microstructures of human oesophageal submucosa are evaluated using multiphoton microscopy, based on two-photon excited fluorescence and second harmonic generation. The content and distribution of collagen, elastic fibers and cancer cells in normal and cancerous submucosa layer have been distinctly obtained and briefly discussed. The variation of these components is very relevant to the pathology in oesophagus, especially in early oesophageal cancer. Our results further indicate that the multiphoton microscopy technique has the potential application in vivo in clinical diagnosis and monitoring of early oesophageal cancer. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

Strong guided mode resonant local field enhanced visible harmonic generation in an azo-polymer resonant waveguide grating.

PubMed

Lin, Jian Hung; Tseng, Chun-Yen; Lee, Ching-Ting; Young, Jeff F; Kan, Hung-Chih; Hsu, Chia Chen

2014-02-10

Guided mode resonance (GMR) enhanced second- and third-harmonic generation (SHG and THG) is demonstrated in an azo-polymer resonant waveguide grating (RWG), comprised of a poled azo-polymer layer on top of a textured SU8 substrate with a thin intervening layer of TiO2. Strong SHG and THG outputs are observed by matching either in-coming fundamental- or out-going harmonic-wavelength to the GMR wavelengths of the azo-polymer RWG. Without the azo-polymer coating, pure TiO2 RWGs, do not generate any detectable SHG using a fundamental beam peak intensity of 2 MW/cm(2). Without the textured TiO2 layer, a planar poled azo-polymer layer results in 3650 times less SHG than the full nonlinear RWG structure under identical excitation conditions. Rigorous coupled-wave analysis calculations confirm that this enhancement of the nonlinear conversion is due to strong local electric fields that are generated at the interfaces of the TiO2 and azo-polymer layers when the RWG is excited at resonant wavelengths associated with both SHG and THG conversion processes.

Odd harmonics-enhanced supercontinuum in bulk solid-state dielectric medium.

PubMed

Garejev, N; Jukna, V; Tamošauskas, G; Veličkė, M; Šuminas, R; Couairon, A; Dubietis, A

2016-07-25

We report on generation of ultrabroadband, more than 4 octave spanning supercontinuum in thin CaF₂ crystal, as pumped by intense mid-infrared laser pulses with central wavelength of 2.4 μm. The supercontinuum spectrum covers wavelength range from the ultraviolet to the mid-infrared and its short wavelength side is strongly enhanced by cascaded generation of third, fifth and seventh harmonics. Our results capture the transition from Kerr-dominated to plasma-dominated filamentation regime and uncover that in the latter the spectral superbroadening originates from dramatic plasma-induced compression of the driving pulse, which in turn induces broadening of the harmonics spectra due to cross-phase modulation effects. The experimental measurements are backed up by the numerical simulations based on a nonparaxial unidirectional propagation equation for the electric field of the pulse, which accounts for the cubic nonlinearity-induced effects, and which reproduce the experimental data in great detail.

Surface diffusion of Sb on Ge(111) investigated by second harmonic microscopy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schultz, K.A.

Surface diffusion of Sb on Ge(111) has been measured with the newly-developed technique of second harmonic microscopy. In this method, concentration profiles at submonolayer coverage are imaged directly by second harmonic generation with 5 [mu]m spatial resolution. A Boltzmann-Matano analysis of the concentration profiles yields the coverage dependence of the diffusivity D without parameterization. Experiments were performed at roughly 70% of the bulk melting temperature T[sub m]. In the coverage range of 0 < [theta] < 0.6, the activation energy E[sub diff] remains constant at 47.5 [+-] 1.5 kcal/mol. The corresponding pre-exponential factor decreases from 8.7 [times] 10[sup 3[+-]0.4] tomore » 1.6 [times] 10[sup 2[+-]0.4] cm[sup 2]/sec. The results are explained in terms of a new vacancy model for surface diffusion at high-temperatures. The model accounts semiquantitatively for the large values of E[sub diff] and D[sub o], and suggest that these quantities may be manipulated by bulk doping levels and photon illumination of the surface.« less

Fully integrated reflection-mode photoacoustic, two-photon, and second harmonic generation microscopy in vivo

NASA Astrophysics Data System (ADS)

Song, Wei; Xu, Qiang; Zhang, Yang; Zhan, Yang; Zheng, Wei; Song, Liang

2016-08-01

The ability to obtain comprehensive structural and functional information from intact biological tissue in vivo is highly desirable for many important biomedical applications, including cancer and brain studies. Here, we developed a fully integrated multimodal microscopy that can provide photoacoustic (optical absorption), two-photon (fluorescence), and second harmonic generation (SHG) information from tissue in vivo, with intrinsically co-registered images. Moreover, using a delicately designed optical-acoustic coupling configuration, a high-frequency miniature ultrasonic transducer was integrated into a water-immersion optical objective, thus allowing all three imaging modalities to provide a high lateral resolution of ~290 nm with reflection-mode imaging capability, which is essential for studying intricate anatomy, such as that of the brain. Taking advantage of the complementary and comprehensive contrasts of the system, we demonstrated high-resolution imaging of various tissues in living mice, including microvasculature (by photoacoustics), epidermis cells, cortical neurons (by two-photon fluorescence), and extracellular collagen fibers (by SHG). The intrinsic image co-registration of the three modalities conveniently provided improved visualization and understanding of the tissue microarchitecture. The reported results suggest that, by revealing complementary tissue microstructures in vivo, this multimodal microscopy can potentially facilitate a broad range of biomedical studies, such as imaging of the tumor microenvironment and neurovascular coupling.

Collagenous extracellular matrix of cartilage submitted to mechanical forces studied by second harmonic generation microscopy.

PubMed

Werkmeister, Elisabeth; de Isla, Natalia; Netter, Patrick; Stoltz, Jean-François; Dumas, Dominique

2010-01-01

Osteoarthritis is a degenerative pathology leading to degradation of the extracellular matrix (ECM). Similar effects can be visualized when applying mechanical or biochemical constraints on cartilaginous tissue. Here, we characterized modification of the ECM appearing under mechanical compression and/or biochemical action (hypoxia environment, nitric oxide and collagenase action). In recent decades, multiphoton microscopy has proved its interest for observing living, thick and opaque biological tissues. Thus, the main components of the cartilaginous ECM can be observed without fluorescent labeling. In particular, the collagen network emits strong second harmonic generation (SHG) signal which could be collected at half of the excitation wavelength. Combining autofluorescence and SHG signal detection enables to obtain complementary structural information. Here, we proved that multiphoton microscopy represents an appropriate tool for ex vitro cartilage imaging. First, we showed that SHG signal specifically comes from collagen (collagenase digestion). Further, we verified that the use of an appropriate band-pass filter enables to reject the autofluorescence from the ECM. Once this specificity was shown, we followed modification of the cartilage ECM submitted to mechanical or biochemical constraints (compression, enzymatic digestion). By performing textural analysis of SHG images (Haralick's method), we showed the restructuration of the collagen network according to constraints.

Polarization-modulated second harmonic generation ellipsometric microscopy at video rate.

PubMed

DeWalt, Emma L; Sullivan, Shane Z; Schmitt, Paul D; Muir, Ryan D; Simpson, Garth J

2014-08-19

Fast 8 MHz polarization modulation coupled with analytical modeling, fast beam-scanning, and synchronous digitization (SD) have enabled simultaneous nonlinear optical Stokes ellipsometry (NOSE) and polarized laser transmittance imaging with image acquisition rates up to video rate. In contrast to polarimetry, in which the polarization state of the exiting beam is recorded, NOSE enables recovery of the complex-valued Jones tensor of the sample that describes all polarization-dependent observables of the measurement. Every video-rate scan produces a set of 30 images (10 for each detector with three detectors operating in parallel), each of which corresponds to a different polarization-dependent result. Linear fitting of this image set contracts it down to a set of five parameters for each detector in second harmonic generation (SHG) and three parameters for the transmittance of the incident beam. These parameters can in turn be used to recover the Jones tensor elements of the sample. Following validation of the approach using z-cut quartz, NOSE microscopy was performed for microcrystals of both naproxen and glucose isomerase. When weighted by the measurement time, NOSE microscopy was found to provide a substantial (>7 decades) improvement in the signal-to-noise ratio relative to our previous measurements based on the rotation of optical elements and a 3-fold improvement relative to previous single-point NOSE approaches.

Use of the second harmonic generation microscopy to evaluate chondrogenic differentiation of mesenchymal stem cells for cartilage repair

NASA Astrophysics Data System (ADS)

Bordeaux-Rego, P.; Baratti, M. O.; Duarte, A. S. S.; Ribeiro, T. B.; Andreoli-Risso, M. F.; Vidal, B.; Miranda, J. B.; Adur, J.; de Thomaz, A. A.; Pelegati, V. B.; Costa, F. F.; Carvalho, H. F.; Cesar, C. L.; Luzo, A.; Olalla Saad, S. T.

2012-03-01

Articular cartilage injury remains one of the major concerns in orthopedic surgery. Mesenchymal stem cell (MSC) transplantation has been introduced to avoid some of the side effects and complications of current techniques.. With the aim to evaluate chondrogenic differentiation of mesenchymal stem cells, we used Second Harmonic Generation (SHG) microscopy to analyze the aggregation and orientation of collagen fibrils in the hyaline cartilage of rabbit knees. The experiment was performed using implants with type II collagen hydrogel (a biomaterial that mimics the microenvironment of the cartilage), one implant containing MSC and one other without MSC (control). After 10 weeks, the rabbit knees were dissected and fibril collagen distribution and spatial organization in the extracellular matrix of the lesions were verified by SHG. The result showed significant differences, whereas in histological sections of the cartilaginous lesions with MSC the collagen fibers are organized and regular; in the control sections the collagen fibers are more irregular, with absence of cells. A macroscopic analysis of the lesions confirmed this difference, showing a greater percentage of lesions filling in knees treated with MSC than in the knees used as controls. This study demonstrates that SHG microscopy will be an excellent tool to help in the evaluation of the effectiveness of MSC-based cell therapy for cartilage repair.

Comparison of Cornea Module and DermaInspect for noninvasive imaging of ocular surface pathologies

NASA Astrophysics Data System (ADS)

Steven, Philipp; Müller, Maya; Koop, Norbert; Rose, Christian; Hüttmann, Gereon

2009-11-01

Minimally invasive imaging of ocular surface pathologies aims at securing clinical diagnosis without actual tissue probing. For this matter, confocal microscopy (Cornea Module) is in daily use in ophthalmic practice. Multiphoton microscopy is a new optical technique that enables high-resolution imaging and functional analysis of living tissues based on tissue autofluorescence. This study was set up to compare the potential of a multiphoton microscope (DermaInspect) to the Cornea Module. Ocular surface pathologies such as pterygia, papillomae, and nevi were investigated in vivo using the Cornea Module and imaged immediately after excision by DermaInspect. Two excitation wavelengths, fluorescence lifetime imaging and second-harmonic generation (SHG), were used to discriminate different tissue structures. Images were compared with the histopathological assessment of the samples. At wavelengths of 730 nm, multiphoton microscopy exclusively revealed cellular structures. Collagen fibrils were specifically demonstrated by second-harmonic generation. Measurements of fluorescent lifetimes enabled the highly specific detection of goblet cells, erythrocytes, and nevus-cell clusters. At the settings used, DermaInspect reaches higher resolutions than the Cornea Module and obtains additional structural information. The parallel detection of multiphoton excited autofluorescence and confocal imaging could expand the possibilities of minimally invasive investigation of the ocular surface toward functional analysis at higher resolutions.

Automated biphasic morphological assessment of hepatitis B-related liver fibrosis using second harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Wang, Tong-Hong; Chen, Tse-Ching; Teng, Xiao; Liang, Kung-Hao; Yeh, Chau-Ting

2015-08-01

Liver fibrosis assessment by biopsy and conventional staining scores is based on histopathological criteria. Variations in sample preparation and the use of semi-quantitative histopathological methods commonly result in discrepancies between medical centers. Thus, minor changes in liver fibrosis might be overlooked in multi-center clinical trials, leading to statistically non-significant data. Here, we developed a computer-assisted, fully automated, staining-free method for hepatitis B-related liver fibrosis assessment. In total, 175 liver biopsies were divided into training (n = 105) and verification (n = 70) cohorts. Collagen was observed using second harmonic generation (SHG) microscopy without prior staining, and hepatocyte morphology was recorded using two-photon excitation fluorescence (TPEF) microscopy. The training cohort was utilized to establish a quantification algorithm. Eleven of 19 computer-recognizable SHG/TPEF microscopic morphological features were significantly correlated with the ISHAK fibrosis stages (P < 0.001). A biphasic scoring method was applied, combining support vector machine and multivariate generalized linear models to assess the early and late stages of fibrosis, respectively, based on these parameters. The verification cohort was used to verify the scoring method, and the area under the receiver operating characteristic curve was >0.82 for liver cirrhosis detection. Since no subjective gradings are needed, interobserver discrepancies could be avoided using this fully automated method.

Second harmonic generation microscopy analysis of extracellular matrix changes in human idiopathic pulmonary fibrosis

PubMed Central

Tilbury, Karissa; Hocker, James; Wen, Bruce L.; Sandbo, Nathan; Singh, Vikas; Campagnola, Paul J.

2014-01-01

Abstract. Patients with idiopathic fibrosis (IPF) have poor long-term survival as there are limited diagnostic/prognostic tools or successful therapies. Remodeling of the extracellular matrix (ECM) has been implicated in IPF progression; however, the structural consequences on the collagen architecture have not received considerable attention. Here, we demonstrate that second harmonic generation (SHG) and multiphoton fluorescence microscopy can quantitatively differentiate normal and IPF human tissues. For SHG analysis, we developed a classifier based on wavelet transforms, principle component analysis, and a K-nearest-neighbor algorithm to classify the specific alterations of the collagen structure observed in IPF tissues. The resulting ROC curves obtained by varying the numbers of principal components and nearest neighbors yielded accuracies of >95%. In contrast, simpler metrics based on SHG intensity and collagen coverage in the image provided little or no discrimination. We also characterized the change in the elastin/collagen balance by simultaneously measuring the elastin autofluorescence and SHG intensities and found that the IPF tissues were less elastic relative to collagen. This is consistent with known mechanical consequences of the disease. Understanding ECM remodeling in IPF via nonlinear optical microscopy may enhance our ability to differentiate patients with rapid and slow progression and, thus, provide better prognostic information. PMID:25134793

Automated biphasic morphological assessment of hepatitis B-related liver fibrosis using second harmonic generation microscopy.

PubMed

Wang, Tong-Hong; Chen, Tse-Ching; Teng, Xiao; Liang, Kung-Hao; Yeh, Chau-Ting

2015-08-11

Liver fibrosis assessment by biopsy and conventional staining scores is based on histopathological criteria. Variations in sample preparation and the use of semi-quantitative histopathological methods commonly result in discrepancies between medical centers. Thus, minor changes in liver fibrosis might be overlooked in multi-center clinical trials, leading to statistically non-significant data. Here, we developed a computer-assisted, fully automated, staining-free method for hepatitis B-related liver fibrosis assessment. In total, 175 liver biopsies were divided into training (n = 105) and verification (n = 70) cohorts. Collagen was observed using second harmonic generation (SHG) microscopy without prior staining, and hepatocyte morphology was recorded using two-photon excitation fluorescence (TPEF) microscopy. The training cohort was utilized to establish a quantification algorithm. Eleven of 19 computer-recognizable SHG/TPEF microscopic morphological features were significantly correlated with the ISHAK fibrosis stages (P < 0.001). A biphasic scoring method was applied, combining support vector machine and multivariate generalized linear models to assess the early and late stages of fibrosis, respectively, based on these parameters. The verification cohort was used to verify the scoring method, and the area under the receiver operating characteristic curve was >0.82 for liver cirrhosis detection. Since no subjective gradings are needed, interobserver discrepancies could be avoided using this fully automated method.

Compact single-pass X-ray FEL with harmonic multiplication cascades

NASA Astrophysics Data System (ADS)

Zhukovsky, K.

2018-07-01

The generation of X-ray radiation in cascaded single-pass free electron laser (FEL), which amplifies high harmonics of a two-frequency undulator, is studied. Power dynamics of FEL harmonics is explored with the help of the phenomenological model of a single pass FEL. The model describes both linear and non-linear harmonic generation, starting from a coherent seed laser and initial shot noise with account for main loss factors for each harmonic in each cascade individually: the energy spread and beam divergence, the coupling losses between FEL cascades, the diffraction etc. The model was validated with the experiment and with relevant 3-D simulations. It is employed for modeling the cascaded FELs with harmonic multiplication and analyzing the evolution of FEL harmonic power with the aim to obtain the maximum high harmonic power in the X-ray band at the shortest possible FEL length with the lowest possible seed frequency. The advantages of two-frequency undulators in HGHG FELs are elucidated. The requirements for the electron beam are studied; the need for low energy spread is evidenced: our evaluations yield σe < 2 × 10-4. Several cascaded HGHG FELs with two-frequency undulators are modeled. Generation of soft X-ray radiation at λ = 2 . 71 nm, reaching ∼50 MW power with I0 ∼ 100 A in a cascaded FEL at just 40 m with 13.51 nm seed, matching peak reflectivity of Mo/Si, is demonstrated. The generation of 40 MW radiation power at λ = 2 . 27 nm with the beam current I0 ∼ 100 A, energy E = 950 MeV and the energy spread σe = 2 × 10-4 is studied, using second and third harmonics in three-stage 45 m long FEL. The multistage FEL is modeled for generating radiation in nanometer band: ∼40 MW power at λ ∼ 2 . 6 nm with I0 ∼ 175 A current in just ∼40 m long FEL with commercially available F2 excimer UV laser seed at 157 nm. The peak radiation power rises to ∼0.5 GW for ∼1 kA beam current.

Ultrafast optical pulse delivery with fibers for nonlinear microscopy

PubMed Central

Kim, Daekeun; Choi, Heejin; Yazdanfar, Siavash; So, Peter T. C.

2008-01-01

Nonlinear microscopies including multiphoton excitation fluorescence microscopy and multiple-harmonic generation microscopy have recently gained popularity for cellular and tissue imaging. The optimization of these imaging methods for minimally invasive use will require optical fibers to conduct light into tight space where free space delivery is difficult. The delivery of high peak power laser pulses with optical fibers is limited by dispersion resulting from nonlinear refractive index responses. In this paper, we characterize a variety of commonly used optical fibers in terms of how they affect pulse profile and imaging performance of nonlinear microscopy; the following parameters are quantified: spectral bandwidth and temporal pulse width, two-photon excitation efficiency, and optical resolution. A theoretical explanation for the measured performance of these is also provided. PMID:18816597

On mapping subangstrom electron clouds with force microscopy.

PubMed

Wright, C Alan; Solares, Santiago D

2011-11-09

In 2004 Hembacher et al. (Science 2004, 305, 380-383) reported simultaneous higher-harmonics atomic force mocroscopy (AFM)/scanning tunneling microscopy (STM) images acquired while scanning a graphite surface with a tungsten tip. They interpreted the observed subatomic features in the AFM images as the signature of lobes of increased electron density at the tungsten tip apex. Although these intriguing images have stirred controversy, an in-depth theoretical feasibility study has not yet been produced. Here we report on the development of a method for simulating higher harmonics AFM images and its application to the same system. Our calculations suggest that four lobes of increased electron density are expected to be present at a W(001) tip apex atom and that the corresponding higher harmonics AFM images of graphite can exhibit 4-fold symmetry features. Despite these promising results, open questions remain since the calculated amplitudes of the higher harmonics generated by the short-range forces are on the order of hundredths of picometers, leading to very small corrugations in the theoretical images. Additionally, the complex, intermittent nature of the tip-sample interaction, which causes constant readjustment of the tip and sample orbitals as the tip approaches and retracts from the surface, prevents a direct quantitative connection between the electron density and the AFM image features.

Linear transformer driver for pulse generation with fifth harmonic

DOEpatents

Mazarakis, Michael G.; Kim, Alexander A.; Sinebryukhov, Vadim A.; Volkov, Sergey N.; Kondratiev, Sergey S.; Alexeenko, Vitaly M.; Bayol, Frederic; Demol, Gauthier; Stygar, William A.; Leckbee, Joshua; Oliver, Bryan V.; Kiefer, Mark L.

2017-03-21

A linear transformer driver includes at least one ferrite ring positioned to accept a load. The linear transformer driver also includes a first, second, and third power delivery module. The first power delivery module sends a first energy in the form of a first pulse to the load. The second power delivery module sends a second energy in the form of a second pulse to the load. The third power delivery module sends a third energy in the form of a third pulse to the load. The linear transformer driver is configured to form a flat-top pulse by the superposition of the first, second, and third pulses. The first, second, and third pulses have different frequencies.

Multimodal fiber source for nonlinear microscopy based on a dissipative soliton laser

PubMed Central

Lamb, Erin S.; Wise, Frank W.

2015-01-01

Recent developments in high energy femtosecond fiber lasers have enabled robust and lower-cost sources for multiphoton-fluorescence and harmonic-generation imaging. However, picosecond pulses are better suited for Raman scattering microscopy, so the ideal multimodal source for nonlinear microcopy needs to provide both durations. Here we present spectral compression of a high-power femtosecond fiber laser as a route to producing transform-limited picosecond pulses. These pulses pump a fiber optical parametric oscillator to yield a robust fiber source capable of providing the synchronized picosecond pulse trains needed for Raman scattering microscopy. Thus, this system can be used as a multimodal platform for nonlinear microscopy techniques. PMID:26417497

InSe monolayer: synthesis, structure and ultra-high second-harmonic generation

NASA Astrophysics Data System (ADS)

Zhou, Jiadong; Shi, Jia; Zeng, Qingsheng; Chen, Yu; Niu, Lin; Liu, Fucai; Yu, Ting; Suenaga, Kazu; Liu, Xinfeng; Lin, Junhao; Liu, Zheng

2018-04-01

III–IV layered materials such as indium selenide have excellent photoelectronic properties. However, synthesis of materials in such group, especially with a controlled thickness down to monolayer, still remains challenging. Herein, we demonstrate the successful synthesis of monolayer InSe by physical vapor deposition (PVD) method. The high quality of the sample was confirmed by complementary characterization techniques such as Raman spectroscopy, atomic force microscopy (AFM) and high resolution annular dark field scanning transmission electron microscopy (ADF-STEM). We found the co-existence of different stacking sequence (β- and γ-InSe) in the same flake with a sharp grain boundary in few-layered InSe. Edge reconstruction is also observed in monolayer InSe, which has a distinct atomic structure from the bulk lattice. Moreover, we discovered that the second-harmonic generation (SHG) signal from monolayer InSe shows large optical second-order susceptibility that is 1–2 orders of magnitude higher than MoS2, and even 3 times of the largest value reported in monolayer GaSe. These results make atom-thin InSe a promising candidate for optoelectronic and photosensitive device applications.

Second harmonic generation microscopy of the living human cornea

NASA Astrophysics Data System (ADS)

Artal, Pablo; Ávila, Francisco; Bueno, Juan

2018-02-01

Second Harmonic Generation (SHG) microscopy provides high-resolution structural imaging of the corneal stroma without the need of labelling techniques. This powerful tool has never been applied to living human eyes so far. Here, we present a new compact SHG microscope specifically developed to image the structural organization of the corneal lamellae in living healthy human volunteers. The research prototype incorporates a long-working distance dry objective that allows non-contact three-dimensional SHG imaging of the cornea. Safety assessment and effectiveness of the system were firstly tested in ex-vivo fresh eyes. The maximum average power of the used illumination laser was 20 mW, more than 10 times below the maximum permissible exposure (according to ANSI Z136.1-2000). The instrument was successfully employed to obtain non-contact and non-invasive SHG of the living human eye within well-established light safety limits. This represents the first recording of in vivo SHG images of the human cornea using a compact multiphoton microscope. This might become an important tool in Ophthalmology for early diagnosis and tracking ocular pathologies.

Enhancing resolution and contrast in second-harmonic generation microscopy using an advanced maximum likelihood estimation restoration method

NASA Astrophysics Data System (ADS)

Sivaguru, Mayandi; Kabir, Mohammad M.; Gartia, Manas Ranjan; Biggs, David S. C.; Sivaguru, Barghav S.; Sivaguru, Vignesh A.; Berent, Zachary T.; Wagoner Johnson, Amy J.; Fried, Glenn A.; Liu, Gang Logan; Sadayappan, Sakthivel; Toussaint, Kimani C.

2017-02-01

Second-harmonic generation (SHG) microscopy is a label-free imaging technique to study collagenous materials in extracellular matrix environment with high resolution and contrast. However, like many other microscopy techniques, the actual spatial resolution achievable by SHG microscopy is reduced by out-of-focus blur and optical aberrations that degrade particularly the amplitude of the detectable higher spatial frequencies. Being a two-photon scattering process, it is challenging to define a point spread function (PSF) for the SHG imaging modality. As a result, in comparison with other two-photon imaging systems like two-photon fluorescence, it is difficult to apply any PSF-engineering techniques to enhance the experimental spatial resolution closer to the diffraction limit. Here, we present a method to improve the spatial resolution in SHG microscopy using an advanced maximum likelihood estimation (AdvMLE) algorithm to recover the otherwise degraded higher spatial frequencies in an SHG image. Through adaptation and iteration, the AdvMLE algorithm calculates an improved PSF for an SHG image and enhances the spatial resolution by decreasing the full-width-at-halfmaximum (FWHM) by 20%. Similar results are consistently observed for biological tissues with varying SHG sources, such as gold nanoparticles and collagen in porcine feet tendons. By obtaining an experimental transverse spatial resolution of 400 nm, we show that the AdvMLE algorithm brings the practical spatial resolution closer to the theoretical diffraction limit. Our approach is suitable for adaptation in micro-nano CT and MRI imaging, which has the potential to impact diagnosis and treatment of human diseases.

Second harmonic generation quantitative measurements on collagen fibrils through correlation to electron microscopy

NASA Astrophysics Data System (ADS)

Bancelin, S.; Aimé, C.; Gusachenko, I.; Kowalczuk, L.; Latour, G.; Coradin, T.; Schanne-Klein, M.-C.

2015-03-01

Type I collagen is a major structural protein in mammals that shows highly structured macromolecular organizations specific to each tissue. This biopolymer is synthesized as triple helices, which self-assemble into fibrils (Ø =10-300 nm) and further form various 3D organization. In recent years, Second Harmonic Generation (SHG) microscopy has emerged as a powerful technique to probe in situ the fibrillar collagenous network within tissues. However, this optical technique cannot resolve most of the fibrils and is a coherent process, which has impeded quantitative measurements of the fibril diameter so far. In this study, we correlated SHG microscopy with Transmission Electron Microscopy to determine the sensitivity of SHG microscopy and to calibrate SHG signals as a function of the fibril diameter in reconstructed collagen gels. To that end, we synthetized isolated fibrils with various diameters and successfully imaged the very same fibrils with both techniques, down to 30 nm diameter. We observed that SHG signals scaled as the fourth power of the fibril diameter, as expected from analytical and numerical calculations. This calibration was then applied to diabetic rat cornea in which we successfully recovered the diameter of hyperglycemia-induced fibrils in the Descemet's membrane without having to resolve them. Finally we derived the first hyperpolarizability from a single collagen triple helix which validates the bottom-up approach used to calculate the non-linear response at the fibrillar scale and denotes a parallel alignment of triple helices within the fibrils. These results represent a major step towards quantitative SHG imaging of nm-sized collagen fibrils.

Low complexity feature extraction for classification of harmonic signals

NASA Astrophysics Data System (ADS)

William, Peter E.

In this dissertation, feature extraction algorithms have been developed for extraction of characteristic features from harmonic signals. The common theme for all developed algorithms is the simplicity in generating a significant set of features directly from the time domain harmonic signal. The features are a time domain representation of the composite, yet sparse, harmonic signature in the spectral domain. The algorithms are adequate for low-power unattended sensors which perform sensing, feature extraction, and classification in a standalone scenario. The first algorithm generates the characteristic features using only the duration between successive zero-crossing intervals. The second algorithm estimates the harmonics' amplitudes of the harmonic structure employing a simplified least squares method without the need to estimate the true harmonic parameters of the source signal. The third algorithm, resulting from a collaborative effort with Daniel White at the DSP Lab, University of Nebraska-Lincoln, presents an analog front end approach that utilizes a multichannel analog projection and integration to extract the sparse spectral features from the analog time domain signal. Classification is performed using a multilayer feedforward neural network. Evaluation of the proposed feature extraction algorithms for classification through the processing of several acoustic and vibration data sets (including military vehicles and rotating electric machines) with comparison to spectral features shows that, for harmonic signals, time domain features are simpler to extract and provide equivalent or improved reliability over the spectral features in both the detection probabilities and false alarm rate.

Dual-pulse frequency compounded superharmonic imaging.

PubMed

van Neer, Paul L M J; Danilouchkine, Mikhail G; Matte, Guillaume M; van der Steen, Anton F W; de Jong, Nico

2011-11-01

Tissue second-harmonic imaging is currently the default mode in commercial diagnostic ultrasound systems. A new modality, superharmonic imaging (SHI), combines the third through fifth harmonics originating from nonlinear wave propagation through tissue. SHI could further improve the resolution and quality of echographic images. The superharmonics have gaps between the harmonics because the transducer has a limited bandwidth of about 70% to 80%. This causes ghost reflection artifacts in the superharmonic echo image. In this work, a new dual-pulse frequency compounding (DPFC) method to eliminate these artifacts is introduced. In the DPFC SHI method, each trace is constructed by summing two firings with slightly different center frequencies. The feasibility of the method was established using a single-element transducer. Its acoustic field was modeled in KZK simulations and compared with the corresponding measurements obtained with a hydrophone apparatus. Subsequently, the method was implemented on and optimized for a setup consisting of an interleaved phased-array transducer (44 elements at 1 MHz and 44 elements at 3.7 MHz, optimized for echocardiography) and a programmable ultrasound system. DPFC SHI effectively suppresses the ghost reflection artifacts associated with imaging using multiple harmonics. Moreover, compared with the single-pulse third harmonic, DPFC SHI improved the axial resolution by 3.1 and 1.6 times at the -6-dB and -20-dB levels, respectively. Hence, DPFC offers the possibility of generating harmonic images of a higher quality at a cost of a moderate frame rate reduction.

Nonlinear symmetry breaking in photometamaterials

NASA Astrophysics Data System (ADS)

Gorlach, Maxim A.; Dobrykh, Dmitry A.; Slobozhanyuk, Alexey P.; Belov, Pavel A.; Lapine, Mikhail

2018-03-01

We design and analyze theoretically photometamaterials with each meta-atom containing both photodiode and light-emitting diode. Illumination of the photodiode by the light-emitting diode gives rise to an additional optical feedback within each unit cell, which strongly affects resonant properties and nonlinear response of the meta-atom. In particular, we demonstrate that inversion symmetry breaking occurs upon a certain threshold magnitude of the incident wave intensity resulting in an abrupt emergence of second-harmonic generation, which was not originally available, as well as in the reduced third-harmonic signal.

Phase noise mitigation of QPSK signal utilizing phase-locked multiplexing of signal harmonics and amplitude saturation.

PubMed

Mohajerin-Ariaei, Amirhossein; Ziyadi, Morteza; Chitgarha, Mohammad Reza; Almaiman, Ahmed; Cao, Yinwen; Shamee, Bishara; Yang, Jeng-Yuan; Akasaka, Youichi; Sekiya, Motoyoshi; Takasaka, Shigehiro; Sugizaki, Ryuichi; Touch, Joseph D; Tur, Moshe; Langrock, Carsten; Fejer, Martin M; Willner, Alan E

2015-07-15

We demonstrate an all-optical phase noise mitigation scheme based on the generation, delay, and coherent summation of higher order signal harmonics. The signal, its third-order harmonic, and their corresponding delayed variant conjugates create a staircase phase-transfer function that quantizes the phase of quadrature-phase-shift-keying (QPSK) signal to mitigate phase noise. The signal and the harmonics are automatically phase-locked multiplexed, avoiding the need for phase-based feedback loop and injection locking to maintain coherency. The residual phase noise converts to amplitude noise in the quantizer stage, which is suppressed by parametric amplification in the saturation regime. Phase noise reduction of ∼40% and OSNR-gain of ∼3  dB at BER 10(-3) are experimentally demonstrated for 20- and 30-Gbaud QPSK input signals.

Unlocking higher harmonics in atomic force microscopy with gentle interactions.

PubMed

Santos, Sergio; Barcons, Victor; Font, Josep; Verdaguer, Albert

2014-01-01

In dynamic atomic force microscopy, nanoscale properties are encoded in the higher harmonics. Nevertheless, when gentle interactions and minimal invasiveness are required, these harmonics are typically undetectable. Here, we propose to externally drive an arbitrary number of exact higher harmonics above the noise level. In this way, multiple contrast channels that are sensitive to compositional variations are made accessible. Numerical integration of the equation of motion shows that the external introduction of exact harmonic frequencies does not compromise the fundamental frequency. Thermal fluctuations are also considered within the detection bandwidth of interest and discussed in terms of higher-harmonic phase contrast in the presence and absence of an external excitation of higher harmonics. Higher harmonic phase shifts further provide the means to directly decouple the true topography from that induced by compositional heterogeneity.

Structural Investigation of Biological and Semiconductor Nanostructures with Nonlinear Multicontrast Microscopy

NASA Astrophysics Data System (ADS)

Cisek, Richard

Physical and functional properties of advanced nano-composite materials and biological structures are determined by self-organized atoms and molecules into nanostructures and in turn by microscopic organization of the nanostructures into assemblies of higher structural complexity. Therefore, microscopes are indispensable tools for structural investigations at various levels of organization. In this work, novel nonlinear optical microscopy methods were developed to non-invasively study structural organization at the nanoscopic and microscopic levels. Atomic organization of semiconductor nanowires, molecular organization of amylose biocrystallites in starch granules, and microscopic organization of several photosynthetic organisms was elucidated. The structure of ZnSe nanowires, key components in many modern nanodevices, was investigated using polarization harmonic generation microscopy. Based on nonlinear optical properties of the different crystal lattices, zinc blende and wurtzite nanowires were differentiated, and the three-dimensional orientation of the zinc blende nanowires could be found. The structure of starch granules, a model biocrystal, important in food as well as health sciences, was also investigated using polarization harmonic microscopy. The study was combined with ab initio calculations using the crystal structures of amylose A and B, revealing that second harmonic signals originate from the hydroxide and hydrogen bonds in the starch granules. Visualization of several photosynthetic organisms including the green algae, Chlamydomonas reinhardtii, two species of cyanobacteria, Leptolyngbya sp. and Anabaena sp., aggregates of light-harvesting pigment-protein complexes as well as chloroplasts from green plants were also explored, revealing that future nonlinear microscopy applications could include structural studies of cell walls, the Chlamydomonas eyespot, and photosynthetic membranes. In this study, several nonlinear optical microscopy modalities were developed for quantitative structural investigations of nano and micro-sized architectures. Non-invasive extraction of crystallographic information in microscopic samples will have a number of potential benefits, for example, in clinical applications, allowing observations of disease states inside tissues without the need for biopsy. Industrial nanotechnology will benefit from fast determination of nanostructures with nonlinear microscopy that will improve quality of nanodevices.

High efficiency and output power from second- and third-harmonic millimeter-wave InP-TED oscillators at frequencies above 170 GHz

NASA Astrophysics Data System (ADS)

Rydberg, Anders

1990-10-01

InP TED (transferred electron device) oscillators have been experimentally investigated for frequencies between 170 and 279 GHz. It has been found that output powers of more than 7 and 0.2 mW are possible at 180 and 272 GHz using second- and third-harmonic mode operation, respectively. Conversion efficiencies of more than 13 percent and 0.3 percent between fundamental and second harmonic and fundamental and third harmonic, respectively, have been found. The conversion efficiencies are comparable to GaAs TEDs. The output powers, conversion efficiencies, and tuning ranges (more than 22 percent) are the largest reported for InP TEDs at these frequencies. The output power at third harmonic was sufficient for supplying a superconducting mixer with local oscillator power.

Generation of multicolor vacuum ultraviolet pulses through four-wave sum-frequency mixing in argon

NASA Astrophysics Data System (ADS)

Shi, Liping; Li, Wenxue; Zhou, Hui; Wang, Di; Ding, Liang'en; Zeng, Heping

2013-11-01

We demonstrate efficient generation of multicolor vacuum ultraviolet pulses with excellent mode quality through χ(3)-based four-wave sum-frequency mixing and third-order harmonic generation of 400- and 267-nm femtosecond laser pulses in argon gas. The χ(3)-based nonlinear optical processes were optimized with appropriate control of gas pressure and group velocity delay between the driving pulses. Furthermore, the pulse breakup effects were observed for tightly focused ultraviolet pulses.

Hyperspectral imaging with laser-scanning sum-frequency generation microscopy

PubMed Central

Hanninen, Adam; Shu, Ming Wai; Potma, Eric O.

2017-01-01

Vibrationally sensitive sum-frequency generation (SFG) microscopy is a chemically selective imaging technique sensitive to non-centrosymmetric molecular arrangements in biological samples. The routine use of SFG microscopy has been hampered by the difficulty of integrating the required mid-infrared excitation light into a conventional, laser-scanning nonlinear optical (NLO) microscope. In this work, we describe minor modifications to a regular laser-scanning microscope to accommodate SFG microscopy as an imaging modality. We achieve vibrationally sensitive SFG imaging of biological samples with sub-μm resolution at image acquisition rates of 1 frame/s, almost two orders of magnitude faster than attained with previous point-scanning SFG microscopes. Using the fast scanning capability, we demonstrate hyperspectral SFG imaging in the CH-stretching vibrational range and point out its use in the study of molecular orientation and arrangement in biologically relevant samples. We also show multimodal imaging by combining SFG microscopy with second-harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) on the same imaging platfrom. This development underlines that SFG microscopy is a unique modality with a spatial resolution and image acquisition time comparable to that of other NLO imaging techniques, making point-scanning SFG microscopy a valuable member of the NLO imaging family. PMID:28966861

Development of high repetition rate nitric oxide planar laser induced fluorescence imaging

NASA Astrophysics Data System (ADS)

Jiang, Naibo

This thesis has documented the development of a MHz repitition rate pulse burst laser system. Second harmonic and third harmonic efficiencies are improved by adding a Phase Conjugate Mirror to the system. Some high energy fundamental, second harmonic, and third harmonic burst sequences consisting of 1--12 pulses separated in time by between 4 and 12 microseconds are now routinely obtained. The reported burst envelopes are quite uniform. We have also demonstrated the ability to generate ultra-high frequency sequences of broadly wavelength tunable, high intensity laser pulses using a home built injection seeded Optical Parametric Oscillator (OPO), pumped by the second and third harmonic output of the pulse burst laser. Typical OPO output burst sequences consist of 6--10 pulses, separated in time by between 6 and 10 microseconds. With third harmonic pumping of the OPO system, we studied four conditions, two-crystal Singly Resonant OPO (SRO) cavity, three-crystal OPO cavity, single pass two-crystal Doubly Resonant OPO (DRO) cavity and double pass two-crystal OPO cavity. The double pass two-crystal OPO cavity gives the best operation in burst mode. For single pass OPO, the average total OPO conversion efficiency is approximately 25%. For double pass OPO, the average total OPO conversion efficiency is approximately 35%. As a preliminary work, we studied 532nm pumping of a single crystal OPO cavity. With single pulse pumping, the conversion efficiency can reach 30%. For both 355nm and 532nm pumping OPO, we have demonstrated injection seeding. The OPO output light linewidth is significantly narrowed. Some preliminary etalon traces are also reported. By mixing the OPO signal output at 622nm with residual third harmonic at 355nm, we obtained 226nm burst sequences with average pulse energy of ˜0.2 mJ. Injection seeding of the OPO increases the energy achieved by a factor of ˜2. 226nm burst sequences with reasonably uniform burst envelopes are reported. Using the system we have obtained, for the first time by any known optical method, Planar Laser Induced Fluorescence (PLIF) image sequences at ultrahigh (≥100kHz) frame rates, in particular NO PLIF image sequences, have been obtained in a Mach 2 jet. We also studied the possibility of utilizing a 250 kHz pulsed Nd:YVO 4 laser as the master oscillator. 10-pulse-10-mus spacing burst sequences with reasonably uniform burst envelope have been obtained. The total energy of the burst sequence is ˜2.5J.

Nanotwin Detection and Domain Polarity Determination via Optical Second Harmonic Generation Polarimetry.

PubMed

Ren, Ming-Liang; Agarwal, Rahul; Nukala, Pavan; Liu, Wenjing; Agarwal, Ritesh

2016-07-13

We demonstrate that optical second harmonic generation (SHG) can be utilized to determine the exact nature of nanotwins in noncentrosymmetric crystals, which is challenging to resolve via conventional transmission electron or scanned probe microscopies. Using single-crystalline nanotwinned CdTe nanobelts and nanowires as a model system, we show that SHG polarimetry can distinguish between upright (Cd-Te bonds) and inverted (Cd-Cd or Te-Te bonds) twin boundaries in the system. Inverted twin boundaries are generally not reported in nanowires due to the lack of techniques and complexity associated with the study of the nature of such defects. Precise characterization of the nature of defects in nanocrystals is required for deeper understanding of their growth and physical properties to enable their application in future devices.

Laterally azo-bridged h-shaped ferroelectric dimesogens for second-order nonlinear optics: ferroelectricity and second harmonic generation.

PubMed

Zhang, Yongqiang; Martinez-Perdiguero, Josu; Baumeister, Ute; Walker, Christopher; Etxebarria, Jesus; Prehm, Marko; Ortega, Josu; Tschierske, Carsten; O'Callaghan, Michael J; Harant, Adam; Handschy, Mark

2009-12-30

Two classes of laterally azo-bridged H-shaped ferroelectric liquid crystals (FLCs), incorporating azobenzene and disperse red 1 (DR-1) chromophores along the FLC polar axes, were synthesized and characterized by polarized light microscopy, differential scanning calorimetry, 2D X-ray diffraction analysis, and electro-optical investigations. They represent the first H-shaped FLC materials exhibiting the ground-state, thermodynamically stable enantiotropic SmC* phase, i.e., ground-state ferroelectricity. Second harmonic generation measurements of one compound incorporating a DR-1 chromophore at the incident wavelength of 1064 nm give a nonlinear coefficient of d(22) = 17 pm/V, the largest nonlinear optics coefficient reported to date for calamitic FLCs. This value enables viable applications of FLCs in nonlinear optics.

Exploiting elastic anharmonicity in aluminum nitride matrix for phase-synchronous frequency reference generation

NASA Astrophysics Data System (ADS)

Ghatge, Mayur; Tabrizian, Roozbeh

2018-03-01

A matrix of aluminum-nitride (AlN) waveguides is acoustically engineered to realize electrically isolated phase-synchronous frequency references through nonlinear wave-mixing. AlN rectangular waveguides are cross-coupled through a periodically perforated plate that is engineered to have a wide acoustic bandgap around a desirable frequency ( f1≈509 MHz). While the coupling plate isolates the matrix from resonant vibrations of individual waveguide constituents at f1, it is transparent to the third-order harmonic waves (3f1) that are generated through nonlinear wave-mixing. Therefore, large-signal excitation of the f1 mode in a constituent waveguide generates acoustic waves at 3f1 with an efficiency defined by elastic anharmonicity of the AlN film. The phase-synchronous propagation of the third harmonic through the matrix is amplified by a high quality-factor resonance mode at f2≈1529 MHz, which is sufficiently close to 3f1 (f2 ≅ 3f1). Such an architecture enables realization of frequency-multiplied and phase-synchronous, yet electrically and spectrally isolated, references for multi-band/carrier and spread-spectrum wireless communication systems.

Second harmonic detection in the electrochemical strain microscopy of Ag-ion conducting glass

DOE PAGES

Yang, Sangmo; Okatan, Mahmut Baris; Paranthaman, Mariappan Parans; ...

2014-11-14

The first and second harmonic electromechanical responses and their cross-correlation in Ag-ion conducting glass were investigated using band-excitation electrochemical strain microscopy (ESM). Consecutive ESM images with increasing magnitudes of the applied AC voltage allowed observation of not only reversible surface displacement but also irreversible silver nanoparticle formation above a certain threshold voltage. The second harmonic ESM response was anticorrelated with the first harmonic response in many local regions. Furthermore, the nucleation sites of silver nanoparticles were closely related to the anti-correlated regions, specifically, with low second harmonic and high first harmonic ESM responses. The possible origins of the second harmonicmore » ESM response are discussed.« less

Hybrid label-free multiphoton and optoacoustic microscopy (MPOM)

NASA Astrophysics Data System (ADS)

Soliman, Dominik; Tserevelakis, George J.; Omar, Murad; Ntziachristos, Vasilis

2015-07-01

Many biological applications require a simultaneous observation of different anatomical features. However, unless potentially harmful staining of the specimens is employed, individual microscopy techniques do generally not provide multi-contrast capabilities. We present a hybrid microscope integrating optoacoustic microscopy and multiphoton microscopy, including second-harmonic generation, into a single device. This combined multiphoton and optoacoustic microscope (MPOM) offers visualization of a broad range of structures by employing different contrast mechanisms and at the same time enables pure label-free imaging of biological systems. We investigate the relative performance of the two microscopy modalities and demonstrate their multi-contrast abilities through the label-free imaging of a zebrafish larva ex vivo, simultaneously visualizing muscles and pigments. This hybrid microscopy application bears great potential for developmental biology studies, enabling more comprehensive information to be obtained from biological specimens without the necessity of staining.

Polarization-resolved SHG microscopy in cardiac hypertrophy study (Conference Presentation)

NASA Astrophysics Data System (ADS)

Wang, Zhonghai; Yuan, Cai; Shao, Yonghong; Bradshaw, Amy D.; Borg, Thomas K.; Gao, Bruce Z.

2017-02-01

Cardiac hypertrophy, a process initiated by mechanical alterations, is hypothesized to cause long-term molecular-level alteration in the sarcomere lattice, which is the main force-generating component in the heart muscle. This molecular-level alteration is beyond the resolving capacity of common light microscopy. Second harmonic generation (SHG) microscopy has unique capability for visualizing ordered molecular structures in biological tissues without labeling. Combined with polarization imaging technique, SHG microscopy is able to extract structural details of myosin at the molecular level so as to reveal molecular-level alterations that occur during hypertrophy. The myosin filaments are believed to possess C6 symmetry; thus, the nonlinear polarization response relationship between generated second harmonic light I^2ωand incident fundamental light I^ω is determined by nonlinear coefficients, χ_15, χ_31 and χ_33. χ_31/χ_15 is believed to be an indicator of the molecular symmetry of myosin filament, whileχ_33/χ_15represents the intramyosin orientation angle of the double helix. By changing the polarization of the incident light and evaluating the corresponding SHG signals, the molecular structure of the myosin, reflected by the χ coefficients, can be revealed. With this method, we studied the structural properties of heart tissues in different conditions, including those in normal, physiologically hypertrophic (heart tissue from postpartum female rats), and pathologically hypertrophic (heart tissue from transverse-aorta constricted rats) conditions. We found that ratios of χ_31/χ_15 showed no significant difference between heart tissues from different conditions; their values were all close to 1, which demonstrated that Kleinman symmetry held for all conditions. Ratios of χ_33/χ_15 from physiologically or pathologically hypertrophic heart tissues were raised and showed significant difference from those from normal heart tissues, which indicated that the intramyosin orientation angle of the double helix was altered when heart tissues hypertrophied. Polarization-resolved SHG microscopy permitted us to study heart tissues at the molecular level and may serve as a diagnostic tool for cardiac hypertrophy.

Synchronous-digitization for Video Rate Polarization Modulated Beam Scanning Second Harmonic Generation Microscopy.

PubMed

Sullivan, Shane Z; DeWalt, Emma L; Schmitt, Paul D; Muir, Ryan M; Simpson, Garth J

2015-03-09

Fast beam-scanning non-linear optical microscopy, coupled with fast (8 MHz) polarization modulation and analytical modeling have enabled simultaneous nonlinear optical Stokes ellipsometry (NOSE) and linear Stokes ellipsometry imaging at video rate (15 Hz). NOSE enables recovery of the complex-valued Jones tensor that describes the polarization-dependent observables, in contrast to polarimetry, in which the polarization stated of the exciting beam is recorded. Each data acquisition consists of 30 images (10 for each detector, with three detectors operating in parallel), each of which corresponds to polarization-dependent results. Processing of this image set by linear fitting contracts down each set of 10 images to a set of 5 parameters for each detector in second harmonic generation (SHG) and three parameters for the transmittance of the fundamental laser beam. Using these parameters, it is possible to recover the Jones tensor elements of the sample at video rate. Video rate imaging is enabled by performing synchronous digitization (SD), in which a PCIe digital oscilloscope card is synchronized to the laser (the laser is the master clock.) Fast polarization modulation was achieved by modulating an electro-optic modulator synchronously with the laser and digitizer, with a simple sine-wave at 1/10th the period of the laser, producing a repeating pattern of 10 polarization states. This approach was validated using Z-cut quartz, and NOSE microscopy was performed for micro-crystals of naproxen.

Synchronous-digitization for video rate polarization modulated beam scanning second harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Sullivan, Shane Z.; DeWalt, Emma L.; Schmitt, Paul D.; Muir, Ryan D.; Simpson, Garth J.

2015-03-01

Fast beam-scanning non-linear optical microscopy, coupled with fast (8 MHz) polarization modulation and analytical modeling have enabled simultaneous nonlinear optical Stokes ellipsometry (NOSE) and linear Stokes ellipsometry imaging at video rate (15 Hz). NOSE enables recovery of the complex-valued Jones tensor that describes the polarization-dependent observables, in contrast to polarimetry, in which the polarization stated of the exciting beam is recorded. Each data acquisition consists of 30 images (10 for each detector, with three detectors operating in parallel), each of which corresponds to polarization-dependent results. Processing of this image set by linear fitting contracts down each set of 10 images to a set of 5 parameters for each detector in second harmonic generation (SHG) and three parameters for the transmittance of the fundamental laser beam. Using these parameters, it is possible to recover the Jones tensor elements of the sample at video rate. Video rate imaging is enabled by performing synchronous digitization (SD), in which a PCIe digital oscilloscope card is synchronized to the laser (the laser is the master clock.) Fast polarization modulation was achieved by modulating an electro-optic modulator synchronously with the laser and digitizer, with a simple sine-wave at 1/10th the period of the laser, producing a repeating pattern of 10 polarization states. This approach was validated using Z-cut quartz, and NOSE microscopy was performed for micro-crystals of naproxen.

Automated biphasic morphological assessment of hepatitis B-related liver fibrosis using second harmonic generation microscopy

PubMed Central

Wang, Tong-Hong; Chen, Tse-Ching; Teng, Xiao; Liang, Kung-Hao; Yeh, Chau-Ting

2015-01-01

Liver fibrosis assessment by biopsy and conventional staining scores is based on histopathological criteria. Variations in sample preparation and the use of semi-quantitative histopathological methods commonly result in discrepancies between medical centers. Thus, minor changes in liver fibrosis might be overlooked in multi-center clinical trials, leading to statistically non-significant data. Here, we developed a computer-assisted, fully automated, staining-free method for hepatitis B-related liver fibrosis assessment. In total, 175 liver biopsies were divided into training (n = 105) and verification (n = 70) cohorts. Collagen was observed using second harmonic generation (SHG) microscopy without prior staining, and hepatocyte morphology was recorded using two-photon excitation fluorescence (TPEF) microscopy. The training cohort was utilized to establish a quantification algorithm. Eleven of 19 computer-recognizable SHG/TPEF microscopic morphological features were significantly correlated with the ISHAK fibrosis stages (P < 0.001). A biphasic scoring method was applied, combining support vector machine and multivariate generalized linear models to assess the early and late stages of fibrosis, respectively, based on these parameters. The verification cohort was used to verify the scoring method, and the area under the receiver operating characteristic curve was >0.82 for liver cirrhosis detection. Since no subjective gradings are needed, interobserver discrepancies could be avoided using this fully automated method. PMID:26260921

Texture analysis applied to second harmonic generation image data for ovarian cancer classification

NASA Astrophysics Data System (ADS)

Wen, Bruce L.; Brewer, Molly A.; Nadiarnykh, Oleg; Hocker, James; Singh, Vikas; Mackie, Thomas R.; Campagnola, Paul J.

2014-09-01

Remodeling of the extracellular matrix has been implicated in ovarian cancer. To quantitate the remodeling, we implement a form of texture analysis to delineate the collagen fibrillar morphology observed in second harmonic generation microscopy images of human normal and high grade malignant ovarian tissues. In the learning stage, a dictionary of "textons"-frequently occurring texture features that are identified by measuring the image response to a filter bank of various shapes, sizes, and orientations-is created. By calculating a representative model based on the texton distribution for each tissue type using a training set of respective second harmonic generation images, we then perform classification between images of normal and high grade malignant ovarian tissues. By optimizing the number of textons and nearest neighbors, we achieved classification accuracy up to 97% based on the area under receiver operating characteristic curves (true positives versus false positives). The local analysis algorithm is a more general method to probe rapidly changing fibrillar morphologies than global analyses such as FFT. It is also more versatile than other texture approaches as the filter bank can be highly tailored to specific applications (e.g., different disease states) by creating customized libraries based on common image features.

Femtosecond laser nanosurgery of sub-cellular structures in HeLa cells by employing Third Harmonic Generation imaging modality as diagnostic tool.

PubMed

Tserevelakis, George J; Psycharakis, Stylianos; Resan, Bojan; Brunner, Felix; Gavgiotaki, Evagelia; Weingarten, Kurt; Filippidis, George

2012-02-01

Femtosecond laser assisted nanosurgery of microscopic biological specimens is a relatively new technique which allows the selective disruption of sub-cellular structures without causing any undesirable damage to the surrounding regions. The targeted structures have to be stained in order to be clearly visualized for the nanosurgery procedure. However, the validation of the final nanosurgery result is difficult, since the targeted structure could be simply photobleached rather than selectively destroyed. This fact comprises a main drawback of this technique. In our study we employed a multimodal system which integrates non-linear imaging modalities with nanosurgery capabilities, for the selective disruption of sub-cellular structures in HeLa cancer cells. Third Harmonic Generation (THG) imaging modality was used as a tool for the identification of structures that were subjected to nanosurgery experiments. No staining of the biological samples was required, since THG is an intrinsic property of matter. Furthermore, cells' viability after nanosurgery processing was verified via Two Photon Excitation Fluorescence (TPEF) measurements. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Quasiclassical Theory on Third-Harmonic Generation in Conventional Superconductors with Paramagnetic Impurities

NASA Astrophysics Data System (ADS)

Jujo, Takanobu

2018-02-01

We investigate the third-harmonic generation (THG) of s-wave superconductors under microwave pulse irradiation. We consider the effect of paramagnetic impurities on the THG intensity of dirty superconductors. The nonlinear response function is calculated using the method of the quasiclassical Green function. It is shown that the amplitude mode is included as the vertex correction and makes a predominant contribution to the THG intensity. When the effect of paramagnetic impurities is weak, the THG intensity shows a peak at the temperature at which the superconducting gap is about the same as the frequency of the incident pulse, similarly to in experiments. As the effect of paramagnetic impurities is strengthened, the peak of the THG intensity disappears. This indicates that time-reversal symmetry breaking due to paramagnetic impurities eliminates the well-defined amplitude mode. The result of our calculation shows that the existence of the amplitude mode can be confirmed through the THG intensity. The result of a semiquantitative calculation is in good agreement with the experimental result, and it also shows that the diamagnetic term is negligible.

Octave-spanning mid-infrared pulses by plasma generation in air pumped with an Yb:KGW source

PubMed Central

Huang, Jinqing; Parobek, Alexander; Ganim, Ziad

2016-01-01

Femtosecond mid-infrared (IR) supercontinuum generation in gas media provides a broadband source suited for time-domain spectroscopies and microscopies. This technology has largely utilized <100 fs Ti:sapphire pump lasers. In this Letter, we describe the first plasma generation mid-IR source based on a 1030 nm, 171 fs Yb:KGW laser system; when its first three harmonics are focused in air, a conical mode supercontinuum is generated that spans <1000 to 2700 cm−1 with a 190 pJ pulse energy and 0.5% RMS stability. PMID:27805634

Nonlinear optical behavior of DNA-functionalized gold nanoparticles

NASA Astrophysics Data System (ADS)

Kulyk, B.; Krupka, O.; Smokal, V.; Figà, V.; Czaplicki, R.; Sahraoui, B.

2018-03-01

The third-order nonlinear optical properties of gold nanoparticles embedded in the DNA-based composites were investigated by means of the third harmonic generation. With this purpose, the thin films comprising DNA-based complexes and Au nanoparticles were spin-deposited on glass substrate and their optical and nonlinear optical features were studied using the Maker-fringe technique at a laser fundamental wavelength of 1064 nm. The values of the third-order susceptibility χ (3)(- 3ω; ω, ω, ω) of the composite films based on DNA complex doped with 5 wt% of N-ethyl-N-(2-hydroxyethyl)-4-(4-nitrophenylazo)aniline were found to be significantly higher than those for pure composite films. Meanwhile, the presence of Au nanoparticles noticeable decreases the third-order nonlinear response of DNA-based composite mainly due to the enhanced absorption and scattering of laser and generated beam, respectively.

Characterization of muscle contraction with second harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Prent, Nicole

Muscle cells have the ability to change length and generate force due to orchestrated action of myosin nanomotors that cause sliding of actin filaments along myosin filaments in the sarcomeres, the fundamental contractile units, of myocytes. The correlated action of hundreds of sarcomeres is needed to produce the myocyte contractions. This study probes the molecular structure of the myofilaments and investigates the movement correlations between sarcomeres during contraction. In this study, second harmonic generation (SHG) microscopy is employed for imaging striated myocytes. Myosin filaments in striated myocytes inherently have a nonzero second-order susceptibility, [special characters omitted] and therefore generate efficient SHG. Employing polarization-in polarization-out (PIPO) SHG microscopy allows for the accurate determination of the characteristic ratio, [special characters omitted] in birefringent myocytes, which describes the structure of the myosin filament. Analysis shows that the b value at the centre of the myosin filament, where the nonlinear dipoles are better aligned, is slightly lower than the value at the edges of the filament, where there is more disorder in orientation of the nonlinear dipoles from the myosin heads. Forced stretching of myocytes resulted in an SHG intensity increase with the elongation of the sarcomere. SHG microscopy captured individual sarcomeres during contraction, allowing for the measurement of sarcomere length (SL) and SHG intensity (SI) fluctuations. The fluctuations also revealed higher SHG intensity in elongated sarcomeres. The sarcomere synchronization model (SSM) for contracting and quiescent myocytes was developed, and experimentally verified for three cases (isolated cardiomyocyte, embryonic chicken cardiomyocyte, and larva myocyte). During contraction, the action of SLs and SIs between neighbouring sarcomeres partially correlated, whereas in quiescent myocytes the SLs show an anti-correlation and the SIs have no correlations. The characteristic correlation coefficients and amplitudes were obtained for each case, allowing for the characterization of the synchronization of sarcomere movement during muscle contraction. These investigations constitute the basis for studying the structure and physiology of muscle cell contractions with polarization SHG microscopy. Live dynamic imaging of myocytes is applicable for contractility research, drug discovery, and as a diagnostic tool for monitoring muscular diseases.

Structure of polarization singularities of a light beam at triple frequency generated in isotropic medium by singularly polarized beam.

PubMed

Grigoriev, K S; Ryzhikov, P S; Cherepetskaya, E B; Makarov, V A

2017-10-16

The components of electric field of the third harmonic beam, generated in isotropic medium with cubic nonlinearity by a monochromatic light beam carrying polarization singularity of an arbitrary type, are found analytically. The relation between C-points characteristics in the fundamental and signal beams are determined, as well as the impact of the phase mismatch on the shape of the C-lines.

Imaging Collagen in Scar Tissue: Developments in Second Harmonic Generation Microscopy for Biomedical Applications

PubMed Central

Mostaço-Guidolin, Leila; Rosin, Nicole L.; Hackett, Tillie-Louise

2017-01-01

The ability to respond to injury with tissue repair is a fundamental property of all multicellular organisms. The extracellular matrix (ECM), composed of fibrillar collagens as well as a number of other components is dis-regulated during repair in many organs. In many tissues, scaring results when the balance is lost between ECM synthesis and degradation. Investigating what disrupts this balance and what effect this can have on tissue function remains an active area of research. Recent advances in the imaging of fibrillar collagen using second harmonic generation (SHG) imaging have proven useful in enhancing our understanding of the supramolecular changes that occur during scar formation and disease progression. Here, we review the physical properties of SHG, and the current nonlinear optical microscopy imaging (NLOM) systems that are used for SHG imaging. We provide an extensive review of studies that have used SHG in skin, lung, cardiovascular, tendon and ligaments, and eye tissue to understand alterations in fibrillar collagens in scar tissue. Lastly, we review the current methods of image analysis that are used to extract important information about the role of fibrillar collagens in scar formation. PMID:28809791

Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Sun, Tzu-Lin; Liu, Yuan; Sung, Ming-Chin; Chen, Hsiao-Ching; Yang, Chun-Hui; Hovhannisyan, Vladimir; Lin, Wei-Chou; Jeng, Yung-Ming; Chen, Wei-Liang; Chiou, Ling-Ling; Huang, Guan-Tarn; Kim, Ki-Hean; So, Peter T. C.; Chen, Yang-Fang; Lee, Hsuan-Shu; Dong, Chen-Yuan

2010-05-01

Conventionally, liver fibrosis is diagnosed using histopathological techniques. The traditional method is time-consuming in that the specimen preparation procedure requires sample fixation, slicing, and labeling. Our goal is to apply multiphoton microscopy to efficiently image and quantitatively analyze liver fibrosis specimens bypassing steps required in histological preparation. In this work, the combined imaging modality of multiphoton autofluorescence (MAF) and second-harmonic generation (SHG) was used for the qualitative imaging of liver fibrosis of different METAVIR grades under label-free, ex vivo conditions. We found that while MAF is effective in identifying cellular architecture in the liver specimens, it is the spectrally distinct SHG signal that allows the characterization of the extent of fibrosis. We found that qualitative SHG imaging can be used for the effective identification of the associated features of liver fibrosis specimens graded METAVIR 0 to 4. In addition, we attempted to associate quantitative SHG signal to the different METAVIR grades and found that an objective determination of the extent of disease progression can be made. Our approach demonstrates the potential of using multiphoton imaging in rapid classification of ex vivo liver fibrosis in the clinical setting and investigation of liver fibrosis-associated physiopathology in animal models in vivo.

Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy.

PubMed

Sun, Tzu-Lin; Liu, Yuan; Sung, Ming-Chin; Chen, Hsiao-Ching; Yang, Chun-Hui; Hovhannisyan, Vladimir; Lin, Wei-Chou; Jeng, Yung-Ming; Chen, Wei-Liang; Chiou, Ling-Ling; Huang, Guan-Tarn; Kim, Ki-Hean; So, Peter T C; Chen, Yang-Fang; Lee, Hsuan-Shu; Dong, Chen-Yuan

2010-01-01

Conventionally, liver fibrosis is diagnosed using histopathological techniques. The traditional method is time-consuming in that the specimen preparation procedure requires sample fixation, slicing, and labeling. Our goal is to apply multiphoton microscopy to efficiently image and quantitatively analyze liver fibrosis specimens bypassing steps required in histological preparation. In this work, the combined imaging modality of multiphoton autofluorescence (MAF) and second-harmonic generation (SHG) was used for the qualitative imaging of liver fibrosis of different METAVIR grades under label-free, ex vivo conditions. We found that while MAF is effective in identifying cellular architecture in the liver specimens, it is the spectrally distinct SHG signal that allows the characterization of the extent of fibrosis. We found that qualitative SHG imaging can be used for the effective identification of the associated features of liver fibrosis specimens graded METAVIR 0 to 4. In addition, we attempted to associate quantitative SHG signal to the different METAVIR grades and found that an objective determination of the extent of disease progression can be made. Our approach demonstrates the potential of using multiphoton imaging in rapid classification of ex vivo liver fibrosis in the clinical setting and investigation of liver fibrosis-associated physiopathology in animal models in vivo.

Application of Multiphoton Microscopy in Dermatological Studies: a Mini-Review

PubMed Central

Yew, Elijah; Rowlands, Christopher

2014-01-01

This review summarizes the historical and more recent developments of multiphoton microscopy, as applied to dermatology. Multiphoton microscopy offers several advantages over competing microscopy techniques: there is an inherent axial sectioning, penetration depths that compete well with confocal microscopy on account of the use of near-infrared light, and many two-photon contrast mechanisms, such as second-harmonic generation, have no analogue in one-photon microscopy. While the penetration depths of photons into tissue are typically limited on the order of hundreds of microns, this is of less concern in dermatology, as the skin is thin and readily accessible. As a result, multiphoton microscopy in dermatology has generated a great deal of interest, much of which is summarized here. The review covers the interaction of light and tissue, as well as the various considerations that must be made when designing an instrument. The state of multiphoton microscopy in imaging skin cancer and various other diseases is also discussed, along with the investigation of aging and regeneration phenomena, and finally, the use of multiphoton microscopy to analyze the transdermal transport of drugs, cosmetics and other agents is summarized. The review concludes with a look at potential future research directions, especially those that are necessary to push these techniques into widespread clinical acceptance. PMID:25075226

Advanced Methods in Fluorescence Microscopy

PubMed Central

Fritzky, Luke; Lagunoff, David

2013-01-01

It requires a good deal of will power to resist hyperbole in considering the advances that have been achieved in fluorescence microscopy in the last 25 years. Our effort has been to survey the modalities of microscopic fluorescence imaging available to cell biologists and perhaps useful for diagnostic pathologists. The gamut extends from established confocal laser scanning through multiphoton and TIRF to the emerging technologies of super-resolution microscopy that breech the Abbé limit of resolution. Also considered are the recent innovations in structured and light sheet illumination, the use of FRET and molecular beacons that exploit specific characteristics of designer fluorescent proteins, fluorescence speckles, and second harmonic generation for native anisometric structures like collagen, microtubules and sarcomeres. PMID:23271142

Advanced methods in fluorescence microscopy.

PubMed

Fritzky, Luke; Lagunoff, David

2013-01-01

It requires a good deal of will power to resist hyperbole in considering the advances that have been achieved in fluorescence microscopy in the last 25 years. Our effort has been to survey the modalities of microscopic fluorescence imaging available to cell biologists and perhaps useful for diagnostic pathologists. The gamut extends from established confocal laser scanning through multiphoton and TIRF to the emerging technologies of super-resolution microscopy that breech the Abbe limit of resolution. Also considered are the recent innovations in structured and light sheet illumination, the use of FRET and molecular beacons that exploit specific characteristics of designer fluorescent proteins, fluorescence speckles, and second harmonic generation for native anisometric structures like collagen, microtubules and sarcomeres.

Advanced methods in fluorescence microscopy.

PubMed

Fritzky, Luke; Lagunoff, David

2013-01-01

It requires a good deal of will power to resist hyperbole in considering the advances that have been achieved in fluorescence microscopy in the last 25 years. Our effort has been to survey the modalities of microscopic fluorescence imaging available to cell biologists and perhaps useful for diagnostic pathologists. The gamut extends from established confocal laser scanning through multiphoton and TIRF to the emerging technologies of super-resolution microscopy that breech the Abbé limit of resolution. Also considered are the recent innovations in structured and light sheet illumination, the use of FRET and molecular beacons that exploit specific characteristics of designer fluorescent proteins, fluorescence speckles, and second harmonic generation for native anisometric structures like collagen, microtubules and sarcomeres.

Theoretical Study on the 1.185-THz Third Harmonic Gyrotron

NASA Astrophysics Data System (ADS)

Dumbrajs, O.; Idehara, T.

2018-02-01

We discuss how the existing University of Fukui (FIR UF) second harmonic double-beam gyrotron with the operating frequency 0.79 THz can be adopted for operation at the third harmonic. The new gyrotron will operate at the frequency 1.185 THz and will significantly increase the frequency of the dynamic nuclear polarization-nuclear magnetic resonance (DNP-NMR) spectrometer. This will allow one to study new bio-molecules.A special attention is payed to the mode competition between the operating {TE}_{3,11}+ mode at the third harmonic and the parasitic modes at the second and fundamental harmonics. The operating parameters of the modified gyrotron are U = 20 kV, α = 1.3, I = 0.35 A, and B = 14.60 T and the expected output power about 100W.

Third-order optical conductivity of an electron fluid

NASA Astrophysics Data System (ADS)

Sun, Zhiyuan; Basov, D. N.; Fogler, M. M.

2018-02-01

We derive the nonlinear optical conductivity of an isotropic electron fluid at frequencies below the interparticle collision rate. In this regime, governed by hydrodynamics, the conductivity acquires a universal form at any temperature, chemical potential, and spatial dimension. We show that the nonlinear response of the fluid to a uniform field is dominated by the third-order conductivity tensor σ(3 ) whose magnitude and temperature dependence differ qualitatively from those in the conventional kinetic regime of higher frequencies. We obtain explicit formulas for σ(3 ) for Dirac materials such as graphene and Weyl semimetals. We make predictions for the third-harmonic generation, renormalization of the collective-mode spectrum, and the third-order circular magnetic birefringence experiments.

Intercalating dyes for enhanced contrast in second-harmonic generation imaging of protein crystals

PubMed Central

Newman, Justin A.; Scarborough, Nicole M.; Pogranichniy, Nicholas R.; Shrestha, Rashmi K.; Closser, Richard G.; Das, Chittaranjan; Simpson, Garth J.

2015-01-01

The second-harmonic generation (SHG) activity of protein crystals was found to be enhanced by up to ∼1000-fold by the intercalation of SHG phores within the crystal lattice. Unlike the intercalation of fluorophores, the SHG phores produced no significant background SHG from solvated dye or from dye intercalated into amorphous aggregates. The polarization-dependent SHG is consistent with the chromophores adopting the symmetry of the crystal lattice. In addition, the degree of enhancement for different symmetries of dyes is consistent with theoretical predictions based on the molecular nonlinear optical response. Kinetics studies indicate that intercalation arises over a timeframe of several minutes in lysozyme, with detectable enhancements within seconds. These results provide a potential means to increase the overall diversity of protein crystals and crystal sizes amenable to characterization by SHG microscopy. PMID:26143918

Application of Fourier transform-second-harmonic generation imaging to the rat cervix.

PubMed

Lau, T Y; Sangha, H K; Chien, E K; McFarlin, B L; Wagoner Johnson, A J; Toussaint, K C

2013-07-01

We present the application of Fourier transform-second-harmonic generation (FT-SHG) imaging to evaluate the arrangement of collagen fibers in five nonpregnant rat cervices. Tissue slices from the mid-cervix and near the external orifice of the cervix were analyzed in both two-dimensions (2D) and three-dimensions (3D). We validate that the cervical microstructure can be quantitatively assessed in three dimensions using FT-SHG imaging and observe collagen fibers oriented both in and out-of-plane in the outermost and the innermost layers, which cannot be observed using 2D FT-SHG analysis alone. This approach has the potential to be a clinically applicable method for measuring progressive changes in collagen organization during cervical remodeling in humans. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

Millimeter-wave active probe

DOEpatents

Majidi-Ahy, Gholamreza; Bloom, David M.

1991-01-01

A millimeter-wave active probe for use in injecting signals with frequencies above 50GHz to millimeter-wave and ultrafast devices and integrated circuits including a substrate upon which a frequency multiplier consisting of filter sections and impedance matching sections are fabricated in uniplanar transmission line format. A coaxial input and uniplanar 50 ohm transmission line couple an approximately 20 GHz input signal to a low pass filter which rolls off at approximately 25 GHz. An input impedance matching section couples the energy from the low pass filter to a pair of matched, antiparallel beam lead diodes. These diodes generate odd-numberd harmonics which are coupled out of the diodes by an output impedance matching network and bandpass filter which suppresses the fundamental and third harmonics and selects the fifth harmonic for presentation at an output.

Temperature dependence of acoustic harmonics generated by nonlinear ultrasound beam propagation in ex vivo tissue and tissue-mimicking phantoms.

PubMed

Maraghechi, Borna; Kolios, Michael C; Tavakkoli, Jahan

2015-01-01

Hyperthermia is a cancer treatment technique that could be delivered as a stand-alone modality or in conjunction with chemotherapy or radiation therapy. Noninvasive and real-time temperature monitoring of the heated tissue improves the efficacy and safety of the treatment. A temperature-sensitive acoustic parameter is required for ultrasound-based thermometry. In this paper the amplitude and the energy of the acoustic harmonics of the ultrasound backscattered signal are proposed as suitable parameters for noninvasive ultrasound thermometry. A commercial high frequency ultrasound imaging system was used to generate and detect acoustic harmonics in tissue-mimicking gel phantoms and ex vivo bovine muscle tissues. The pressure amplitude and the energy content of the backscattered fundamental frequency (p1 and E1), the second (p2 and E2) and the third (p3 and E3) harmonics were detected in pulse-echo mode. Temperature was increased from 26° to 46 °C uniformly through both samples. The amplitude and the energy content of the harmonics and their ratio were measured and analysed as a function of temperature. The average p1, p2 and p3 increased by 69%, 100% and 283%, respectively as the temperature was elevated from 26° to 46 °C in tissue samples. In the same experiment the average E1, E2 and E3 increased by 163%, 281% and 2257%, respectively. A similar trend was observed in tissue-mimicking gel phantoms. The findings suggest that the harmonics generated due to nonlinear ultrasound beam propagation are highly sensitive to temperature and could potentially be used for noninvasive ultrasound tissue thermometry.

Modelling third harmonic ion cyclotron acceleration of deuterium beams for JET fusion product studies experiments

NASA Astrophysics Data System (ADS)

Schneider, M.; Johnson, T.; Dumont, R.; Eriksson, J.; Eriksson, L.-G.; Giacomelli, L.; Girardo, J.-B.; Hellsten, T.; Khilkevitch, E.; Kiptily, V. G.; Koskela, T.; Mantsinen, M.; Nocente, M.; Salewski, M.; Sharapov, S. E.; Shevelev, A. E.; Contributors, JET

2016-11-01

Recent JET experiments have been dedicated to the studies of fusion reactions between deuterium (D) and Helium-3 (3He) ions using neutral beam injection (NBI) in synergy with third harmonic ion cyclotron radio-frequency heating (ICRH) of the beam. This scenario generates a fast ion deuterium tail enhancing DD and D3He fusion reactions. Modelling and measuring the fast deuterium tail accurately is essential for quantifying the fusion products. This paper presents the modelling of the D distribution function resulting from the NBI+ICRF heating scheme, reinforced by a comparison with dedicated JET fast ion diagnostics, showing an overall good agreement. Finally, a sawtooth activity for these experiments has been observed and interpreted using SPOT/RFOF simulations in the framework of Porcelli’s theoretical model, where NBI+ICRH accelerated ions are found to have a strong stabilizing effect, leading to monster sawteeth.

Superharmonic imaging with chirp coded excitation: filtering spectrally overlapped harmonics.

PubMed

Harput, Sevan; McLaughlan, James; Cowell, David M J; Freear, Steven

2014-11-01

Superharmonic imaging improves the spatial resolution by using the higher order harmonics generated in tissue. The superharmonic component is formed by combining the third, fourth, and fifth harmonics, which have low energy content and therefore poor SNR. This study uses coded excitation to increase the excitation energy. The SNR improvement is achieved on the receiver side by performing pulse compression with harmonic matched filters. The use of coded signals also introduces new filtering capabilities that are not possible with pulsed excitation. This is especially important when using wideband signals. For narrowband signals, the spectral boundaries of the harmonics are clearly separated and thus easy to filter; however, the available imaging bandwidth is underused. Wideband excitation is preferable for harmonic imaging applications to preserve axial resolution, but it generates spectrally overlapping harmonics that are not possible to filter in time and frequency domains. After pulse compression, this overlap increases the range side lobes, which appear as imaging artifacts and reduce the Bmode image quality. In this study, the isolation of higher order harmonics was achieved in another domain by using the fan chirp transform (FChT). To show the effect of excitation bandwidth in superharmonic imaging, measurements were performed by using linear frequency modulated chirp excitation with varying bandwidths of 10% to 50%. Superharmonic imaging was performed on a wire phantom using a wideband chirp excitation. Results were presented with and without applying the FChT filtering technique by comparing the spatial resolution and side lobe levels. Wideband excitation signals achieved a better resolution as expected, however range side lobes as high as -23 dB were observed for the superharmonic component of chirp excitation with 50% fractional bandwidth. The proposed filtering technique achieved >50 dB range side lobe suppression and improved the image quality without affecting the axial resolution.

Experimental observation of breathing solitons and a third harmonic in a tapered photonic crystal fiber

NASA Astrophysics Data System (ADS)

Wang, F.; Yao, C. F.; Li, C. Z.; Jia, Z. X.; Li, Q.; Wu, C. F.; Ohishi, Y.; Qin, W. P.; Qin, G. S.

2018-02-01

We report the experimental observation of breathing solitons and a third harmonic in a tapered fluorotellurite photonic crystal fiber (PCF) pumped by a 1560 nm femtosecond fiber laser. The PCF has a core diameter that varies continuously along its length, resulting in a zero-dispersion wavelength that moves from 1325 nm to 906 nm over the transition region. By finely controlling the dispersion map of the tapered PCF and increasing the order of the optical solitons, their breathing behavior is observed in the frequency domain and the number of breaths goes up to 9. Furthermore, the breathing behavior of the optical soliton is transferred to the third harmonic through inter-modal phase-matched processes in the tapered PCF, and the third harmonic also breathes with an increase in the pump power.

A hybrid ruthenium alkynyl/zinc porphyrin "Cross Fourchée" with large cubic NLO properties.

PubMed

Merhi, Areej; Grelaud, Guillaume; Green, Katy A; Minh, Ngo Hoang; Reynolds, Michael; Ledoux, Isabelle; Barlow, Adam; Wang, Genmiao; Cifuentes, Marie P; Humphrey, Mark G; Paul, Frédéric; Paul-Roth, Christine O

2015-05-07

A new Zn(ii) porphyrin-cored ruthenium alkynyl dendrimer (2) containing twelve Ru(κ(2)-dppe)2 bis-alkynyl fragments has been prepared in two steps from 5,10,15,20-tetra(4-ethynylphenyl)porphyrinatozinc(ii) and shown to be highly active for third-harmonic generation (THG) at 1907 nm.

In vivo multimodal nonlinear optical imaging of mucosal tissue

NASA Astrophysics Data System (ADS)

Sun, Ju; Shilagard, Tuya; Bell, Brent; Motamedi, Massoud; Vargas, Gracie

2004-05-01

We present a multimodal nonlinear imaging approach to elucidate microstructures and spectroscopic features of oral mucosa and submucosa in vivo. The hamster buccal pouch was imaged using 3-D high resolution multiphoton and second harmonic generation microscopy. The multimodal imaging approach enables colocalization and differentiation of prominent known spectroscopic and structural features such as keratin, epithelial cells, and submucosal collagen at various depths in tissue. Visualization of cellular morphology and epithelial thickness are in excellent agreement with histological observations. These results suggest that multimodal nonlinear optical microscopy can be an effective tool for studying the physiology and pathology of mucosal tissue.

Characterization of corneal damage from Pseudomonas aeruginosa infection by the use of multiphoton microscopy

NASA Astrophysics Data System (ADS)

Chang, Yu-Lin; Chen, Wei-Liang; Lo, Wen; Chen, Shean-Jen; Tan, Hsin-Yuan; Dong, Chen-Yuan

2010-11-01

Using multiphoton autofluorescence (MAF) and second harmonic generation (SHG) microscopy, we investigate the morphology and the structure of the corneal epithelium and stroma collagen of bovine cornea following injection of Pseudomonas aeruginosa. We found that corneal epithelial cells are damaged and stromal collagen becoming increasingly autofluorescent with time. We also characterized infected cornea cultured for 0, 6, 12, and 24 h by quantitative ratiometric MAF to SHG index (MAFSI) analysis. MAFSI results show that the destruction of the stromal collagen corresponds to a decrease in SHG intensity and increase of MAF signal with time.

Third-harmonic generation and scattering in combustion flames using a femtosecond laser filament.

PubMed

Zang, Hong-Wei; Li, He-Long; Su, Yue; Fu, Yao; Hou, Meng-Yao; Baltuška, Andrius; Yamanouchi, Kaoru; Xu, Huailiang

2018-02-01

Coherent radiation in the ultraviolent (UV) range has high potential applicability to the diagnosis of the formation processes of soot in combustion because of the high scattering efficiency in the UV wavelength region, even though the UV light is lost largely by the absorption within the combustion flames. We show that the third harmonic (TH) of a Ti:sapphire 800 nm femtosecond laser is generated in a laser-induced filament in a combustion flame and that the conversion efficiency of the TH varies sensitively by the ellipticity of the driver laser pulse but does not vary so much by the choice of alkanol species introduced as fuel for the combustion flames. We also find that the TH recorded from the side direction of the filament is the Rayleigh scattering of the TH by soot nanoparticles within the flame and that the intensity of the TH varies depending on the fuel species as well as on the position of the laser filament within the flame. Our results show that a remote and in situ measurement of distributions of soot nanoparticles in a combustion flame can be achieved by Rayleigh scattering spectroscopy of the TH generated by a femtosecond-laser-induced filament in the combustion flame.

Cellulose amorphization by swelling in ionic liquid/water mixtures: a combined macroscopic and second-harmonic microscopy study.

PubMed

Glas, Daan; Paesen, Rik; Depuydt, Daphne; Binnemans, Koen; Ameloot, Marcel; De Vos, Dirk E; Ameloot, Rob

2015-01-01

Amorphization of cellulose by swelling in ionic liquid (IL)/water mixtures at room temperature is a suitable alternative to the dissolution-precipitation pretreatment known to facilitate enzymatic digestion. When soaking microcrystalline cellulose in the IL 1-ethyl-3-methylimidazolium acetate containing 20 wt % water, the crystallinity of the cellulose sample is strongly reduced. As less than 4 % of the cellulose dissolves in this mixture, this swelling method makes a precipitation step and subsequent energy-intensive IL purification redundant. Second-harmonic generation (SHG) microscopy is used as a structure-sensitive technique for in situ monitoring of the changes in cellulose crystallinity. Combined optical and SHG observations confirm that in the pure IL complete dissolution takes place, while swelling without dissolution in the optimal IL/water mixture yields a solid cellulose with a significantly reduced crystallinity in a single step. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A robust collagen scoring method for human liver fibrosis by second harmonic microscopy.

PubMed

Guilbert, Thomas; Odin, Christophe; Le Grand, Yann; Gailhouste, Luc; Turlin, Bruno; Ezan, Frédérick; Désille, Yoann; Baffet, Georges; Guyader, Dominique

2010-12-06

Second Harmonic Generation (SHG) microscopy offers the opportunity to image collagen of type I without staining. We recently showed that a simple scoring method, based on SHG images of histological human liver biopsies, correlates well with the Metavir assessment of fibrosis level (Gailhouste et al., J. Hepatol., 2010). In this article, we present a detailed study of this new scoring method with two different objective lenses. By using measurements of the objectives point spread functions and of the photomultiplier gain, and a simple model of the SHG intensity, we show that our scoring method, applied to human liver biopsies, is robust to the objective's numerical aperture (NA) for low NA, the choice of the reference sample and laser power, and the spatial sampling rate. The simplicity and robustness of our collagen scoring method may open new opportunities in the quantification of collagen content in different organs, which is of main importance in providing diagnostic information and evaluation of therapeutic efficiency.

Optical Quantification of Harmonic Acoustic Radiation Force Excitation in a Tissue-Mimicking Phantom.

PubMed

Suomi, Visa; Edwards, David; Cleveland, Robin

2015-12-01

Optical tracking was used to characterize acoustic radiation force-induced displacements in a tissue-mimicking phantom. Amplitude-modulated 3.3-MHz ultrasound was used to induce acoustic radiation force in the phantom, which was embedded with 10-μm microspheres that were tracked using a microscope objective and high-speed camera. For sine and square amplitude modulation, the harmonic components of the fundamental and second and third harmonic frequencies were measured. The displacement amplitudes were found to increase linearly with acoustic radiation force up to 10 μm, with sine modulation having 19.5% lower peak-to-peak amplitude values than square modulation. Square modulation produced almost no second harmonic, but energy was present in the third harmonic. For the sine modulation, energy was present in the second harmonic and low energy in the third harmonic. A finite-element model was used to simulate the deformation and was both qualitatively and quantitatively in agreement with the measurements. Copyright © 2015 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

Towards improved NDE and SHM methodologies incorporating nonlinear structural features

NASA Astrophysics Data System (ADS)

Chillara, Vamshi Krishna

Ultrasound is widely employed in Nondestructive Evaluation (NDE) and Structural Health Monitoring (SHM) applications to detect and characterize damage/defects in materials. In particular, ultrasonic guided waves are considered a foremost candidate for in-situ monitoring applications. Conventional ultrasonic techniques rely on changes/discontinuities in linear elastic material properties, namely the Young's modulus and shear modulus to detect damage. On the other hand, nonlinear ultrasonic techniques that rely on micro-scale nonlinear material/structural behavior are proven to be sensitive to damage induced microstructural changes that precede macro-scale damage and are hence capable of early damage detection. The goal of this thesis is to investigate the capabilities of nonlinear guided waves --- a fusion of nonlinear ultrasonic techniques with the guided wave methodologies for early damage detection. To that end, the thesis focuses on two important aspects of the problem: 1. Wavemechanics - deals with ultrasonic guided wave propagation in nonlinear waveguides; 2. Micromechanics - deals with correlating ultrasonic response with micro-scale nonlinear material behavior. For the development of efficient NDE and SHM methodologies that incorporate nonlinear structural features, a detailed understanding of the above aspects is indispensable. In this thesis, the wavemechanics aspect of the problem is dealt with from both theoretical and numerical standpoints. A generalized theoretical framework is developed to study higher harmonic guided waves in plates. This was employed to study second harmonic guided waves in pipes using a large-radius asymptotic approximation. Second harmonic guided waves in plates are studied from a numerical standpoint. Theoretical predictions are validated and some key aspects of higher harmonic generation in waveguides are outlined. Finally, second harmonic guided waves in plates with inhomogeneous and localized nonlinearities are studied and some important aspects of guided wave mode selection are addressed. The other part of the work focused on developing a micromechanics based understanding of ultrasonic higher harmonic generation. Three important aspects of micro-scale material behavior, namely tension-compression asymmetry, shearnormal coupling and deformation induced asymmetry are identified and their role in ultrasonic higher harmonic generation is discussed. Tension-compression asymmetry is identified to cause second (even) harmonic generation in materials. Then, shearnormal coupling is identified to cause generation of secondary waves of different polarity than the primary waves. In addition, deformation induced anisotropy due to the presence of residual stress/strain and its contribution to ultrasonic higher harmonic generation is qualitatively discussed. Also, the tension-compression asymmetry in the material is quantified using an energy based measure. The above measure is employed to develop a homogenization based approach amenable to multi-scale analysis to correlate microstructure with ultrasonic higher harmonic generation. Finally, experimental investigations concerning third harmonic SH wave generation in plates are carried out and the effect of load and temperature changes on nonlinear ultrasonic measurements are discussed in the context of SHM. It was found that while nonlinear ultrasound is sensitive to micro-scale damage, the relative nonlinearity parameter may not always be the best measure to quantify the nonlinearity as it is subject to spurious effects from changes in environmental factors such as loads and temperature.

Fully automated muscle quality assessment by Gabor filtering of second harmonic generation images

NASA Astrophysics Data System (ADS)

Paesen, Rik; Smolders, Sophie; Vega, José Manolo de Hoyos; Eijnde, Bert O.; Hansen, Dominique; Ameloot, Marcel

2016-02-01

Although structural changes on the sarcomere level of skeletal muscle are known to occur due to various pathologies, rigorous studies of the reduced sarcomere quality remain scarce. This can possibly be explained by the lack of an objective tool for analyzing and comparing sarcomere images across biological conditions. Recent developments in second harmonic generation (SHG) microscopy and increasing insight into the interpretation of sarcomere SHG intensity profiles have made SHG microscopy a valuable tool to study microstructural properties of sarcomeres. Typically, sarcomere integrity is analyzed by fitting a set of manually selected, one-dimensional SHG intensity profiles with a supramolecular SHG model. To circumvent this tedious manual selection step, we developed a fully automated image analysis procedure to map the sarcomere disorder for the entire image at once. The algorithm relies on a single-frequency wavelet-based Gabor approach and includes a newly developed normalization procedure allowing for unambiguous data interpretation. The method was validated by showing the correlation between the sarcomere disorder, quantified by the M-band size obtained from manually selected profiles, and the normalized Gabor value ranging from 0 to 1 for decreasing disorder. Finally, to elucidate the applicability of our newly developed protocol, Gabor analysis was used to study the effect of experimental autoimmune encephalomyelitis on the sarcomere regularity. We believe that the technique developed in this work holds great promise for high-throughput, unbiased, and automated image analysis to study sarcomere integrity by SHG microscopy.

A junction-level optoelectronic characterization of etching-induced damage for third-generation HgCdTe infrared focal-plane array photodetectors

NASA Astrophysics Data System (ADS)

Wang, Peng; Wang, Yueming; Wu, Mingzai; Ye, Zhenhua

2018-06-01

Third-generation HgCdTe-based infrared focal plane arrays require high aspect ratio trenches with admissible etch induced damage at the surface and sidewalls for effectively isolating the pixels. In this paper, the high-density inductively coupled plasma enhanced reaction ion etching technique has been used for micro-mesa delineation of HgCdTe for third-generation infrared focal-plane array detectors. A nondestructive junction-level optoelectronic characterization method called laser beam induced current (LBIC) is used to evaluate the lateral junction extent of HgCdTe etch-induced damage scanning electron microscopy. It is found that the LBIC profiles exhibit evident double peaks and valleys phenomena. The lateral extent of etch induced mesa damage of ∼2.4 μm is obtained by comparing the LBIC profile and the scanning electron microscopy image of etched sample. This finding will guide us to nondestructively identify the distributions of the etching damages in large scale HgCdTe micro-mesa.

Efficient Third Harmonic Generation for Wind Lidar Applications

NASA Technical Reports Server (NTRS)

Mordaunt, David W.; Cheung, Eric C.; Ho, James G.; Palese, Stephen P.

1998-01-01

The characterization of atmospheric winds on a global basis is a key parameter required for accurate weather prediction. The use of a space based lidar system for remote measurement of wind speed would provide detailed and highly accurate data for future weather prediction models. This paper reports the demonstration of efficient third harmonic conversion of a 1 micrometer laser to provide an ultraviolet (UV) source suitable for a wind lidar system based on atmospheric molecular scattering. Although infrared based lidars using aerosol scattering have been demonstrated to provide accurate wind measurement, a UV based system using molecular or Rayleigh scattering will provide accurate global wind measurements, even in those areas of the atmosphere where the aerosol density is too low to yield good infrared backscatter signals. The overall objective of this work is to demonstrate the maturity of the laser technology and its suitability for a near term flight aboard the space shuttle. The laser source is based on diode-pumped solid-state laser technology which has been extensively demonstrated at TRW in a variety of programs and internal development efforts. The pump laser used for the third harmonic demonstration is a breadboard system, designated the Laser for Risk Reduction Experiments (LARRE), which has been operating regularly for over 5 years. The laser technology has been further refined in an engineering model designated as the Compact Advanced Pulsed Solid-State Laser (CAPSSL), in which the laser head was packaged into an 8 x 8 x 18 inch volume with a weight of approximately 61 pounds. The CAPSSL system is a ruggedized configuration suitable for typical military applications. The LARRE and CAPSSL systems are based on Nd:YAG with an output wavelength of 1064 nm. The current work proves the viability of converting the Nd:YAG fundamental to the third harmonic wavelength at 355 nm for use in a direct detection wind lidar based on atmospheric Rayleigh scattering.

Three-photon imaging of ovarian cancer

NASA Astrophysics Data System (ADS)

Barton, Jennifer K.; Amirsolaimani, Babak; Rice, Photini; Hatch, Kenneth; Kieu, Khanh

2016-02-01

Optical imaging methods have the potential to detect ovarian cancer at an early, curable stage. Optical imaging has the disadvantage that high resolution techniques require access to the tissue of interest, but miniature endoscopes that traverse the natural orifice of the reproductive tract, or access the ovaries and fallopian tubes through a small incision in the vagina wall, can provide a minimally-invasive solution. We have imaged both rodent and human ovaries and fallopian tubes with a variety of endoscope-compatible modalities. The recent development of fiber-coupled femtosecond lasers will enable endoscopic multiphoton microscopy (MPM). We demonstrated two- and three-photon excited fluorescence (2PEF, 3PEF), and second- and third-harmonic generation microscopy (SHG, THG) in human ovarian and fallopian tube tissue. A study was undertaken to understand the mechanisms of contrast in these images. Six patients (normal, cystadenoma, and ovarian adenocarcinoma) provided ovarian and fallopian tube biopsies. The tissue was imaged with three-dimensional optical coherence tomography, multiphoton microscopy, and frozen for histological sectioning. Tissue sections were stained with hematoxylin and eosin, Masson's trichrome, and Sudan black. Approximately 1 μm resolution images were obtained with an excitation source at 1550 nm. 2PEF signal was absent. SHG signal was mainly from collagen. 3PEF and THG signal came from a variety of sources, including a strong signal from fatty connective tissue and red blood cells. Adenocarcinoma was characterized by loss of SHG signal, whereas cystic abnormalities showed strong SHG. There was limited overlap of two- and three- photon signals, suggesting that three-photon imaging can provide additional information for early diagnosis of ovarian cancer.

Second Harmonic Generation Guided Raman Spectroscopy for Sensitive Detection of Polymorph Transitions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chowdhury, Azhad U.; Ye, Dong Hye; Song, Zhengtian

Second harmonic generation (SHG) was integrated with Raman spectroscopy for the analysis of pharmaceutical materials. Particulate formulations of clopidogrel bisulfate were prepared in two crystal forms (Form I and Form II). Image analysis approaches enable automated identification of particles by bright field imaging, followed by classification by SHG. Quantitative SHG microscopy enabled discrimination of crystal form on a per particle basis with 99.95% confidence in a total measurement time of ~10 ms per particle. Complementary measurements by Raman and synchrotron XRD are in excellent agreement with the classifications made by SHG, with measurement times of ~1 min and several secondsmore » per particle, respectively. Coupling these capabilities with at-line monitoring may enable real-time feedback for reaction monitoring during pharmaceutical production to favor the more bioavailable but metastable Form I with limits of detection in the ppm regime.« less

Character of skin on photo-thermal response and its regeneration process using second-harmonic generation microscopy.

PubMed

Wu, Shu-lian; Li, Hui; Zhang, Xiao-man; Chen, Wei R; Wang, Yun-Xia

2014-01-01

Quantitative characterization of skin collagen on photo-thermal response and its regeneration process is an important but difficult task. In this study, morphology and spectrum characteristics of collagen during photo-thermal response and its light-induced remodeling process were obtained by second-harmonic generation microscope in vivo. The texture feature of collagen orientation index and fractal dimension was extracted by image processing. The aim of this study is to detect the information hidden in skin texture during the process of photo-thermal response and its regeneration. The quantitative relations between injured collagen and texture feature were established for further analysis of the injured characteristics. Our results show that it is feasible to determine the main impacts of phototherapy on the skin. It is important to understand the process of collagen remodeling after photo-thermal injuries from texture feature.

Multimodal, multiphoton microscopy and image correlation analysis for characterizing corneal thermal damage

NASA Astrophysics Data System (ADS)

Lo, Wen; Chang, Yu-Lin; Liu, Jia-Shiu; Hseuh, Chiu-Mei; Hovhannisyan, Vladimir; Chen, Shean-Jen; Tan, Hsin-Yuan; Dong, Chen-Yuan

2009-09-01

We used the combination of multiphoton autofluorescence (MAF), forward second-harmonic generation (FWSHG), and backward second-harmonic generation (BWSHG) imaging for the qualitative and quantitative characterization of thermal damage of ex vivo bovine cornea. We attempt to characterize the structural alterations by qualitative MAF, FWSHG, and BWSHG imaging in the temperature range of 37 to 90°C. In addition to measuring the absolute changes in the three types of signals at the stromal surface, we also performed image correlation analysis between FWSHG and BWSHG and demonstrate that with increasing thermal damage, image correlation between FWSHG and BWSHG significantly increases. Our results show that while MAF and BWSHG intensities may be used as preliminary indicators of the extent of corneal thermal damage, the most sensitive measures are provided by the decay in FWSHG intensity and the convergence of FWSHG and BWSHG images.

Utilizing two-photon fluorescence and second harmonic generation microscopy to study human bone marrow mesenchymal stem cell morphogenesis in chitosan scaffold

NASA Astrophysics Data System (ADS)

Su, Ping-Jung; Huang, Chi-Hsiu; Huang, Yi-You; Lee, Hsuan-Sue; Dong, Chen-Yuan

2008-02-01

A major goal of tissue engineering is to cultivate the cartilage in vitro. One approach is to implant the human bone marrow mesenchymal stem cells into the three dimensional biocompatible and biodegradable material. Through the action of the chondrogenic factor TGF-β3, the stem cells can be induced to secrete collagen. In this study, mesenchymal stem cells are implanted on the chitosan scaffold and TGF-β3 was added to produce the cartilage tissue and TP autofluorescence and SHG microscopy was used to image the process of chondrogenesis. With additional development, multiphoton microscopy can be developed into an effective tool for evaluating the quality of tissue engineering products.

Quantitative second-harmonic generation imaging to detect osteogenesis imperfecta in human skin samples

NASA Astrophysics Data System (ADS)

Adur, J.; Ferreira, A. E.; D'Souza-Li, L.; Pelegati, V. B.; de Thomaz, A. A.; Almeida, D. B.; Baratti, M. O.; Carvalho, H. F.; Cesar, C. L.

2012-03-01

Osteogenesis Imperfecta (OI) is a genetic disorder that leads to bone fractures due to mutations in the Col1A1 or Col1A2 genes that affect the primary structure of the collagen I chain with the ultimate outcome in collagen I fibrils that are either reduced in quantity or abnormally organized in the whole body. A quick test screening of the patients would largely reduce the sample number to be studied by the time consuming molecular genetics techniques. For this reason an assessment of the human skin collagen structure by Second Harmonic Generation (SHG) can be used as a screening technique to speed up the correlation of genetics/phenotype/OI types understanding. In the present work we have used quantitative second harmonic generation (SHG) imaging microscopy to investigate the collagen matrix organization of the OI human skin samples comparing with normal control patients. By comparing fibril collagen distribution and spatial organization, we calculated the anisotropy and texture patterns of this structural protein. The analysis of the anisotropy was performed by means of the two-dimensional Discrete Fourier Transform and image pattern analysis with Gray-Level Co-occurrence Matrix (GLCM). From these results, we show that statistically different results are obtained for the normal and disease states of OI.

Monitoring the effect of mechanical stress on mesenchymal stem cell collagen production by multiphoton microscopy

NASA Astrophysics Data System (ADS)

Chen, Wei-Liang; Chang, Chia-Cheng; Chiou, Ling-Ling; Li, Tsung-Hsien; Liu, Yuan; Lee, Hsuan-Shu; Dong, Chen-Yuan

2008-02-01

Tissue engineering is emerging as a promising method for repairing damaged tissues. Due to cartilage's common wear and injury, in vitro production of cartilage replacements have been an active area of research. Finding the optimal condition for the generation of the collagen matrix is crucial in reproducing cartilages that closely match those found in human. Using multiphoton autofluorescence and second-harmonic generation (SHG) microscopy we monitored the effect of mechanical stress on mesenchymal stem cell collagen production. Bone marrow mesenchymal stem cells in the form of pellets were cultured and periodically placed under different mechanical stress by centrifugation over a period of four weeks. The differently stressed samples were imaged several times during the four week period, and the collagen production under different mechanical stress is characterized.

Structural distortion-induced magnetoelastic locking in Sr(2)IrO(4) revealed through nonlinear optical harmonic generation.

PubMed

Torchinsky, D H; Chu, H; Zhao, L; Perkins, N B; Sizyuk, Y; Qi, T; Cao, G; Hsieh, D

2015-03-06

We report a global structural distortion in Sr_{2}IrO_{4} using spatially resolved optical second and third harmonic generation rotational anisotropy measurements. A symmetry lowering from an I4_{1}/acd to I4_{1}/a space group is observed both above and below the Néel temperature that arises from a staggered tetragonal distortion of the oxygen octahedra. By studying an effective superexchange Hamiltonian that accounts for this lowered symmetry, we find that perfect locking between the octahedral rotation and magnetic moment canting angles can persist even in the presence of large noncubic local distortions. Our results explain the origin of the forbidden Bragg peaks recently observed in neutron diffraction experiments and reconcile the observations of strong tetragonal distortion and perfect magnetoelastic locking in Sr_{2}IrO_{4}.

Prediction of nonlinear optical properties of organic materials. General theoretical considerations

NASA Technical Reports Server (NTRS)

Cardelino, B.; Moore, C.; Zutaut, S.

1993-01-01

The prediction of nonlinear optical properties of organic materials is geared to assist materials scientists in the selection of good candidate molecules. A brief summary of the quantum mechanical methods used for estimating hyperpolarizabilities will be presented. The advantages and limitations of each technique will be discussed. Particular attention will be given to the finite-field method for calculating first and second order hyperpolarizabilities, since this method is better suited for large molecules. Corrections for dynamic fields and bulk effects will be discussed in detail, focusing on solvent effects, conformational isomerization, core effects, dispersion, and hydrogen bonding. Several results will be compared with data obtained from third-harmonic-generation (THG) and dc-induced second harmonic generation (EFISH) measurements. These comparisons will demonstrate the qualitative ability of the method to predict the relative strengths of hyperpolarizabilities of a class of compounds. The future application of molecular mechanics, as well as other techniques, in the study of bulk properties and solid state defects will be addressed. The relationship between large values for nonlinear optical properties and large conjugation lengths is well known, and is particularly important for third-order processes. For this reason, the materials with the largest observed nonresonant third-order properties are conjugated polymers. An example of this type of polymer is polydiacetylene. One of the problems in dealing with polydiacetylene is that substituents which may enhance its nonlinear properties may ultimately prevent it from polymerizing. A model which attempts to predict the likelihood of solid-state polymerization is considered, along with the implications of the assumptions that are used. Calculations of the third-order optical properties and their relationship to first-order properties and energy gaps will be discussed. The relationship between monomeric and polymeric third-order optical properties will also be considered.

Octave-spanning supercontinuum generation in in situ tapered As₂S₃ fiber pumped by a thulium-doped fiber laser.

PubMed

Rudy, Charles W; Marandi, Alireza; Vodopyanov, Konstantin L; Byer, Robert L

2013-08-01

We report a supercontinuum spanning well over an octave of measurable bandwidth from about 1 to 3.7 μm in a 2.1 mm long As₂S₃ fiber taper using the in situ tapering method. A sub-100-fs mode-locked thulium-doped fiber laser system with ~300 pJ of pulse energy was used as the pump source. Third-harmonic generation was observed and currently limits the pump pulse energy and achievable spectral bandwidth.

The Third International Symposium on Space Terahertz Technology: Symposium proceedings

NASA Technical Reports Server (NTRS)

1992-01-01

Papers from the symposium are presented that are relevant to the generation, detection, and use of the terahertz spectral region for space astronomy and remote sensing of the Earth's upper atmosphere. The program included thirteen sessions covering a wide variety of topics including solid-state oscillators, power-combining techniques, mixers, harmonic multipliers, antennas and antenna arrays, submillimeter receivers, and measurement techniques.

Centroid stabilization in alignment of FOA corner cube: designing of a matched filter

NASA Astrophysics Data System (ADS)

Awwal, Abdul; Wilhelmsen, Karl; Roberts, Randy; Leach, Richard; Miller Kamm, Victoria; Ngo, Tony; Lowe-Webb, Roger

2015-02-01

The current automation of image-based alignment of NIF high energy laser beams is providing the capability of executing multiple target shots per day. An important aspect of performing multiple shots in a day is to reduce additional time spent aligning specific beams due to perturbations in those beam images. One such alignment is beam centration through the second and third harmonic generating crystals in the final optics assembly (FOA), which employs two retro-reflecting corner cubes to represent the beam center. The FOA houses the frequency conversion crystals for third harmonic generation as the beams enters the target chamber. Beam-to-beam variations and systematic beam changes over time in the FOA corner-cube images can lead to a reduction in accuracy as well as increased convergence durations for the template based centroid detector. This work presents a systematic approach of maintaining FOA corner cube centroid templates so that stable position estimation is applied thereby leading to fast convergence of alignment control loops. In the matched filtering approach, a template is designed based on most recent images taken in the last 60 days. The results show that new filter reduces the divergence of the position estimation of FOA images.

Overcritical plasma ignition and diagnostics from oncoming interaction of two color low energy tightly focused femtosecond laser pulses inside fused silica

NASA Astrophysics Data System (ADS)

Potemkin, F. V.; Bravy, B. G.; Bezsudnova, Yu I.; Mareev, E. I.; Starostin, V. M.; Platonenko, V. T.; Gordienko, V. M.

2016-04-01

We report overcritical (3.3  ×  1021 cm-3) microplasma produced by low energy colliding IR (infrared) (1.24 μm) and visible (0.62 μm) femtosecond pulses tightly focused (NA  =  0.5) into the bulk of fused silica with on-line monitoring based on third harmonic generated by the IR beam. It was established that the absorbed energy density is the key parameter that determines the micromodification formation threshold and in our experimental conditions it is close to 4.5 kJ cm-3. Non-monotonic behavior of the third harmonic signal as a function of time delay between visible (0.62 μm) and IR (1.24 μm) femtosecond pulses demonstrates the qualitative differences about the two phenomena: one is the seed electrons generation by the visible pulse via multiphoton ionization and second is the avalanche ionization by the IR pulse. We predict that the tandem two-color excitation of wide-bandgap dielectric in comparison with single-color pulse interaction regime allows providing a much higher absorbed energy density and overcritical plasma.

High-fidelity large area nano-patterning of silicon with femtosecond light sheet

NASA Astrophysics Data System (ADS)

Sidhu, Mehra S.; Munjal, Pooja; Singh, Kamal P.

2018-01-01

We employ a femtosecond light sheet generated by a cylindrical lens to rapidly produce high-fidelity nano-structures over large area on silicon surface. The Fourier analysis of electron microscopy images of the laser-induced surface structures reveals sharp peaks indicating good homogeneity. We observed an emergence of second-order spatial periodicity on increasing the scan speed. Our reliable approach may rapidly nano-pattern curved solid surfaces and tiny objects for diverse potential applications in optical devices, structural coloring, plasmonic substrates and in high-harmonic generation.

Theoretical extension and experimental demonstration of spectral compression in second-harmonic generation by Fresnel-inspired binary phase shaping

NASA Astrophysics Data System (ADS)

Li, Baihong; Dong, Ruifang; Zhou, Conghua; Xiang, Xiao; Li, Yongfang; Zhang, Shougang

2018-05-01

Selective two-photon microscopy and high-precision nonlinear spectroscopy rely on efficient spectral compression at the desired frequency. Previously, a Fresnel-inspired binary phase shaping (FIBPS) method was theoretically proposed for spectral compression of two-photon absorption and second-harmonic generation (SHG) with a square-chirped pulse. Here, we theoretically show that the FIBPS can introduce a negative quadratic frequency phase (negative chirp) by analogy with the spatial-domain phase function of Fresnel zone plate. Thus, the previous theoretical model can be extended to the case where the pulse can be transformed limited and in any symmetrical spectral shape. As an example, we experimentally demonstrate spectral compression in SHG by FIBPS for a Gaussian transform-limited pulse and show good agreement with the theory. Given the fundamental pulse bandwidth, a narrower SHG bandwidth with relatively high intensity can be obtained by simply increasing the number of binary phases. The experimental results also verify that our method is superior to that proposed in [Phys. Rev. A 46, 2749 (1992), 10.1103/PhysRevA.46.2749]. This method will significantly facilitate the applications of selective two-photon microscopy and spectroscopy. Moreover, as it can introduce negative dispersion, hence it can also be generalized to other applications in the field of dispersion compensation.

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

NASA Astrophysics Data System (ADS)

Campbell, Kirby R.; Campagnola, Paul J.

2017-11-01

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., 10×10 microns) 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.

All-Optical Control of Linear and Nonlinear Energy Transfer via the Zeno Effect

NASA Astrophysics Data System (ADS)

Guo, Xiang; Zou, Chang-Ling; Jiang, Liang; Tang, Hong X.

2018-05-01

Microresonator-based nonlinear processes are fundamental to applications including microcomb generation, parametric frequency conversion, and harmonics generation. While nonlinear processes involving either second- (χ(2 )) or third- (χ(3 )) order nonlinearity have been extensively studied, the interaction between these two basic nonlinear processes has seldom been reported. In this paper we demonstrate a coherent interplay between second- and third- order nonlinear processes. The parametric (χ(2 ) ) coupling to a lossy ancillary mode shortens the lifetime of the target photonic mode and suppresses its density of states, preventing the photon emissions into the target photonic mode via the Zeno effect. Such an effect is then used to control the stimulated four-wave mixing process and realize a suppression ratio of 34.5.

Nonlinear electromagnetic responses of active membrane protein complexes in live cells and organelles

NASA Astrophysics Data System (ADS)

Nawarathna, Dharmakirthi

The response of biological cells to an applied oscillating electric field contains both linear and nonlinear components (eg. induced harmonics). Such noninvasive measurements can be used to study active processes taking place inside the cells. The measurement of induced harmonics is the tool used for the study described here. A highly sensitive superconducting quantum interference device (SQUID) is used to detect the response at low frequencies, which greatly reduces electrode polarization effects. At high frequencies, a four- probe method is used. At low frequencies, harmonic generation by budding yeast cells in response to a sinusoidal electric field is reported, which is seen to be minimal when the field amplitude is less than a threshold value. Surprisingly, sodium metavanadate, an inhibitor of P-type ATPases and glucose, a substrate of P-type ATPase responsible for nonlinear response in yeast, reduces the threshold field amplitude, increasing harmonic generation at low amplitudes while reducing it at large amplitudes. We have thus proposed a model that explicitly introduces a threshold field, similar to those observed in density waves, where fields above threshold drive charge transport through an energy landscape with multiple wells, and in Coulomb blockade tunnel junctions, recently exploited to define the current standard. At high frequencies, the induced harmonics exhibit pronounced features that depend on the specific organism. Budding yeast (S. cerevisiae ) cells produce numerous harmonics. When the second or third harmonic amplitude is plotted vs. applied frequency, we observe two peaks, around 3 kHz and 12 kHz, which are suppressed by the respiratory inhibitor potassium cyanide. We then measured the response to oscillatory electric fields of intact bovine heart mitochondria, a reproducible second harmonic (at ˜3-4 kHz applied frequency) was detected. Further, with coupled mouse mitochondria, an ADP sensitive peak (˜ 12-15 kHz applied frequency) was observed, possibly due to the F0 domain of ATP synthase. Finally, harmonics generated by chloroplasts, the plant organelles responsible for photosynthesis, were measured, which are similar in structure and function to mitochondria, depend dramatically on incident light, and vanish in the absence of light. Using spinach chloroplasts, light sensitive peaks were detected in the range of 0--12 kHz, again suggesting that these harmonics are indicative of electron processes in the light harvesting complexes, reaction center, and/or photosynthetic electron transport chain.

Non-critically phase-matched second harmonic generation and third order nonlinearity in organic crystal glucuronic acid γ-lactone

NASA Astrophysics Data System (ADS)

Saripalli, Ravi Kiran; Katturi, Naga Krishnakanth; Soma, Venugopal Rao; Bhat, H. L.; Elizabeth, Suja

2017-12-01

The linear, second order, and third order nonlinear optical properties of glucuronic acid γ-lactone single crystals were investigated. The optic axes and principal dielectric axes were identified through optical conoscopy and the principal refractive indices were obtained using the Brewster's angle method. Conic sections were observed which is perceived to be due to spontaneous non-collinear phase matching. The direction of collinear phase matching was determined and the deff evaluated in this direction was 0.71 pm/V. Open and closed aperture Z-scan measurements with femtosecond pulses revealed high third order nonlinearity in the form of self-defocusing, two-photon absorption, as well as saturable absorption.

Harmonic demodulation and minimum enhancement factors in field-enhanced near-field optical microscopy.

PubMed

Scarpettini, A F; Bragas, A V

2015-01-01

Field-enhanced scanning optical microscopy relies on the design and fabrication of plasmonic probes which had to provide optical and chemical contrast at the nanoscale. In order to do so, the scattering containing the near-field information recorded in a field-enhanced scanning optical microscopy experiment, has to surpass the background light, always present due to multiple interferences between the macroscopic probe and sample. In this work, we show that when the probe-sample distance is modulated with very low amplitude, the higher the harmonic demodulation is, the better the ratio between the near-field signal and the interferometric background results. The choice of working at a given n harmonic is dictated by the experiment when the signal at the n + 1 harmonic goes below the experimental noise. We demonstrate that the optical contrast comes from the nth derivative of the near-field scattering, amplified by the interferometric background. By modelling the far and near field we calculate the probe-sample approach curves, which fit very well the experimental ones. After taking a great amount of experimental data for different probes and samples, we conclude with a table of the minimum enhancement factors needed to have optical contrast with field-enhanced scanning optical microscopy. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

Photo-induced processes in collagen-hypericin system revealed by fluorescence spectroscopy and multiphoton microscopy

PubMed Central

Hovhannisyan, V.; Guo, H. W.; Hovhannisyan, A.; Ghukasyan, V.; Buryakina, T.; Chen, Y. F.; Dong, C. Y.

2014-01-01

Collagen is the main structural protein and the key determinant of mechanical and functional properties of tissues and organs. Proper balance between synthesis and degradation of collagen molecules is critical for maintaining normal physiological functions. In addition, collagen influences tumor development and drug delivery, which makes it a potential cancer therapy target. Using second harmonic generation, two-photon excited fluorescence microscopy, and spectrofluorimetry, we show that the natural pigment hypericin induces photosensitized destruction of collagen-based tissues. We demonstrate that hypericin–mediated processes in collagen fibers are irreversible and may be used for the treatment of cancer and collagen-related disorders. PMID:24877000

Photo-induced processes in collagen-hypericin system revealed by fluorescence spectroscopy and multiphoton microscopy.

PubMed

Hovhannisyan, V; Guo, H W; Hovhannisyan, A; Ghukasyan, V; Buryakina, T; Chen, Y F; Dong, C Y

2014-05-01

Collagen is the main structural protein and the key determinant of mechanical and functional properties of tissues and organs. Proper balance between synthesis and degradation of collagen molecules is critical for maintaining normal physiological functions. In addition, collagen influences tumor development and drug delivery, which makes it a potential cancer therapy target. Using second harmonic generation, two-photon excited fluorescence microscopy, and spectrofluorimetry, we show that the natural pigment hypericin induces photosensitized destruction of collagen-based tissues. We demonstrate that hypericin-mediated processes in collagen fibers are irreversible and may be used for the treatment of cancer and collagen-related disorders.

Label-free structural characterization of mitomycin C-modulated wound healing after photorefractive keratectomy by the use of multiphoton microscopy

NASA Astrophysics Data System (ADS)

Lo, Wen; Wang, Tsung-Jen; Chen, Wei-Liang; Hsueh, Chiu-Mei; Chen, Shean-Jen; Chen, Yang-Fang; Chou, Hsiu-Chu; Lin, Pi-Jung; Hu, Fung-Rong; Dong, Chen-Yuan

2010-05-01

We applied multiphoton autofluorescence (MAF) and second-harmonic generation (SHG) microscopy to monitor corneal wound healing after photorefractive keratectomy (PRK). Our results show that keratocyte activation can be observed by an increase in its MAF, while SHG imaging of corneal stroma can show the depletion of Bowman's layer after PRK and the reticular collagen deposition in the wound healing stage. Furthermore, quantification of the keratocyte activation and collagen deposition in conjunction with immunohistochemistry and histological images demonstrate the effectiveness of mitomycin C (MMC) in suppressing myofibroblast proliferation and collagen regeneration in the post-PRK wound healing process.

Experimental and simulation studies on the behavior of signal harmonics in magnetic particle imaging.

PubMed

Murase, Kenya; Konishi, Takashi; Takeuchi, Yuki; Takata, Hiroshige; Saito, Shigeyoshi

2013-07-01

Our purpose in this study was to investigate the behavior of signal harmonics in magnetic particle imaging (MPI) by experimental and simulation studies. In the experimental studies, we made an apparatus for MPI in which both a drive magnetic field (DMF) and a selection magnetic field (SMF) were generated with a Maxwell coil pair. The MPI signals from magnetic nanoparticles (MNPs) were detected with a solenoid coil. The odd- and even-numbered harmonics were calculated by Fourier transformation with or without background subtraction. The particle size of the MNPs was measured by transmission electron microscopy (TEM), dynamic light-scattering, and X-ray diffraction methods. In the simulation studies, the magnetization and particle size distribution of MNPs were assumed to obey the Langevin theory of paramagnetism and a log-normal distribution, respectively. The odd- and even-numbered harmonics were calculated by Fourier transformation under various conditions of DMF and SMF and for three different particle sizes. The behavior of the harmonics largely depended on the size of the MNPs. When we used the particle size obtained from the TEM image, the simulation results were most similar to the experimental results. The similarity between the experimental and simulation results for the even-numbered harmonics was better than that for the odd-numbered harmonics. This was considered to be due to the fact that the odd-numbered harmonics were more sensitive to background subtraction than were the even-numbered harmonics. This study will be useful for a better understanding, optimization, and development of MPI and for designing MNPs appropriate for MPI.

Optimized two- and three-colour laser pulses for the intense terahertz wave generation

NASA Astrophysics Data System (ADS)

Zhang, Lei; Wang, Guo-Li; Zhou, Xiao-Xin

2016-11-01

Based on the photocurrent model, we perform a theoretical study on the optimization of terahertz (THz) wave emission from argon gas irradiated by the two- and three-colour laser fields. To obtain stronger THz radiation for the given conditions, a genetic algorithm method is applied to search for the optimum laser parameters. For the two-colour field, our optimizations reveal two types of optimal scheme, and each one dominates the THz generation in different regions of intensity ratio for a given total laser intensity. One scheme is the combination of a fundamental laser pulse and its second harmonic, while the other is the fundamental pulse with its fourth harmonic. For each scheme, the optimal intensity ratio and phase delay are obtained. For the three-colour case, our optimization shows that the excellent waveform for the strongest THz radiation is composed of a fundamental laser pulse, and its second, third harmonics, with appropriate intensity ratio and carrier-envelope phase. Such a 3-colour field can generate strong THz radiation comparable with a 10-colour sawtooth wave [Martínez et al., Phys. Rev. Lett. 114, 183901 (2015)]. The physical mechanisms for the enhancement of THz wave emission in gases are also discussed in detail. Our results give helpful guidance for intense THz generation with tabletop femtosecond laser device in experiment.

Multimodal nonlinear imaging of arabidopsis thaliana root cell

NASA Astrophysics Data System (ADS)

Jang, Bumjoon; Lee, Sung-Ho; Woo, Sooah; Park, Jong-Hyun; Lee, Myeong Min; Park, Seung-Han

2017-07-01

Nonlinear optical microscopy has enabled the possibility to explore inside the living organisms. It utilizes ultrashort laser pulse with long wavelength (greater than 800nm). Ultrashort pulse produces high peak power to induce nonlinear optical phenomenon such as two-photon excitation fluorescence (TPEF) and harmonic generations in the medium while maintaining relatively low average energy pre area. In plant developmental biology, confocal microscopy is widely used in plant cell imaging after the development of biological fluorescence labels in mid-1990s. However, fluorescence labeling itself affects the sample and the sample deviates from intact condition especially when labelling the entire cell. In this work, we report the dynamic images of Arabidopsis thaliana root cells. This demonstrates the multimodal nonlinear optical microscopy is an effective tool for long-term plant cell imaging.

Multimodal Nonlinear Optical Microscopy

PubMed Central

Yue, Shuhua; Slipchenko, Mikhail N.; Cheng, Ji-Xin

2013-01-01

Because each nonlinear optical (NLO) imaging modality is sensitive to specific molecules or structures, multimodal NLO imaging capitalizes the potential of NLO microscopy for studies of complex biological tissues. The coupling of multiphoton fluorescence, second harmonic generation, and coherent anti-Stokes Raman scattering (CARS) has allowed investigation of a broad range of biological questions concerning lipid metabolism, cancer development, cardiovascular disease, and skin biology. Moreover, recent research shows the great potential of using CARS microscope as a platform to develop more advanced NLO modalities such as electronic-resonance-enhanced four-wave mixing, stimulated Raman scattering, and pump-probe microscopy. This article reviews the various approaches developed for realization of multimodal NLO imaging as well as developments of new NLO modalities on a CARS microscope. Applications to various aspects of biological and biomedical research are discussed. PMID:24353747

Application of a reflective microscope objective for multiphoton microscopy.

PubMed

Kabir, Mohammad M; Choubal, Aakash M; Toussaint, Kimani C

2018-04-20

Reflective objectives (ROs) mitigate chromatic aberration across a broad wavelength range. Yet, a systematic performance characterisation of ROs has not been done. In this paper, we compare the performance of a 0.5 numerical-aperture (NA) reflective objective (RO) with a 0.55 NA standard glass objective (SO), using two-photon fluorescence (TPF) and second-harmonic generation (SHG). For experiments spanning ∼1 octave in the visible and NIR wavelengths, the SO leads to defocusing errors of 25-40% for TPF images of subdiffraction fluorescent beads and 10-12% for SHG images of collagen fibres. The corresponding error for the RO is ∼4% for both imaging modalities. This work emphasises the potential utility of ROs for multimodal multiphoton microscopy applications. © 2018 The Authors Journal of Microscopy © 2018 Royal Microscopical Society.

Double-clad photonic crystal fiber coupler for compact nonlinear optical microscopy imaging.

PubMed

Fu, Ling; Gu, Min

2006-05-15

A 1 x 2 double-clad photonic crystal fiber coupler is fabricated by the fused tapered method, showing a low excess loss of 1.1 dB and a splitting ratio of 97/3 over the entire visible and near-infrared wavelength range. In addition to the property of splitting the laser power, the double-clad feature of the coupler facilitates the separation of a near-infrared single-mode beam from a visible multimode beam, which is ideal for nonlinear optical microscopy imaging. In conjunction with a gradient-index lens, this coupler is used to construct a miniaturized microscope based on two-photon fluorescence and second-harmonic generation. Three-dimensional nonlinear optical images demonstrate potential applications of the coupler to compact all-fiber and nonlinear optical microscopy and endoscopy.

An optical parametric chirped-pulse amplifier for seeding high repetition rate free-electron lasers

DOE PAGES

Höppner, H.; Hage, A.; Tanikawa, T.; ...

2015-05-15

High repetition rate free-electron lasers (FEL), producing highly intense extreme ultraviolet and x-ray pulses, require new high power tunable femtosecond lasers for FEL seeding and FEL pump-probe experiments. A tunable, 112 W (burst mode) optical parametric chirped-pulse amplifier (OPCPA) is demonstrated with center frequencies ranging from 720–900 nm, pulse energies up to 1.12 mJ and a pulse duration of 30 fs at a repetition rate of 100 kHz. Since the power scalability of this OPCPA is limited by the OPCPA-pump amplifier, we also demonstrate a 6.7–13.7 kW (burst mode) thin-disk OPCPA-pump amplifier, increasing the possible OPCPA output power to manymore » hundreds of watts. Furthermore, third and fourth harmonic generation experiments are performed and the results are used to simulate a seeded FEL with high-gain harmonic generation.« less

Two-photon Microscopy and Polarimetry for Assessment of Myocardial Tissue Organization

NASA Astrophysics Data System (ADS)

Archambault-Wallenburg, Marika

Optical methods can provide useful tissue characterization tools. For this project, two-photon microscopy and polarized light examinations (polarimetry) were used to assess the organizational state of myocardium in healthy, infarcted, and stem-cell regenerated states. Two-photon microscopy visualizes collagen through second-harmonic generation and myocytes through two-photon excitation autofluorescence, providing information on the composition and structure/organization of the tissue. Polarimetry measurements yield a value of linear retardance that can serve as an indicator of tissue anisotropy, and with a dual-projection method, information about the anisotropy axis orientation can also be extracted. Two-photon microscopy results reveal that stem-cell treated tissue retains more myocytes and structure than infarcted myocardium, while polarimetry findings suggest that the injury caused by temporary ligation of a coronary artery is less severe and more diffuse that than caused by a permanent ligation. Both these methods show potential for tissue characterization.

Extracellular localization of the diterpene sclareol in clary sage (Salvia sclarea L., Lamiaceae).

PubMed

Caissard, Jean-Claude; Olivier, Thomas; Delbecque, Claire; Palle, Sabine; Garry, Pierre-Philippe; Audran, Arthur; Valot, Nadine; Moja, Sandrine; Nicolé, Florence; Magnard, Jean-Louis; Legrand, Sylvain; Baudino, Sylvie; Jullien, Frédéric

2012-01-01

Sclareol is a high-value natural product obtained by solid/liquid extraction of clary sage (Salvia sclarea L.) inflorescences. Because processes of excretion and accumulation of this labdane diterpene are unknown, the aim of this work was to gain knowledge on its sites of accumulation in planta. Samples were collected in natura or during different steps of the industrial process of extraction (steam distillation and solid/liquid extraction). Samples were then analysed with a combination of complementary analytical techniques (gas chromatography coupled to a mass spectrometer, polarized light microscopy, environmental scanning electron microscopy, two-photon fluorescence microscopy, second harmonic generation microscopy). According to the literature, it is hypothesized that sclareol is localized in oil pockets of secretory trichomes. This study demonstrates that this is not the case and that sclareol accumulates in a crystalline epicuticular form, mostly on calyces.

Extracellular Localization of the Diterpene Sclareol in Clary Sage (Salvia sclarea L., Lamiaceae)

PubMed Central

Caissard, Jean-Claude; Olivier, Thomas; Delbecque, Claire; Palle, Sabine; Garry, Pierre-Philippe; Audran, Arthur; Valot, Nadine; Moja, Sandrine; Nicolé, Florence; Magnard, Jean-Louis; Legrand, Sylvain; Baudino, Sylvie; Jullien, Frédéric

2012-01-01

Sclareol is a high-value natural product obtained by solid/liquid extraction of clary sage (Salvia sclarea L.) inflorescences. Because processes of excretion and accumulation of this labdane diterpene are unknown, the aim of this work was to gain knowledge on its sites of accumulation in planta. Samples were collected in natura or during different steps of the industrial process of extraction (steam distillation and solid/liquid extraction). Samples were then analysed with a combination of complementary analytical techniques (gas chromatography coupled to a mass spectrometer, polarized light microscopy, environmental scanning electron microscopy, two-photon fluorescence microscopy, second harmonic generation microscopy). According to the literature, it is hypothesized that sclareol is localized in oil pockets of secretory trichomes. This study demonstrates that this is not the case and that sclareol accumulates in a crystalline epicuticular form, mostly on calyces. PMID:23133579

Development of a Fiber Laser with Independently Adjustable Properties for Optical Resolution Photoacoustic Microscopy.

PubMed

Aytac-Kipergil, Esra; Demirkiran, Aytac; Uluc, Nasire; Yavas, Seydi; Kayikcioglu, Tunc; Salman, Sarper; Karamuk, Sohret Gorkem; Ilday, Fatih Omer; Unlu, Mehmet Burcin

2016-12-08

Photoacoustic imaging is based on the detection of generated acoustic waves through thermal expansion of tissue illuminated by short laser pulses. Fiber lasers as an excitation source for photoacoustic imaging have recently been preferred for their high repetition frequencies. Here, we report a unique fiber laser developed specifically for multiwavelength photoacoustic microscopy system. The laser is custom-made for maximum flexibility in adjustment of its parameters; pulse duration (5-10 ns), pulse energy (up to 10 μJ) and repetition frequency (up to 1 MHz) independently from each other and covers a broad spectral region from 450 to 1100 nm and also can emit wavelengths of 532, 355, and 266 nm. The laser system consists of a master oscillator power amplifier, seeding two stages; supercontinuum and harmonic generation units. The laser is outstanding since the oscillator, amplifier and supercontinuum generation parts are all-fiber integrated with custom-developed electronics and software. To demonstrate the feasibility of the system, the images of several elements of standardized resolution test chart are acquired at multiple wavelengths. The lateral resolution of optical resolution photoacoustic microscopy system is determined as 2.68 μm. The developed system may pave the way for spectroscopic photoacoustic microscopy applications via widely tunable fiber laser technologies.

Development of a Fiber Laser with Independently Adjustable Properties for Optical Resolution Photoacoustic Microscopy

PubMed Central

Aytac-Kipergil, Esra; Demirkiran, Aytac; Uluc, Nasire; Yavas, Seydi; Kayikcioglu, Tunc; Salman, Sarper; Karamuk, Sohret Gorkem; Ilday, Fatih Omer; Unlu, Mehmet Burcin

2016-01-01

Photoacoustic imaging is based on the detection of generated acoustic waves through thermal expansion of tissue illuminated by short laser pulses. Fiber lasers as an excitation source for photoacoustic imaging have recently been preferred for their high repetition frequencies. Here, we report a unique fiber laser developed specifically for multiwavelength photoacoustic microscopy system. The laser is custom-made for maximum flexibility in adjustment of its parameters; pulse duration (5–10 ns), pulse energy (up to 10 μJ) and repetition frequency (up to 1 MHz) independently from each other and covers a broad spectral region from 450 to 1100 nm and also can emit wavelengths of 532, 355, and 266 nm. The laser system consists of a master oscillator power amplifier, seeding two stages; supercontinuum and harmonic generation units. The laser is outstanding since the oscillator, amplifier and supercontinuum generation parts are all-fiber integrated with custom-developed electronics and software. To demonstrate the feasibility of the system, the images of several elements of standardized resolution test chart are acquired at multiple wavelengths. The lateral resolution of optical resolution photoacoustic microscopy system is determined as 2.68 μm. The developed system may pave the way for spectroscopic photoacoustic microscopy applications via widely tunable fiber laser technologies. PMID:27929049

Characterisation of the dynamic behaviour of lipid droplets in the early mouse embryo using adaptive harmonic generation microscopy.

PubMed

Watanabe, Tomoko; Thayil, Anisha; Jesacher, Alexander; Grieve, Kate; Debarre, Delphine; Wilson, Tony; Booth, Martin; Srinivas, Shankar

2010-06-03

Lipid droplets (LD) are organelles with an important role in normal metabolism and disease. The lipid content of embryos has a major impact on viability and development. LD in Drosophila embryos and cultured cell lines have been shown to move and fuse in a microtubule dependent manner. Due to limitations in current imaging technology, little is known about the behaviour of LD in the mammalian embryo. Harmonic generation microscopy (HGM) allows one to image LD without the use of exogenous labels. Adaptive optics can be used to correct aberrations that would otherwise degrade the quality and information content of images. We have built a harmonic generation microscope with adaptive optics to characterise early mouse embryogenesis. At fertilization, LD are small and uniformly distributed, but in the implanting blastocyst, LD are larger and enriched in the invading giant cells of the trophectoderm. Time-lapse studies reveal that LD move continuously and collide but do not fuse, instead forming aggregates that subsequently behave as single units. Using specific inhibitors, we show that the velocity and dynamic behaviour of LD is dependent not only on microtubules as in other systems, but also on microfilaments. We explore the limits within which HGM can be used to study living embryos without compromising viability and make the counterintuitive finding that 16 J of energy delivered continuously over a period of minutes can be less deleterious than an order of magnitude lower energy delivered dis-continuously over a period of hours. LD in pre-implantation mouse embryos show a previously unappreciated complexity of behaviour that is dependent not only on microtubules, but also microfilaments. Unlike LD in other systems, LD in the mouse embryo do not fuse but form aggregates. This study establishes HGM with adaptive optics as a powerful tool for the study of LD biology and provides insights into the photo-toxic effects of imaging embryos.

Ultrasound contrast agent imaging: Real-time imaging of the superharmonics

NASA Astrophysics Data System (ADS)

Peruzzini, D.; Viti, J.; Tortoli, P.; Verweij, M. D.; de Jong, N.; Vos, H. J.

2015-10-01

Currently, in medical ultrasound contrast agent (UCA) imaging the second harmonic scattering of the microbubbles is regularly used. This scattering is in competition with the signal that is caused by nonlinear wave propagation in tissue. It was reported that UCA imaging based on the third or higher harmonics, i.e. "superharmonic" imaging, shows better contrast. However, the superharmonic scattering has a lower signal level compared to e.g. second harmonic signals. This study investigates the contrast-to-tissue ratio (CTR) and signal to noise ratio (SNR) of superharmonic UCA scattering in a tissue/vessel mimicking phantom using a real-time clinical scanner. Numerical simulations were performed to estimate the level of harmonics generated by the microbubbles. Data were acquired with a custom built dual-frequency cardiac phased array probe. Fundamental real-time images were produced while beam formed radiofrequency (RF) data was stored for further offline processing. The phantom consisted of a cavity filled with UCA surrounded by tissue mimicking material. The acoustic pressure in the cavity of the phantom was 110 kPa (MI = 0.11) ensuring non-destructivity of UCA. After processing of the acquired data from the phantom, the UCA-filled cavity could be clearly observed in the images, while tissue signals were suppressed at or below the noise floor. The measured CTR values were 36 dB, >38 dB, and >32 dB, for the second, third, and fourth harmonic respectively, which were in agreement with those reported earlier for preliminary contrast superharmonic imaging. The single frame SNR values (in which `signal' denotes the signal level from the UCA area) were 23 dB, 18 dB, and 11 dB, respectively. This indicates that noise, and not the tissue signal, is the limiting factor for the UCA detection when using the superharmonics in nondestructive mode.

Long-term in vivo study of vertebrate embryonic development using noninvasive harmonics optical microscopy

NASA Astrophysics Data System (ADS)

Chen, Szu-Yu; Hsieh, C.-S.; Chu, S.-W.; Lin, Cheng-Yung; Ko, C.-Y.; Chen, Y.-C.; Tsai, Huai-Jen; Hu, C.-H.; Sun, Chi-Kuang

2005-03-01

Harmonics optical microscopy (HOM) provides a truly "noninvasive" tool for in vivo and long-term study of vertebrate embryonic development. Based on the nonlinear natures, it provides sub-micrometer 3D spatial resolution and high 3D optical-sectioning power (~1μm axial resolution) without using invasive and toxic fluorophores. Since only virtual-level-transition is involved, HOM is known to leave no energy deposition and no photodamages. Combined with second harmonic generation, which is sensitive to specific structure such as nerve and muscle fibers, HOM can be used to do functional studies of early developmental dynamics of many vertebrate physiological systems. Recently, zebrafish has become a standard model for many biological and medical studies of vertebrates, due to the similarity between embryonic development of zebrafish and human being. Zebrafish embryos now have been used to study many vertebrate physiological systems. We have demonstrated an in vivo HOM study of developmental dynamics of several embryonic physiological systems in live zebrafish embryos, with focuses on the developments of brains, eyes, ears, and hearts. Based on a femtosecond Cr:forsterite laser, which provides the deepest penetration (~1.5mm) and least photodamage in the zebrafish embryo, complete developing processes of different physiological systems within a period of time longer than 20 hours can be non-invasively observed inside the same embryo.

Multimodal second harmonic generation and two photon fluorescence imaging of microdomain calcium contraction coupling in single cardiomyocytes

NASA Astrophysics Data System (ADS)

Chan, James; Awasthi, Samir; Izu, Leighton; Mao, Ziliang; Jian, Zhong; Landas, Trevor; Lerner, Aaron; Shimkunas, Rafael; Woldeyesus, Rahwa; Bossuyt, Julie; Wood, Brittani; Chen, Yi-Je; Matthews, Dennis; Lieu, Deborah; Chiamvimonvat, Nipavan; Lam, Kit; Chen-Izu, Ye

2016-11-01

The objective of this study was to develop a method for simultaneously measuring the calcium and contraction dynamics of single, live cardiomyocytes at high spatial resolutions. Such measurements are important to investigate local calcium release and the mechanical response at the sarcomere level (i.e. the basic unit of contraction), which have important implications in cardiac dysfunction and arrhythmias in conditions such as hypertension, atrial fibrillation, and myocardial infarction. Here, we describe a multimodal second harmonic generation (SHG) and two photon fluorescence (2PF) microscopy technique that is used to simultaneously measure subsarcomere calcium and contraction events at high spatial and temporal resolutions. The method takes advantage of the label-free nature of SHG for imaging the sarcomeres and the high spatial colocalization of the SHG signal and the fluorescence signal excited from calcium indicators. This microscope was used to measure calcium sparks and waves and associated contractions in subcellular microdomains, leading to the generation of subcellular strain. We anticipate this new imaging tool will play an important role in studying mechanical stress-induced heart disease.

Laser scanning saturated structured illumination microscopy based on phase modulation

NASA Astrophysics Data System (ADS)

Huang, Yujia; Zhu, Dazhao; Jin, Luhong; Kuang, Cuifang; Xu, Yingke; Liu, Xu

2017-08-01

Wide-field saturated structured illumination microscopy has not been widely used due to the requirement of high laser power. We propose a novel method called laser scanning saturated structured illumination microscopy (LS-SSIM), which introduces high order of harmonics frequency and greatly reduces the required laser power for SSIM imaging. To accomplish that, an excitation PSF with two peaks is generated and scanned along different directions on the sample. Raw images are recorded cumulatively by a CCD detector and then reconstructed to form a high-resolution image with extended optical transfer function (OTF). Our theoretical analysis and simulation results show that LS-SSIM method reaches a resolution of 0.16 λ, equivalent to 2.7-fold resolution than conventional wide-field microscopy. In addition, LS-SSIM greatly improves the optical sectioning capability of conventional wide-field illumination system by diminishing our-of-focus light. Furthermore, this modality has the advantage of implementation in multi-photon microscopy with point scanning excitation to image samples in greater depths.

Fibrillar collagen scoring by second harmonic microscopy: a new tool in the assessment of liver fibrosis.

PubMed

Gailhouste, Luc; Le Grand, Yann; Odin, Christophe; Guyader, Dominique; Turlin, Bruno; Ezan, Frédéric; Désille, Yoann; Guilbert, Thomas; Bessard, Anne; Frémin, Christophe; Theret, Nathalie; Baffet, Georges

2010-03-01

Imaging of supramolecular structures by multiphoton microscopy offers significant advantages for studying specific fibrillar compounds in biological tissues. In this study, we aimed to demonstrate the relevance of Second Harmonic Generation (SHG) for assessing and quantifying, without staining, fibrillar collagen in liver fibrosis. We first showed the relationship between SHG signal and collagen forms over-produced and accumulated during fibrosis progression. Taking this property into consideration, we developed an innovative method to precisely quantify the fibrosis area in histological slices by scoring of fibrillar collagen deposits (Fibrosis-SHG index). The scoring method was routinely applied to 119 biopsies from patients with chronic liver disease allowing a fast and accurate measurement of fibrosis correlated with the Fibrosis-Metavir score (rho=0.75, p<0.0001). The technique allowed discriminating patients with advanced (moderate to severe) fibrosis (AUROC=0.88, p<0.0001) and cirrhosis (AUROC=0.89, p<0.0001). Taking advantage of its continuous gradation, the Fibrosis-SHG index also allowed the discrimination of several levels of fibrosis within the same F-Metavir stage. The SHG process presented several advantages such as a high reliability and sensitivity that lead to a standardized evaluation of hepatic fibrosis in liver biopsies without staining and pathological examination. Second harmonic microscopy emerges as an original and powerful tool in the assessment of liver fibrosis and offers new possibilities for the evaluation of experimental protocols. We expect that this technology could easily be applicable in the study of other fibro-proliferative pathologies. Copyright (c) 2010 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Efficient optical nonlinear Langmuir-Blodgett films: roles of matrix molecules

NASA Astrophysics Data System (ADS)

Ma, Shihong; Lu, Xingze; Liu, Liying; Han, Kui; Wang, Wencheng; Zhang, Zhi-Ming

1996-10-01

A novel bifat-chain amphiphilic molecule nitrogencrown (NC) was adopted as an inert material for fabrication of optical nonlinear Langmuir-Blodgett (LB) multilayers. Structural improvement in the Z-type mixed fullerene derivative (C60-Be)/NC LB multilayers samples was realized by insertion of the C60-Be molecules between two hydrophobic chains of the NC molecules. The relatively large third-order susceptibility (chi) (3)xxxx(- 3(omega) ;(omega) ,(omega) ,(omega) ) equals 2.9 multiplied by 10-19 M2V-2 (or 2.1 multiplied by 10-11 esu) was deduced by measuring third harmonic generation (THG) from the C60-Be samples. The second harmonic generation (SHG) intensity increased quadratically with the bilayer number (up to 116 bilayers) in Y-type hemicyanine (HEM)/NC interleaving LB multilayers due to improvement of the structural properties by insertion of the long hydrophobic tail of HEM molecules between two chains of NC molecules. The second-order susceptibility (chi) (2)zxx(-2(omega) ;(omega) ,(omega) ) equals 18 pM V-1 (or 4.35 multiplied by 10-8 esu) was obtained by measuring SHG from the HEM samples. The NC molecule has attractive features as a matrix material in fabrications of LB multilayers made from optically nonlinear materials with hydrophobic long tails or ball-like molecules.

A novel intravital multi-harmonic generation microscope for early diagnosis of oral cancer

NASA Astrophysics Data System (ADS)

Cheng, Yu-Hsiang; Lin, Chih-Feng; Shih, Ting-Fang; Sun, Chi-Kuang

2013-03-01

Oral cancer is one of the most frequently diagnosed human cancers and leading causes of cancer death all over the world, but the prognosis and overall survival rate are still poor because of delay in diagnosis and lack of early intervention. The failure of early diagnosis is due to insufficiency of proper diagnostic and screening tools and most patients are reluctant to undergo biopsy. Optical virtual biopsy techniques, for imaging cells and tissues at microscopic details capable of differentiating benign from malignant lesions non-invasively, are thus highly desirable. A novel multi-harmonic generation microscope, excited by a 1260 nm Cr:forsterite laser, with second and third harmonic signals demonstrating collagen fiber distribution and cell morphology in a sub-micron resolution, was developed for clinical use. To achieve invivo observation inside the human oral cavity, a small objective probe with a suction capability was carefully designed for patients' comfort and stability. By remotely changing its focus point, the same objective can image the mucosa surface with a low magnification, illuminated by side light-emitting diodes, with a charge-coupled device (CCD) for site location selection before the harmonic generation biopsy was applied. Furthermore, the slow galvanometer mirror and the fast resonant mirror provide a 30 fps frame rate for high-speed real-time observation and the z-motor of this system is triggered at the same rate to provide fast 3D scanning, again ensuring patients' comfort. Focusing on the special cytological and morphological changes of the oral epithelial cells, our preliminary result disclosed excellent consistency with traditional histopathology studies.

Distinguishing human normal or cancerous esophagus tissue ex vivo using multiphoton microscopy

NASA Astrophysics Data System (ADS)

Liu, N. R.; Chen, G. N.; Wu, S. S.; Chen, R.

2014-02-01

Application of multiphoton microscopy (MPM) to clinical cancer research has greatly developed over the last few years. In this paper, we mainly focus on two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG) for investigating esophageal cancer. We chiefly discuss the SHG/TPEF image and spectral characteristics of normal and cancerous esophagus submucosa with the combined multi-channel imaging mode and Lambda mode of a multiphoton microscope (LSM 510 META). Great differences can be detected, such as collagen content and morphology, glandular-shaped cancer cells, TPEF/SHG intensity ratio, and so on, which demonstrate that the multiphoton imaging technique has the potential ability for minimally-invasive early cancer diagnosis.

Insights into esophagus tissue architecture using two-photon confocal microscopy

NASA Astrophysics Data System (ADS)

Liu, Nenrong; Wang, Yue; Feng, Shangyuan; Chen, Rong

2013-08-01

In this paper, microstructures of human esophageal mucosa were evaluated using the two-photon laser scanning confocal microscopy (TPLSCM), based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). The distribution of epithelial cells, muscle fibers of muscularis mucosae has been distinctly obtained. Furthermore, esophageal submucosa characteristics with cancer cells invading into were detected. The variation of collagen, elastin and cancer cells is very relevant to the pathology in esophagus, especially early esophageal cancer. Our experimental results indicate that the MPM technique has the much more advantages for label-free imaging, and has the potential application in vivo in the clinical diagnosis and monitoring of early esophageal cancer.

Second harmonic generation polarization properties of myofilaments

NASA Astrophysics Data System (ADS)

Samim, Masood; Prent, Nicole; Dicenzo, Daniel; Stewart, Bryan; Barzda, Virginijus

2014-05-01

Second harmonic generation (SHG) polarization microscopy was used to investigate the organization of myosin nanomotors in myofilaments of muscle cells. The distribution of the second-order nonlinear susceptibility component ratio χzzz(2)/χzxx(2) along anisotropic bands of sarcomeres revealed differences between the headless and head-containing regions of myofilaments. The polarization-in polarization-out SHG measurements of headless myosin mutants of indirect flight muscle in Drosophila melanogaster confirmed a lower susceptibility component ratio compared to the head-containing myocytes with wild-type myosins. The increase in the ratio is assigned to the change in the deflection angle of the myosin S2 domain and possible contribution of myosin heads. The nonlinear susceptibility component ratio is a sensitive indicator of the myosin structure, and therefore, it can be used for conformational studies of myosin nanomotors. The measured ratio values can also be used as the reference for ab initio calculations of nonlinear optical properties of different parts of myosins.

Imaging rat esophagus using combination of reflectance confocal and multiphoton microscopy

NASA Astrophysics Data System (ADS)

Zhuo, S. M.; Chen, J. X.; Jiang, X. S.; Lu, K. C.; Xie, S. S.

2008-08-01

We combine reflectance confocal microscopy (RCM) with multiphoton microscopy (MPM) to image rat esophagus. The two imaging modalities allow detection of layered-resolved complementary information from esophagus. In the keratinizing layer, the keratinocytes boundaries can be characterized by RCM, while the keratinocytes cytoplasm (keratin) can be further imaged by multiphoton autofluorescence signal. In the epithelium, the epithelial cellular boundaries and nucleus can be detected by RCM, and MPM can be used for imaging epithelial cell cytoplasm and monitoring metabolic state of epithelium. In the stroma, multiphoton autofluorescence signal is used to image elastin and second harmonic generation signal is utilized to detect collagen, while RCM is used to determine the optical property of stroma. Overall, these results suggest that the combination of RCM and MPM has potential to provide more important and comprehensive information for early diagnosis of esophageal cancer.

Generation Mechanism of Nonlinear Rayleigh Surface Waves for Randomly Distributed Surface Micro-Cracks.

PubMed

Ding, Xiangyan; Li, Feilong; Zhao, Youxuan; Xu, Yongmei; Hu, Ning; Cao, Peng; Deng, Mingxi

2018-04-23

This paper investigates the propagation of Rayleigh surface waves in structures with randomly distributed surface micro-cracks using numerical simulations. The results revealed a significant ultrasonic nonlinear effect caused by the surface micro-cracks, which is mainly represented by a second harmonic with even more distinct third/quadruple harmonics. Based on statistical analysis from the numerous results of random micro-crack models, it is clearly found that the acoustic nonlinear parameter increases linearly with micro-crack density, the proportion of surface cracks, the size of micro-crack zone, and the excitation frequency. This study theoretically reveals that nonlinear Rayleigh surface waves are feasible for use in quantitatively identifying the physical characteristics of surface micro-cracks in structures.

Ultrafast frequency-selective optical switching based on thin self-assembled organic chromophoric films with a large second-order nonlinear response

NASA Astrophysics Data System (ADS)

Wang, Gang; Zhu, Peiwang; Marks, Tobin J.; Ketterson, J. B.

2002-09-01

Thin films consisting of self-assembled chromophoric superlattices exhibit very large second-order nonlinear responses [chi](2). Using such films, a "static" diffraction grating is created by the interference of two coherent infrared beams from a pulsed yttritium-aluminum-garnet laser. This grating is used to switch the second-harmonic and third-harmonic "signal" beams (generated from the fundamental "pump" beam or mixed within the chromophoric superlattice) into different channels (directions). Ultrafast switching response as a function of the time overlap of the pumping beams is demonstrated. It is suggested that such devices can be used to spatially and temporally separate signal trains consisting of pulses having different frequencies and arrival times.

Generation Mechanism of Nonlinear Rayleigh Surface Waves for Randomly Distributed Surface Micro-Cracks

PubMed Central

Ding, Xiangyan; Li, Feilong; Xu, Yongmei; Cao, Peng; Deng, Mingxi

2018-01-01

This paper investigates the propagation of Rayleigh surface waves in structures with randomly distributed surface micro-cracks using numerical simulations. The results revealed a significant ultrasonic nonlinear effect caused by the surface micro-cracks, which is mainly represented by a second harmonic with even more distinct third/quadruple harmonics. Based on statistical analysis from the numerous results of random micro-crack models, it is clearly found that the acoustic nonlinear parameter increases linearly with micro-crack density, the proportion of surface cracks, the size of micro-crack zone, and the excitation frequency. This study theoretically reveals that nonlinear Rayleigh surface waves are feasible for use in quantitatively identifying the physical characteristics of surface micro-cracks in structures. PMID:29690580

Investigation on fibrous collagen modifications during corneal laser welding by second harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Matteini, Paolo; Ratto, Fulvio; Rossi, Francesca; Cicchi, Riccardo; Stringari, Chiara; Kapsokalyvas, Dimitrios; Pavone, Francesco S.; Pini, Roberto

2009-02-01

The structural modifications in the collagen lattice of corneal stroma induced by near-infrared laser welding were investigated with second-harmonic generation (SHG) imaging. The corneal laser welding procedure is performed by staining the wound edges with a saturated water solution of Indocyanine Green (ICG) followed by irradiation with a 810 nm diode laser operated in continuous (CWLW: continuous wave laser welding) or pulsed (PLW: pulsed laser welding) mode. Both these procedures can provide closure of corneal wounds by inducing different structural modifications in the extracellular matrix. SHG imaging of native corneal stroma revealed collagen bundles composed of many regularly aligned collagen fibrils. After CWLW the regular lamellar arrangement was lost; collagen bundles appeared densely packed with an increasing disordered arrangement toward the welded cut. The weld was characterized by a loss of details; nevertheless, the observation of the second harmonic signal at this site indicated the lack of collagen denaturation. By contrast, PLW mode produced welding spots at the interface between donor and recipient corneal layers, which were characterized by a severe loss of the SHG signal, suggesting the occurrence of a complete collagen denaturation. SHG imaging appeared to be a powerful tool for visualizing the supramolecular morphological modifications in the collagen matrix after laser welding.

Relaxor ferroelectricity, ferromagnetic and optical second harmonic properties in lanthanum lithium niobate (La0.05Li0.85NbO3) nanoparticles

NASA Astrophysics Data System (ADS)

Díaz-Moreno, Carlos A.; Ding, Yu; Li, Chunqiang; Portelles, Jorge; Heiras, J.; Hurtado-Macias, A.; Farias, J. R.; González-Hernández, J.; Yacamán, M. J.; López, Jorge

2017-07-01

Relaxor ferroelectricity, ferromagnetism and Second Harmonic Generation properties were founded and studied as a function of a reduction heat treatment at 650 °C in a Ar-5%H2 atmosphere in stoichiometric La0.05Li0.85NbO3 nanoparticles of 40 nm. A diffuse dielectric anomaly related with relaxor behavior from 25 °C to 800 °C in a frequency range from 100 Hz to 1 MHz was founded. It also shows ferromagnetic anhysterestic type and ferroelectric hysteresis loops at room temperature with a magnetic spin remnant of 2.5 × 10-3 emu/g and polarization saturation of 0.235 μC/cm2, remnant polarization of 0.141 μC/cm2, coercive field of 1.35 kV/cm, respectively. It shows very good second harmonic generation signal at 450 nm and 500 nm. High Resolution Transmission Electron Microscopy, X-ray Photoelectron Spectroscopy and Raman spectroscopy, indicate an ABO3 perovskite structure, new electronic binding energy structure for La (5s, 4d), Li (1s), Nb (4s, 3d, 4p) and oxygen (1s, 2s) and new vibrations modes on octahedron NbO6 related to multiferroic single phase nanoparticles, respectively.

Determination of nonlinear resistance voltage-current relationships by measuring harmonics

NASA Technical Reports Server (NTRS)

Stafford, J. M.

1971-01-01

Test configuration measures harmonic signal amplitudes generated in nonlinear resistance. Vacuum-type voltmeter measures low frequency sinusoidal input signal amplitude and wave-analyzer measures amplitude of harmonic signals generated in junction. Input signal harmonics amplitude must not exceed that of harmonics generated in nonlinear resistance.

Effect of laser polarization and pulse energy on therapeutic, femtosecond laser-induced second harmonic generation in corneal tissue (Conference Presentation)

NASA Astrophysics Data System (ADS)

Calhoun, William R.; Ilev, Ilko K.

2016-03-01

Some of the most commonly performed surgical operations in the world, including laser-assisted in-situ keratomileusis (LASIK), lens replacement (e.g. cataract surgery), and keratoplasty (cornea transplant), now employ therapeutic infrared femtosecond lasers (FSLs) for their extreme precision, low energy delivered into tissue and advanced ablation characteristics. Although the widely exploited applications of FSLs in medical therapeutics offer significant benefits, FSLs must generate very high intensities in order to achieve optical breakdown, the predominant tissue ablative mechanism, which can also stimulate nonlinear optical effects such as harmonic generation, an effect that generates coherent visible and UV light in the case of second- (SHG) and third-harmonic generation (THG), respectively. In order to improve the understanding of HG in corneal tissue, the effect of FSL polarization and pulse energy were investigated. FSL stimulated SHG intensity in corneal tissue was measured as the laser polarization was rotated 360 degrees. Further, the pulse energy at the SHG wavelength were measured for single FSL pulses as the pulse energy at the fundamental wavelength was varied through a range of clinically relevant values. The results of this study revealed SHG intensity oscillated with laser polarization, having a variation greater than 20%. This relationship seems to due to the intrinsic anisotropy of collagen fibril hyperpolarizability, not related to tissue birefringence. SHG pulse energy measurements showed an increase in SHG pulse energy with increasing FSL pulse energy, however conversion efficiency decreased. This may be related to the dynamic relationship between optical breakdown leading to tissue destruction and HG evolution.

First-harmonic nonlinearities can predict unseen third-harmonics in medium-amplitude oscillatory shear (MAOS)

NASA Astrophysics Data System (ADS)

Carey-De La Torre, Olivia; Ewoldt, Randy H.

2018-02-01

We use first-harmonic MAOS nonlinearities from G 1' and G 1″ to test a predictive structure-rheology model for a transient polymer network. Using experiments with PVA-Borax (polyvinyl alcohol cross-linked by sodium tetraborate (borax)) at 11 different compositions, the model is calibrated to first-harmonic MAOS data on a torque-controlled rheometer at a fixed frequency, and used to predict third-harmonic MAOS on a displacement controlled rheometer at a different frequency three times larger. The prediction matches experiments for decomposed MAOS measures [ e 3] and [ v 3] with median disagreement of 13% and 25%, respectively, across all 11 compositions tested. This supports the validity of this model, and demonstrates the value of using all four MAOS signatures to understand and test structure-rheology relations for complex fluids.

Biological Effects of Laser Radiation. Volume IV. Optical Second Harmonic Generation in Biological Tissues.

DTIC Science & Technology

1978-10-17

characteristics for optical second- harmonic generation. The collage component of conective tissue may be the principal site for the observed harmonic...Generation in Tissue ; Second Harmonic Generation in Collage; Glutathione, 5MB; Mechanisms; Conversion Efficiency; Significance of order UL AIM UY#m~wmev...sclera, and skin on 694 im. Q-switched ruby laser irradiation. A possible source of this second-harmonic generation was tissue collagen; because of

High Harmonic Radiation Generation and Attosecond pulse generation from Intense Laser-Solid Interactions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Thomas, Alexander Roy; Krushelnick, Karl

2016-09-08

We have studied ion motion effects in high harmonic generation, including shifts to the harmonics which result in degradation of the attosecond pulse train, and how to mitigate them. We have examined the scaling with intensity of harmonic emission. We have also switched the geometry of the interaction to measure, for the first time, harmonics from a normal incidence interaction. This was performed by using a special parabolic reflector with an on axis hole and is to allow measurements of the attosecond pulses using standard techniques. Here is a summary of the findings: First high harmonic generation in laser-solid interactionsmore » at 10 21 Wcm -2, demonstration of harmonic focusing, study of ion motion effects in high harmonic generation in laser-solid interactions, and demonstration of harmonic amplification.« less

DIRECT SIMULATION OF A-C MACHINERY.

DTIC Science & Technology

show the application of the simulation to both induction and synchronous machines. The fundamental space harmonic only, the fundamental and third ... space harmonic only, or all the space harmonics are considered. The report concludes that: (1) Successful direct simulation of the 2-phase induction

Investigation of superharmonic sound propagation and imaging in biological tissues in vitro.

PubMed

Ma, Qingyu; Zhang, Dong; Gong, Xiufen; Ma, Yong

2006-04-01

This article presents both theoretical and experimental studies on the superharmonic generation and its imaging in biological tissues. A superharmonic component is defined as a summation of the third-, fourth-, and fifth-order harmonics. A superharmonic signal is produced using an 8-mm-diam, 2.5-MHz planar piston source that is excited by eight-cycle, 2.5-MHz tone bursts. Axial and lateral field distributions of the superharmonic component and the second harmonic are first calculated based on the nonlinear KZK model and then compared with those experimentally determined at two different source pressures of 0.5 and 1 MPa. Results indicate that the amplitude of the superharmonic component can exceed that of the second harmonic, depending on the axial distance and the fundamental pressure amplitude. Also, the 3-dB beamwidth of the superharmonic component is about 23% narrower than that of the second harmonic. Additional experiments are performed in vitro using liver and fatty tissues in transmission mode and produced two-dimensional images using the fundamental, the second harmonic, and the superharmonic signals. Although the clinical applicability of this work still needs to be assessed, these results indicate that the superharmonic image quality is better than that of the other two images.

Enhanced visible light generation in an active microcavity via third-harmonic conversion beyond the non-depletion approximation

NASA Astrophysics Data System (ADS)

Yu, Rong; Ding, Chunling; Wang, Jiangpeng; Zhang, Duo

2017-12-01

We explore the possibility of using an active doubly resonant microtoroid resonator to produce high-efficiency third-harmonic generation (THG) by exploiting optical third-order nonlinearity. In a microresonator, the active fundamental mode is coherently driven with a continuous-wave input laser at the telecommunication wavelength (1550 nm), and then, the visible THG signal (517 nm) is monitored via an individual bus waveguide. We thoroughly compare our results with those obtained from the conventional passive (i.e., loss) microtoroid resonator by a systematic analysis and detailed numerical simulations based on the Heisenberg-Langevin equations of motion. It is shown that the achievable THG spectrum features an ultralow critical input power. The THG power transmission can be significantly enhanced by about three orders of magnitude at a low input power of 0.1 μ W as compared with the obtained results in the passive microtoroid resonator THG system. Moreover, the THG efficiency can reach up to 100% with optical critical input power as low as a few microwatts. In turn, the analytical expressions of the critical intracavity intensity of the light in the microcavity, the critical input pump power, and the maximum THG efficiency are obtained. The enhanced THG power transmission and high conversion efficiency are attributed to a gain-induced loss compensation in the microtoroid resonator, reducing the effective loss felt by the resonator photons. With state-of-the art technologies in the field of solid-state resonators, including but not limited to microtoroids, the proposed THG scheme is experimentally realizable.

Single cell genomic quantification by non-fluorescence nonlinear microscopy

NASA Astrophysics Data System (ADS)

Kota, Divya; Liu, Jing

2017-02-01

Human epidermal growth receptor 2 (Her2) is a gene which plays a major role in breast cancer development. The quantification of Her2 expression in single cells is limited by several drawbacks in existing fluorescence-based single molecule techniques, such as low signal-to-noise ratio (SNR), strong autofluorescence and background signals from biological components. For rigorous genomic quantification, a robust method of orthogonal detection is highly desirable and we demonstrated it by two non-fluorescent imaging techniques -transient absorption microscopy (TAM) and second harmonic generation (SHG). In TAM, gold nanoparticles (AuNPs) are chosen as an orthogonal probes for detection of single molecules which gives background-free quantifications of single mRNA transcript. In SHG, emission from barium titanium oxide (BTO) nanoprobes was demonstrated which allows stable signal beyond the autofluorescence window. Her2 mRNA was specifically labeled with nanoprobes which are conjugated with antibodies or oligonucleotides and quantified at single copy sensitivity in the cancer cells and tissues. Furthermore, a non-fluorescent super-resolution concept, named as second harmonic super-resolution microscopy (SHaSM), was proposed to quantify individual Her2 transcripts in cancer cells beyond the diffraction limit. These non-fluorescent imaging modalities will provide new dimensions in biomarker quantification at single molecule sensitivity in turbid biological samples, offering a strong cross-platform strategy for clinical monitoring at single cell resolution.

Azimuthally differential pion femtoscopy relative to the second and third harmonic in Pb-Pb 2.76 TeV collision from ALICE

NASA Astrophysics Data System (ADS)

Saleh, Mohammad; Alice Collaboration

2017-11-01

Azimuthally differential femtoscopic measurements, being sensitive to spatio-temporal characteristics of the source as well as to the collective velocity fields at freeze-out, provide very important information on the nature and dynamics of the system evolution. While the HBT radii modulations relative to the second harmonic event plane reflect mostly the spatial geometry of the source, the third harmonic results are mostly defined by the velocity fields [S. A. Voloshin, J. Phys. G38 (2011) 124097. arXiv:arxiv:arXiv:1106.5830, doi:http://dx.doi.org/10.1088/0954-3899/38/12/124097]. Radii variations with respect to the third harmonic event plane unambiguously signal a collective expansion and anisotropy in the flow fields. Event shape engineering (ESE) is a technique proposed to select events corresponding to a particular shape. Azimuthally differential HBT combined with ESE allows for a detailed analysis of the relation between initial geometry, anisotropic flow and the deformation of source shape. We present azimuthally differential pion femtoscopy with respect to second and third harmonic event planes as a function of the pion transverse momentum for different collision centralities in Pb-Pb collisions at √{sNN} = 2.76 TeV. All these results are compared to existing models. The effects of the selection of the events with high elliptic or triangular flow are also presented.

Low-degree gravity change from GPS data of COSMIC and GRACE satellite missions

NASA Astrophysics Data System (ADS)

Lin, Tingjung; Hwang, Cheinway; Tseng, Tzu-Pang; Chao, B. F.

2012-01-01

This paper demonstrates estimation of time-varying gravity harmonic coefficients from GPS data of COSMIC and GRACE satellite missions. The kinematic orbits of COSMIC and GRACE are determined to the cm-level accuracy. The NASA Goddard's GEODYN II software is used to model the orbit dynamics of COSMIC and GRACE, including the effect of a static gravity field. The surface forces are estimated per one orbital period. Residual orbits generated from kinematic and reference orbits serve as observables to determine the harmonic coefficients in the weighted-constraint least-squares. The monthly COSMIC and GRACE GPS data from September 2006 to December 2007 (16 months) are processed to estimate harmonic coefficients to degree 5. The geoid variations from the GPS and CSR RL04 (GRACE) solutions show consistent patterns over space and time, especially in regions of active hydrological changes. The monthly GPS-derived second zonal coefficient closely resembles the SLR-derived and CSR RL04 values, and third and fourth zonal coefficients resemble the CSR RL04 values.

Single-pass high harmonic generation at high repetition rate and photon flux

NASA Astrophysics Data System (ADS)

Hädrich, Steffen; Rothhardt, Jan; Krebs, Manuel; Demmler, Stefan; Klenke, Arno; Tünnermann, Andreas; Limpert, Jens

2016-09-01

Sources of short wavelength radiation with femtosecond to attosecond pulse durations, such as synchrotrons or free electron lasers, have already made possible numerous, and will facilitate more, seminal studies aimed at understanding atomic and molecular processes on fundamental length and time scales. Table-top sources of coherent extreme ultraviolet to soft x-ray radiation enabled by high harmonic generation (HHG) of ultrashort pulse lasers have also gained significant attention in the last few years due to their enormous potential for addressing a plethora of applications, therefore constituting a complementary source to large-scale facilities (synchrotrons and free electron lasers). Ti:sapphire based laser systems have been the workhorses for HHG for decades, but are limited in repetition rate and average power. On the other hand, it has been widely recognized that fostering applications in fields such as photoelectron spectroscopy and microscopy, coincidence detection, coherent diffractive imaging and frequency metrology requires a high repetition rate and high photon flux HHG sources. In this article we will review recent developments in realizing the demanding requirement of producing a high photon flux and repetition rate at the same time. Particular emphasis will be put on suitable ultrashort pulse and high average power lasers, which directly drive harmonic generation without the need for external enhancement cavities. To this end we describe two complementary schemes that have been successfully employed for high power fiber lasers, i.e. optical parametric chirped pulse amplifiers and nonlinear pulse compression. Moreover, the issue of phase-matching in tight focusing geometries will be discussed and connected to recent experiments. We will highlight the latest results in fiber laser driven high harmonic generation that currently produce the highest photon flux of all existing sources. In addition, we demonstrate the first promising applications and discuss the future direction and challenges of this new type of HHG source.

Surface diffusion of Sb on Ge(111) monitored quantitatively with optical second harmonic microscopy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schultz, K.A.; Seebauer, E.G.

Surface diffusion of Sb on Ge(111) has been measured with the newly developed technique of optical second harmonic microscopy. In this method, concentration profiles at submonolayer coverage are imaged directly by surface second harmonic generation with 5 {mu} spatial resolution. A Boltzmann--Matano analysis yields the coverage dependence of the diffusivity {ital D} without parametrization. Experiments were performed at roughly 70% of the bulk melting temperature {ital T}{sub {ital m}}. In the coverage range 0{le}{theta}{le}0.6, the activation energy {ital E}{sub diff} remains constant at 47.5{plus minus}1.5 kcal/mol, but the pre-exponential factor {ital D}{sub 0} decreases from 8.7{times}10{sup 3{plus minus}0.4} to 1.6{times}10{supmore » 2{plus minus}0.4} cm{sup 2}/s. Both {ital E}{sub diff} and {ital D}{sub 0} are quite large, which is consistent with high-temperature measurements in other systems. The inadequacies of current theories for high-temperature surface diffusion are outlined, and a new vacancy model is proposed for low-coverage diffusion. The model accounts semiquantitatively for the large values of {ital E}{sub diff} and {ital D}{sub 0}, and suggests that these quantities may be manipulated using doping levels and photon illumination. An islanding mechanism is proposed to explain the decrease in {ital D}{sub 0} with {theta}.« less

Photo-induced second-order nonlinearity in stoichiometric silicon nitride waveguides

NASA Astrophysics Data System (ADS)

Porcel, Marco A. G.; Mak, Jesse; Taballione, Caterina; Schermerhorn, Victoria K.; Epping, Jörn P.; van der Slot, Peter J. M.; Boller, Klaus-J.

2017-12-01

We report the observation of second-harmonic generation in stoichiometric silicon nitride waveguides grown via low-pressure chemical vapour deposition. Quasi-rectangular waveguides with a large cross section were used, with a height of 1 {\\mu}m and various different widths, from 0.6 to 1.2 {\\mu}m, and with various lengths from 22 to 74 mm. Using a mode-locked laser delivering 6-ps pulses at 1064 nm wavelength with a repetition rate of 20 MHz, 15% of the incoming power was coupled through the waveguide, making maximum average powers of up to 15 mW available in the waveguide. Second-harmonic output was observed with a delay of minutes to several hours after the initial turn-on of pump radiation, showing a fast growth rate between 10$^{-4}$ to 10$^{-2}$ s$^{-1}$, with the shortest delay and highest growth rate at the highest input power. After this first, initial build-up, the second-harmonic became generated instantly with each new turn-on of the pump laser power. Phase matching was found to be present independent of the used waveguide width, although the latter changes the fundamental and second-harmonic phase velocities. We address the presence of a second-order nonlinearity and phase matching, involving an initial, power-dependent build-up, to the coherent photogalvanic effect. The effect, via the third-order nonlinearity and multiphoton absorption leads to a spatially patterned charge separation, which generates a spatially periodic, semi-permanent, DC-field-induced second-order susceptibility with a period that is appropriate for quasi-phase matching. The maximum measured second-harmonic conversion efficiency amounts to 0.4% in a waveguide with 0.9 x 1 {\\mu}m$^2$ cross section and 36 mm length, corresponding to 53 {\\mu}W at 532 nm with 13 mW of IR input coupled into the waveguide. The according $\\chi^{(2)}$ amounts to 3.7 pm/V, as retrieved from the measured conversion efficiency.

Optical Manifestations of the Electron-Electron Interaction

NASA Astrophysics Data System (ADS)

Portengen, Taco

1995-01-01

In this thesis, two optical manifestations of the electron-electron interaction are studied: the Fermi -edge singularity in doped quantum wells and quantum wires, and second-harmonic generation in mixed-valent compounds. First, we construct a theory of the Fermi-edge singularity that can systematically account for the finite mass of a hole created in the valence subband of a quantum well or quantum wire. The dynamical response for finite hole mass depends crucially on the dimensionality of the Fermi sea. Whereas in three dimensions the infrared divergence is suppressed, in two dimensions a one-over-square-root singularity survives, while in one dimension the spectrum is even more singular with recoil than without recoil. This explains the large optical singularities observed in quantum wires. Correlations change the prefactor, but not the exponent of the threshold behaviour in two and in three dimensions, while in one dimension, they affect neither the prefactor nor the exponent. Second, we apply our theory to the Frohlich polaron, a manifestation of the electron-phonon rather than the electron-electron interaction. The new method of calculating the Green's function removes unphysical features of the conventional cumulant expansion that had remained unnoticed in the literature up to now. Third, in an effort to investigate the impact of coherence on optical properties, we calculate the linear and nonlinear optical characteristics of mixed-valent compounds. Second -harmonic generation can only occur for solutions of the theoretical Falicov-Kimball model that have a built-in coherence between the itinerant d-electrons and localized f-holes. By contrast, second-harmonic generation cannot occur for solutions with f-site occupation as a good quantum number. The interaction between optically created quasiparticles leads to a threshold singularity in the absorption spectrum, and greatly enhances the second-harmonic conversion efficiency at half the gap frequency. As an experimental test of coherence we propose the measurement of the second-harmonic susceptibility of SmB_6..

Synchronous digitization for high dynamic range lock-in amplification in beam-scanning microscopy

PubMed Central

Muir, Ryan D.; Sullivan, Shane Z.; Oglesbee, Robert A.; Simpson, Garth J.

2014-01-01

Digital lock-in amplification (LIA) with synchronous digitization (SD) is shown to provide significant signal to noise (S/N) and linear dynamic range advantages in beam-scanning microscopy measurements using pulsed laser sources. Direct comparisons between SD-LIA and conventional LIA in homodyne second harmonic generation measurements resulted in S/N enhancements consistent with theoretical models. SD-LIA provided notably larger S/N enhancements in the limit of low light intensities, through the smooth transition between photon counting and signal averaging developed in previous work. Rapid beam scanning instrumentation with up to video rate acquisition speeds minimized photo-induced sample damage. The corresponding increased allowance for higher laser power without sample damage is advantageous for increasing the observed signal content. PMID:24689588

Characterization of a reflective objective with multiphoton microscopy

NASA Astrophysics Data System (ADS)

Kabir, Mohammad M.; Choubal, Aakash M.; Sivaguru, Mayandi; Toussaint, Kimani C.

2018-02-01

Reflective objectives (ROs) can reduce chromatic aberration across a wide wavelength range in multiphoton microscopy (MPM). However, a systematic characterization of the performance of ROs has not been carried out. In this paper, we analyze the performance of a 0.5 numerical-aperture (NA) RO and compare it with a 0.55 NA standard glass objective (SO), using two-photon fluorescence (TPF) and second-harmonic generation (SHG). For experiments extending 1 octave in visible and NIR wavelengths, the SO introduces defocusing errors of 25% for TPF images of sub-diffraction fluorescent beads and 10% for SHG images of collagen fibers. For both imaging systems, the RO provides a corresponding error of 4%. This work highlights the potential usefulness of ROs for multimodal MPM applications.

Multimodal optical workstation for simultaneous linear, nonlinear microscopy and nanomanipulation: upgrading a commercial confocal inverted microscope.

PubMed

Mathew, Manoj; Santos, Susana I C O; Zalvidea, Dobryna; Loza-Alvarez, Pablo

2009-07-01

In this work we propose and build a multimodal optical workstation that extends a commercially available confocal microscope (Nikon Confocal C1-Si) to include nonlinear/multiphoton microscopy and optical manipulation/stimulation tools such as nanosurgery. The setup allows both subsystems (confocal and nonlinear) to work independently and simultaneously. The workstation enables, for instance, nanosurgery along with simultaneous confocal and brightfield imaging. The nonlinear microscopy capabilities are added around the commercial confocal microscope by exploiting all the flexibility offered by this microscope and without need for any mechanical or electronic modification of the confocal microscope systems. As an example, the standard differential interference contrast condenser and diascopic detector in the confocal microscope are readily used as a forward detection mount for second harmonic generation imaging. The various capabilities of this workstation, as applied directly to biology, are demonstrated using the model organism Caenorhabditis elegans.

Utilizing nonlinear optical microscopy to investigate the development of early cancer in nude mice in vivo

NASA Astrophysics Data System (ADS)

Wang, Chun-Chin; Li, Feng-Chieh; Lin, Sung-Jan; Lo, Wen; Dong, Chen-Yuan

2007-07-01

In this investigation, we used in vivo nonlinear optical microscopy to image normal and carcinogen DMBA treated skin tissues of nude mice. We acquired two-photon autofluroescence and second harmonic generation (SHG) images of the skin tissue, and applied the ASI (Autofluorescence versus SHG Index) to the resulting image. This allows us to visualize and quantify the interaction between mouse skin cells and the surrounding connective tissue. We found that as the imaging depth increases, ASI has a different distribution in the normal and the treated skin tissues. Since the DMBA treated skin eventually became squamous cell carcinoma (SCC), our results show that the physiological changes to mouse skin en route to become cancer can be effectively tracked by multiphoton microscopy. We envision this approach to be effective in studying tumor biology and tumor treatment procedures.

The collagen structure of equine articular cartilage characterized using polarization-sensitive optical coherence tomography and non-linear microscopy

NASA Astrophysics Data System (ADS)

Mansfield, Jessica C.; Ugryumova, Nadya; Knapp, Karen M.; Matcher, Stephen J.

2006-09-01

Equine articular cartilage has been imaged using both polarization-sensitive optical coherence tomography (PS-OCT) and non-linear microscopy. PS-OCT has been used to spatially map the birefringence in the cartilage and we have found that in the vicinity of the lesion the images display a characteristic disruption in the regular birefringence bands shown by normal cartilage. We also note that significant (e.g. x2) variations in the apparent birefringence of samples taken from young (18 month) animals that otherwise appear visually homogeneous are found over spatial scales of a few millimeters. We have also imaged the cartilage using non-linear microscopy and compare the scans taken with second harmonic generation (SHG) light and the two photon fluorescence (TPF) light. SHG images collected using 800 nm excitation reveals the spatial distribution of collagen fibers, whilst TPF images clearly shows the distribution of intracellular and pericellular fluorophores.

Assessment of fibrotic liver disease with multimodal nonlinear optical microscopy

NASA Astrophysics Data System (ADS)

Lu, Fake; Zheng, Wei; Tai, Dean C. S.; Lin, Jian; Yu, Hanry; Huang, Zhiwei

2010-02-01

Liver fibrosis is the excessive accumulation of extracellular matrix proteins such as collagens, which may result in cirrhosis, liver failure, and portal hypertension. In this study, we apply a multimodal nonlinear optical microscopy platform developed to investigate the fibrotic liver diseases in rat models established by performing bile duct ligation (BDL) surgery. The three nonlinear microscopy imaging modalities are implemented on the same sectioned tissues of diseased model sequentially: i.e., second harmonic generation (SHG) imaging quantifies the contents of the collagens, the two-photon excitation fluorescence (TPEF) imaging reveals the morphology of hepatic cells, while coherent anti-Stokes Raman scattering (CARS) imaging maps the distributions of fats or lipids quantitatively across the tissue. Our imaging results show that during the development of liver fibrosis (collagens) in BDL model, fatty liver disease also occurs. The aggregated concentrations of collagen and fat constituents in liver fibrosis model show a certain correlationship between each other.

The development of novel Ytterbium fiber lasers and their applications

NASA Astrophysics Data System (ADS)

Nie, Bai

The aim of my Ph.D. research is to push the fundamental limits holding back the development of novel Yb fiber lasers with high pulse energy and short pulse duration. The purpose of developing these lasers is to use them for important applications such as multiphoton microscopy and laser-induced breakdown spectroscopy. My first project was to develop a short-pulse high-energy ultrafast fiber laser for multiphoton microscopy. To achieve high multiphoton efficiency and depth resolved tissue imaging, ultrashort pulse duration and high pulse energy are required. In order to achieve this, an all-normal dispersion cavity design was adopted. Output performances of the built lasers were investigated by varying several cavity parameters, such as pump laser power, fiber length and intra-cavity spectral filter bandwidth. It was found that the length of the fiber preceding the gain fiber is critical to the laser performance. Generally, the shorter the fiber is, the broader the output spectrum is. The more interesting parameter is the intra-cavity spectral filter bandwidth. Counter intuitively, laser cavities using narrower bandwidth spectral filters generated much broader spectra. It was also found that fiber lasers with very narrow spectral filters produced laser pulses with parabolic profile, which are referred to as self-similar pulses or similaritons. This type of pulse can avoid wave-breaking and is an optimal approach to generate pulses with high pulse energy and ultrashort pulse duration. With a 3nm intra-cavity spectral filter, output pulses with about 20 nJ pulse energy were produced and compressed to about 41 fs full-width-at-half-maximum (FWHM) pulse duration. Due to the loss in the compression device, the peak power of the compressed pulses is about 250 kW. It was the highest peak power generated from a fiber oscillator when this work was published. This laser was used for multiphoton microscopy on living tissues like Drosophila larva and fruit fly wings. Several imaging methods, such as two-photon-excited fluorescence, second harmonic generation, and third harmonic generation, were performed. Not only were single layers of thin tissue imaged, but also depth resolved imaging of thick samples was tested, and three-dimensional image reconstruction was demonstrated. The other project was to develop a simple fiber oscillator for laser-induced breakdown spectroscopy (LIBS). Laser pulses with high energy, high ablation efficiency and low ablation threshold are desirable for this application. We built a fiber laser using up to 200 m long fiber and scaled the output pulse energy up to 450 nJ. This laser was operated in an unusual mode-locking regime and produced noise-like pulses, which have a picosecond long pulse envelope containing multiple irregular femtosecond sub-pulses. This type of pulse was mostly ignored by many earlier researchers. Intra-cavity spectral filters did not affect the laser performance as much as in the similariton lasers and were removed from the laser cavity. Characteristics of our noise-like laser, such as MHz repetition rate, broad spectrum, and picosecond-long pulse envelope containing multiple femtosecond sub-pulses, were found to meet the requirement of an ideal laser source for LIBS. A simple LIBS setup using our laser was demonstrated and atomic emission spectra with very good signal-to-noise ratio were obtained. Composition detection, qualitative concentration determination, and trace detection were also tested. These tests show that our noise-like fiber laser is an ideal laser source for a low-cost and portable LIBS system.

Damage Characterization and Real-Time Health Monitoring of Aerospace Materials Using Innovative NDE Tools

NASA Astrophysics Data System (ADS)

Matikas, Theodore E.

2010-07-01

The objective of this work is to characterize the damage and monitor in real-time aging structural components used in aerospace applications by means of advanced nondestructive evaluation techniques. Two novel experimental methodologies are used in this study, based on ultrasonic microscopy and nonlinear acoustics. It is demonstrated in this work that ultrasonic microscopy can be successfully utilized for local elastic property measurement, crack-size determination as well as for interfacial damage evaluation in high-temperature materials, such as metal matrix composites. Nonlinear acoustics enables real-time monitoring of material degradation in aerospace structures. When a sinusoidal ultrasonic wave of a given frequency and of sufficient amplitude is introduced into a nonharmonic solid, the fundamental wave distorts as it propagates, and therefore the second and higher harmonics of the fundamental frequency are generated. Measurements of the amplitude of these harmonics provide information on the coefficient of second- and higher-order terms of the stress-strain relation for a nonlinear solid. It is shown in this article that the material bulk nonlinear parameter for metallic alloy samples at different fatigue levels exhibits large changes compared to linear ultrasonic parameters, such as velocity and attenuation.

Ultra-broadband ptychography with self-consistent coherence estimation from a high harmonic source

NASA Astrophysics Data System (ADS)

Odstrčil, M.; Baksh, P.; Kim, H.; Boden, S. A.; Brocklesby, W. S.; Frey, J. G.

2015-09-01

With the aim of improving imaging using table-top extreme ultraviolet sources, we demonstrate coherent diffraction imaging (CDI) with relative bandwidth of 20%. The coherence properties of the illumination probe are identified using the same imaging setup. The presented methods allows for the use of fewer monochromating optics, obtaining higher flux at the sample and thus reach higher resolution or shorter exposure time. This is important in the case of ptychography when a large number of diffraction patterns need to be collected. Our microscopy setup was tested on a reconstruction of an extended sample to show the quality of the reconstruction. We show that high harmonic generation based EUV tabletop microscope can provide reconstruction of samples with a large field of view and high resolution without additional prior knowledge about the sample or illumination.

Stimulated Brillouin scattering during electron gyro-harmonic heating at EISCAT

NASA Astrophysics Data System (ADS)

Fu, H. Y.; Scales, W. A.; Bernhardt, P. A.; Briczinski, S. J.; Kosch, M. J.; Senior, A.; Rietveld, M. T.; Yeoman, T. K.; Ruohoniemi, J. M.

2015-08-01

Observations of secondary radiation, stimulated electromagnetic emission (SEE), produced during ionospheric modification experiments using ground-based, high-power, high-frequency (HF) radio waves are considered. The High Frequency Active Auroral Research Program (HAARP) facility is capable of generating narrowband SEE in the form of stimulated Brillouin scatter (SBS) and stimulated ion Bernstein scatter (SIBS) in the SEE spectrum. Such narrowband SEE spectral lines have not been reported using the European Incoherent Scatter (EISCAT) heater facility before. This work reports the first EISCAT results of narrowband SEE spectra and compares them to SEE previously observed at HAARP during electron gyro-harmonic heating. An analysis of experimental SEE data shows observations of emission lines within 100 Hz of the pump frequency, interpreted as SBS, during the 2012 July EISCAT campaign. Experimental results indicate that SBS strengthens as the pump frequency approaches the third electron gyro-harmonic. Also, for different heater antenna beam angles, the CUTLASS radar backscatter induced by HF radio pumping is suppressed near electron gyro-harmonics, whereas electron temperature enhancement weakens as measured by EISCAT/UHF radar. The main features of these new narrowband EISCAT observations are generally consistent with previous SBS measurements at HAARP.

CAVE: the design of a precision metrology instrument for studying performance of KDP crystals

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hibbard, R.L., LLNL

1998-03-30

A device has been developed to measure the frequency conversion performance of large aperture potassium dihydrogen phosphate (KDP) crystals. Third harmonic generation using ICDP is critical to the function of the National Ignition Facility (NIF) laser. The crystals in the converter can be angularly or thermally tuned but are subject to larger aperture inhomogeneities that are functions of growth manufacturing and - mounting. The CAVE (Crystal Alignment Verification Equipment) instrument scans the crystals in a thermally and mechanically controlled environment to determine the local peak tuning angles. The CAVE can then estimate the optimum tuning angle and conversion efficiency overmore » the entire aperture. Coupled with other metrology techniques, the CAVE will help determine which crystal life-cycle components most affect harmonic conversion.« less

Modelling nonlinearity in superconducting split ring resonator and its effects on metamaterial structures

NASA Astrophysics Data System (ADS)

Mazdouri, Behnam; Mohammad Hassan Javadzadeh, S.

2017-09-01

Superconducting materials are intrinsically nonlinear, because of nonlinear Meissner effect (NLME). Considering nonlinear behaviors, such as harmonic generation and intermodulation distortion (IMD) in superconducting structures, are very important. In this paper, we proposed distributed nonlinear circuit model for superconducting split ring resonators (SSRRs). This model can be analyzed by using Harmonic Balance method (HB) as a nonlinear solver. Thereafter, we considered a superconducting metamaterial filter which was based on split ring resonators and we calculated fundamental and third-order IMD signals. There are good agreement between nonlinear results from proposed model and measured ones. Additionally, based on the proposed nonlinear model and by using a novel method, we considered nonlinear effects on main parameters in the superconducting metamaterial structures such as phase constant (β) and attenuation factor (α).

Investigation of the spatial distribution of second-order nonlinearity in thermally poled optical fibers.

PubMed

An, Honglin; Fleming, Simon

2005-05-02

The spatial distribution of second-order nonlinearity in thermally poled optical fibers was characterized by second-harmonic microscopy. The second-order nonlinearity was found to be confined to a thin layer close to the anode surface and progressed further into the silica as the poling time increased. Position uncertainty of the anode metal wire was observed to have an effect, as the nonlinear layers were found not always symmetrically located around the nearest points between the anode and cathode. Optical microscopy results were obtained on etched poled fiber cross-sections and compared with those from second-harmonic microscopy.

Physical Chemical Studies on Molecular Composite Compositions

DTIC Science & Technology

1993-02-20

be known over a wide range of wavelengths, from UV-Visible to near infrared , for second and third harmonic generation (SHG and THG) techniques. Here...visible to near infrared radiations. The measured birefringence is high, e.g., I nE - no I varies from 0.05 to 0.09 at 632.8 nm for polymer...indices no and nE, respectively, of nematic solutions of the rodlike poly(p-phenylene benzobisthiazole), PBT, for wavelengths from the visible to near

Odd Harmonics in Exoplanet Photometry: Weather or Artifact?

NASA Astrophysics Data System (ADS)

Chayes, Victoria; Cowan, Nicholas; Bouffard, Élie; Haggard, Hal

2017-01-01

In the Fourier decomposition of light curves of exoplanets observed by the Kepler mission, one expects to see power in the first mode, from the planet orbiting the star, and the second mode, from ellipsoidal variations. Observations of power in the third mode of planets such as HAT-P-7b and Kepler-13Ab are as of yet unexplained. Using a spherical harmonic basis we analyze planet maps to find their corresponding light curves and show that no planet observed edge-on can produce these third harmonics with either reflected light or thermal emissions. Further numerical and analytic calculations put upper bounds on the power in the third mode that can be produced by planets not transiting perfectly edge-on, or with time-variable maps. We find the expected order of magnitude of these contributions to be at most two orders of magnitude below the first harmonic. The North-South asymmetric features or time-variable maps that could produce such harmonics would suggest exoplanetary weather if observed. However, more careful analysis of tidal effects on the stars of HAT-P-7b and Kepler-13Ab suggest that these particular harmonics are stellar in origin. Élie Bouffard was supported by an iREx summer internship. We thank the International Space Science Institute in Bern, Switzerland, for hosting the Exo-Cartography workshop series.

Wire bonding quality monitoring via refining process of electrical signal from ultrasonic generator

NASA Astrophysics Data System (ADS)

Feng, Wuwei; Meng, Qingfeng; Xie, Youbo; Fan, Hong

2011-04-01

In this paper, a technique for on-line quality detection of ultrasonic wire bonding is developed. The electrical signals from the ultrasonic generator supply, namely, voltage and current, are picked up by a measuring circuit and transformed into digital signals by a data acquisition system. A new feature extraction method is presented to characterize the transient property of the electrical signals and further evaluate the bond quality. The method includes three steps. First, the captured voltage and current are filtered by digital bandpass filter banks to obtain the corresponding subband signals such as fundamental signal, second harmonic, and third harmonic. Second, each subband envelope is obtained using the Hilbert transform for further feature extraction. Third, the subband envelopes are, respectively, separated into three phases, namely, envelope rising, stable, and damping phases, to extract the tiny waveform changes. The different waveform features are extracted from each phase of these subband envelopes. The principal components analysis (PCA) method is used for the feature selection in order to remove the relevant information and reduce the dimension of original feature variables. Using the selected features as inputs, an artificial neural network (ANN) is constructed to identify the complex bond fault pattern. By analyzing experimental data with the proposed feature extraction method and neural network, the results demonstrate the advantages of the proposed feature extraction method and the constructed artificial neural network in detecting and identifying bond quality.

Low- and high-order harmonic generation in the extended plasmas produced by laser ablation of zinc and manganese targets

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ganeev, R. A., E-mail: rashid-ganeev@mail.ru; Physical Department, Voronezh State University, Voronezh 394006; Baba, M.

The systematic studies of the harmonic generation of ultrashort laser pulses in the 5-mm-long Zn and Mn plasmas (i.e., application of nanosecond, picosecond, and femtosecond pulses for ablation, comparison of harmonic generation from atomic, ionic, and cluster-contained species of plasma, variation of plasma length, two-color pump of plasmas, etc.) are presented. The conversion efficiency of the 11th–19th harmonics generated in the Zn plasma was ∼5 × 10{sup −5}. The role of the ionic resonances of Zn near the 9th and 10th harmonics on the enhancement of harmonics is discussed. The enhancement of harmonics was also analyzed using the two-color pump of extendedmore » plasmas, which showed similar intensities of the odd and even harmonics along the whole range of generation. The harmonics up to the 107th order were demonstrated in the case of manganese plasma. The comparison of harmonic generation in the 5-mm-long and commonly used short (≤0.5 mm) plasma plumes showed the advanced properties of extended media.« less

Observation of tendon repair in animal model using second-harmonic-generation microscopy

NASA Astrophysics Data System (ADS)

Hase, Eiji; Minamikawa, Takeo; Sato, Katsuya; Takahashi, Mitsuhiko; Yasui, Takashi

2016-03-01

Tendon rupture is a trauma difficult to recover the condition before injury. In previous researches, tensile test and staining method have been widely used to elucidate the mechanism of the repair process from the viewpoints of the mechanical property and the histological findings. However, since both methods are destructive and invasive, it is difficult to obtain both of them for the same sample. If both the mechanical property and the histological findings can be obtained from the same sample, one may obtain new findings regarding mechanisms of tendon repairing process. In this paper, we used second-harmonic-generation (SHG) microscopy, showing high selectivity and good image contrast to collagen molecules as well as high spatial resolution, optical three-dimensional sectioning, deep penetration, and without additional staining. Since SHG light intensity sensitively reflects the structural maturity of collagen molecule and its aggregates, it will be a good indicator for the repairing degree of the ruptured tendon. From comparison of SHG images between the 4-weeks-repaired tendon and the sound tendon in the animal model, we confirmed that SHG light intensity of the repaired tendon was significantly lower than that of the sound tendon, indicating that the collagen structure in the repaired tendon is still immature. Furthermore, we performed both SHG imaging and the tensile test for the same sample, and confirmed a correlation between them. This result shows a potential of SHG light for an indicator of the histological and mechanical recovery of the ruptured tendon.

Preparation, linear and NLO properties of DNA-CTMA-SBE complexes

NASA Astrophysics Data System (ADS)

Manea, Ana-Maria; Rau, Ileana; Kajzar, Francois; Meghea, Aurelia

2013-10-01

Synthesis of deoxyribonucleic acid (DNA) - was cetyltrimethylammonium (CTMA) - sea buckthorn extract (SBE) at different concentrations is decribed. The complexes were processed into good optical quality thin films by spin coating on different substrates such as: glass, silica and ITO covered glass substrates. SBE contains many bioactive substances that can be used in the treatment of several diseases, such as cardiovascular disease, cancer, and acute mountain sickness. The obtained thin films were characterized for their spectroscopic, fluorescent, linear and nonlinear optical properties as function of SBE concentration. The third-order nonlinear optical (NLO) properties of thin films were determined by the optical third-harmonic generation technique at 1 064.2 nm fundamental wavelength.

Effect of halogenation on the nonlinear optical properties of porthyrin and substituted porphyrins

NASA Technical Reports Server (NTRS)

Cardelino, Beatriz H.; Moore, Craig E.; Benloss, Angela; Thompson, Albert N., Jr.; Richards, Rosalie A.; Roney, Celeste A.; Sanghadasa, Mohan

1998-01-01

The effect that fluorine and chlorine substitution has on the nonlinear optical properties of porphyrin, tetramethylporphyrin and tetraphenylporphyrin has been theoretically studied. The calculations of nonlinear optical properties have been obtained by performing finite-field calculations on structures determined by semiempirical methods. In addition, tetra(p-chlorophenyl)porphyrin and tetra(p-bromophenyl)porphyrin were synthesized by the condensation of pyrrol and the appropriate aldehyde. Thin films of polymethylmethacrylate were obtained containing these materials, by spin coating onto glass substrates. The films were characterized by third-harmonic generation. It was determined that the experimental conditions enhance the third-order polarizability of the tetraphenylporphyrins by a factor of about 1.6.

Invited Review Article: Pump-probe microscopy.

PubMed

Fischer, Martin C; Wilson, Jesse W; Robles, Francisco E; Warren, Warren S

2016-03-01

Multiphoton microscopy has rapidly gained popularity in biomedical imaging and materials science because of its ability to provide three-dimensional images at high spatial and temporal resolution even in optically scattering environments. Currently the majority of commercial and home-built devices are based on two-photon fluorescence and harmonic generation contrast. These two contrast mechanisms are relatively easy to measure but can access only a limited range of endogenous targets. Recent developments in fast laser pulse generation, pulse shaping, and detection technology have made accessible a wide range of optical contrasts that utilize multiple pulses of different colors. Molecular excitation with multiple pulses offers a large number of adjustable parameters. For example, in two-pulse pump-probe microscopy, one can vary the wavelength of each excitation pulse, the detection wavelength, the timing between the excitation pulses, and the detection gating window after excitation. Such a large parameter space can provide much greater molecular specificity than existing single-color techniques and allow for structural and functional imaging without the need for exogenous dyes and labels, which might interfere with the system under study. In this review, we provide a tutorial overview, covering principles of pump-probe microscopy and experimental setup, challenges associated with signal detection and data processing, and an overview of applications.

Invited Review Article: Pump-probe microscopy

PubMed Central

Wilson, Jesse W.; Robles, Francisco E.; Warren, Warren S.

2016-01-01

Multiphoton microscopy has rapidly gained popularity in biomedical imaging and materials science because of its ability to provide three-dimensional images at high spatial and temporal resolution even in optically scattering environments. Currently the majority of commercial and home-built devices are based on two-photon fluorescence and harmonic generation contrast. These two contrast mechanisms are relatively easy to measure but can access only a limited range of endogenous targets. Recent developments in fast laser pulse generation, pulse shaping, and detection technology have made accessible a wide range of optical contrasts that utilize multiple pulses of different colors. Molecular excitation with multiple pulses offers a large number of adjustable parameters. For example, in two-pulse pump-probe microscopy, one can vary the wavelength of each excitation pulse, the detection wavelength, the timing between the excitation pulses, and the detection gating window after excitation. Such a large parameter space can provide much greater molecular specificity than existing single-color techniques and allow for structural and functional imaging without the need for exogenous dyes and labels, which might interfere with the system under study. In this review, we provide a tutorial overview, covering principles of pump-probe microscopy and experimental setup, challenges associated with signal detection and data processing, and an overview of applications. PMID:27036751

Ultrasound contrast agent imaging: Real-time imaging of the superharmonics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Peruzzini, D.; Viti, J.; Erasmus MC, ’s-Gravendijkwal 230, Faculty Building, Ee 2302, 3015 CE Rotterdam

2015-10-28

Currently, in medical ultrasound contrast agent (UCA) imaging the second harmonic scattering of the microbubbles is regularly used. This scattering is in competition with the signal that is caused by nonlinear wave propagation in tissue. It was reported that UCA imaging based on the third or higher harmonics, i.e. “superharmonic” imaging, shows better contrast. However, the superharmonic scattering has a lower signal level compared to e.g. second harmonic signals. This study investigates the contrast-to-tissue ratio (CTR) and signal to noise ratio (SNR) of superharmonic UCA scattering in a tissue/vessel mimicking phantom using a real-time clinical scanner. Numerical simulations were performedmore » to estimate the level of harmonics generated by the microbubbles. Data were acquired with a custom built dual-frequency cardiac phased array probe. Fundamental real-time images were produced while beam formed radiofrequency (RF) data was stored for further offline processing. The phantom consisted of a cavity filled with UCA surrounded by tissue mimicking material. The acoustic pressure in the cavity of the phantom was 110 kPa (MI = 0.11) ensuring non-destructivity of UCA. After processing of the acquired data from the phantom, the UCA-filled cavity could be clearly observed in the images, while tissue signals were suppressed at or below the noise floor. The measured CTR values were 36 dB, >38 dB, and >32 dB, for the second, third, and fourth harmonic respectively, which were in agreement with those reported earlier for preliminary contrast superharmonic imaging. The single frame SNR values (in which ‘signal’ denotes the signal level from the UCA area) were 23 dB, 18 dB, and 11 dB, respectively. This indicates that noise, and not the tissue signal, is the limiting factor for the UCA detection when using the superharmonics in nondestructive mode.« less

Two-Color Laser High-Harmonic Generation in Cavitated Plasma Wakefields

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schroeder, Carl; Benedetti, Carlo; Esarey, Eric

2016-10-03

A method is proposed for producing coherent x-rays via high-harmonic generation using a laser interacting with highly-stripped ions in cavitated plasma wakefields. Two laser pulses of different colors are employed: a long-wavelength pulse for cavitation and a short-wavelength pulse for harmonic generation. This method enables efficient laser harmonic generation in the sub-nm wavelength regime.

Non-linear imaging techniques visualize the lipid profile of C. elegans

NASA Astrophysics Data System (ADS)

Mari, Meropi; Petanidou, Barbara; Palikaras, Konstantinos; Fotakis, Costas; Tavernarakis, Nektarios; Filippidis, George

2015-07-01

The non-linear techniques Second and Third Harmonic Generation (SHG, THG) have been employed simultaneously to record three dimensional (3D) imaging and localize the lipid content of the muscular areas (ectopic fat) of Caenorhabditis elegans (C. elegans). Simultaneously, Two-Photon Fluorescence (TPEF) was used initially to localize the stained lipids with Nile Red, but also to confirm the THG potential to image lipids successfully. In addition, GFP labelling of the somatic muscles, proves the initial suggestion of the existence of ectopic fat on the muscles and provides complementary information to the SHG imaging of the pharynx. The ectopic fat may be related to a complex of pathological conditions including type-2 diabetes, hypertension and cardiovascular diseases. The elucidation of the molecular path leading to the development of metabolic syndrome is a vital issue with high biological significance and necessitates accurate methods competent of monitoring lipid storage distribution and dynamics in vivo. THG microscopy was employed as a quantitative tool to monitor the lipid accumulation in non-adipose tissues in the pharyngeal muscles of 12 unstained specimens while the SHG imaging revealed the anatomical structure of the muscles. The ectopic fat accumulation on the pharyngeal muscles increases in wild type (N2) C. elegans between 1 and 9 days of adulthood. This suggests a correlation of the ectopic fat accumulation with the aging. Our results can provide new evidence relating the deposition of ectopic fat with aging, but also validate SHG and THG microscopy modalities as new, non-invasive tools capable of localizing and quantifying selectively lipid accumulation and distribution.

Texture analysis applied to second harmonic generation image data for disease classification and development of a multi-view second harmonic generation imaging platform

NASA Astrophysics Data System (ADS)

Wen, Lianggong

Many diseases, e.g. ovarian cancer, breast cancer and pulmonary fibrosis, are commonly associated with drastic alterations in surrounding connective tissue, and changes in the extracellular matrix (ECM) are associated with the vast majority of cellular processes in disease progression and carcinogenesis: cell differentiation, proliferation, biosynthetic ability, polarity, and motility. We use second harmonic generation (SHG) microscopy for imaging the ECM because it is a non-invasive, non-linear laser scanning technique with high sensitivity and specificity for visualizing fibrillar collagen. In this thesis, we are interested in developing imaging techniques to understand how the ECM, especially the collagen architecture, is remodeled in diseases. To quantitate remodeling, we implement a 3D texture analysis to delineate the collagen fibrillar morphology observed in SHG microscopy images of human normal and high grade malignant ovarian tissues. In the learning stage, a dictionary of "textons"---frequently occurring texture features that are identified by measuring the image response to a filter bank of various shapes, sizes, and orientations---is created. By calculating a representative model based on the texton distribution for each tissue type using a training set of respective mages, we then perform classification between normal and high grade malignant ovarian tissues classification based on the area under receiver operating characteristic curves (true positives versus false positives). The local analysis algorithm is a more general method to probe rapidly changing fibrillar morphologies than global analyses such as FFT. It is also more versatile than other texture approaches as the filter bank can be highly tailored to specific applications (e.g., different disease states) by creating customized libraries based on common image features. Further, we describe the development of a multi-view 3D SHG imaging platform. Unlike fluorescence microscopy, SHG excites intrinsic characteristics of collagen, bypassing the need for additional primary and secondary imaging labels. However, single view image collection from endogenous SHG contrast of collagen molecules is not "a true 3D technique", because collagen fibers oriented along the plane of the lasers used to excite them are invisible to the excitation The loss of information means that researchers cannot resolve the 3D structure of the ECM using this technique. We are developing a new, multi-view approach that involves rotation of agarose embedded sample in FEP tubing, so that the excitation beam path travels to from multiple angles, to reveal new insight in understanding the 3D collagen structure and its role in normal and diseased tissue.

Frequency-resolved optical gating with the use of second-harmonic generation

DOE Office of Scientific and Technical Information (OSTI.GOV)

DeLong, K.W.; Trebino, R.; Hunter, J.

1994-11-01

We discuss the use of second-harmonic generation (SHG) as the nonlinearity in the technique of frequency-resolved optical gating (FROG) for measuring the full intensity and phase evolution of an arbitrary ultrashort pulse. FROG that uses a third-order nonlinearity in the polarization-gate geometry has proved extremely successful, and the algorithm required for extraction of the intensity and the phase from the experimental data is quite robust. However, for pulse intensities less than [similar to] 1 MW, third-order nonlinearities generate insufficient signal strength, and therefore SHG FROG appears necessary. We discuss the theoretical, algorithmic, and experimental considerations of SHG FROG in detail.more » SHG FROG has an ambiguity in the direction of time, and its traces are somewhat unintuitive. Also, previously published algorithms are generally ineffective at extracting the intensity and the phase of an arbitrary laser pulse from the SHG FROG trace. We present an improved pulse-retrieval algorithm, based on the method of generalized projections, that is far superior to the previously published algorithms, although it is still not so robust as the polarization-gate algorithm. We discuss experimental sources of error such as pump depletion and group-velocity mismatch. We also present several experimental examples of pulses measured with SHG FROG and show that the derived intensities and phases are in agreement with more conventional diagnostic techniques, and we demonstrate the high-dynamic-range capability of SHG FROG. We conclude that, despite the above drawbacks, SHG FROG should be useful in measuring low-energy pulses.« less

Harmonics distribution of iron oxide nanoparticles solutions under diamagnetic background

NASA Astrophysics Data System (ADS)

Saari, Mohd Mawardi; Che Lah, Nurul Akmal; Sakai, Kenji; Kiwa, Toshihiko; Tsukada, Keiji

2018-04-01

The static and dynamic magnetizations of low concentrated multi-core iron oxide nanoparticles solutions were investigated by a specially developed high-Tc Superconducting Quantum Interference Device (SQUID) magnetometer. The size distribution of iron oxide cores was determined from static magnetization curves concerning different concentrations. The simulated harmonics distribution was compared to the experimental results. Effect of the diamagnetic background from carrier liquid to harmonics distribution was investigated with respect to different intensity and position of peaks in the magnetic moment distribution using a numerical simulation. It was found that the diamagnetic background from carrier liquid of iron oxide nanoparticles affected the harmonics distribution as their concentration decreased and depending on their magnetic moment distribution. The first harmonic component was susceptible to the diamagnetic contribution of carrier liquid when the concentration was lower than 24 μg/ml. The second and third harmonics were affected when the peak position of magnetic moment distribution was smaller than m = 10-19 Am2 and the concentration was 10 ng/ml. A highly sensitive detection up to sub-nanogram of iron oxide nanoparticles in solutions can be achieved by utilizing second and third harmonic components.

Third harmonic ac susceptibility of superconductors with finite thickness

NASA Astrophysics Data System (ADS)

Qin, M. J.; Ong, C. K.

Third harmonic ac susceptibility of superconducting strips with finite thickness in perpendicularly applied magnetic field Ha = H0 sin(ω t) have been calculated. The flux creep effect is taken into account by using a power-law electric field E( j) = Ec( j/ jc) n. Results for different thicknesses and creep exponents n have been derived and compared to the results derived from the Bean critical state model.

Methods, systems and apparatus for optimization of third harmonic current injection in a multi-phase machine

DOEpatents

Gallegos-Lopez, Gabriel

2012-10-02

Methods, system and apparatus are provided for increasing voltage utilization in a five-phase vector controlled machine drive system that employs third harmonic current injection to increase torque and power output by a five-phase machine. To do so, a fundamental current angle of a fundamental current vector is optimized for each particular torque-speed of operating point of the five-phase machine.

Tailored semiconductors for high-harmonic optoelectronics

NASA Astrophysics Data System (ADS)

Sivis, Murat; Taucer, Marco; Vampa, Giulio; Johnston, Kyle; Staudte, André; Naumov, Andrei Yu.; Villeneuve, D. M.; Ropers, Claus; Corkum, P. B.

2017-07-01

The advent of high-harmonic generation in gases 30 years ago set the foundation for attosecond science and facilitated ultrafast spectroscopy in atoms, molecules, and solids. We explore high-harmonic generation in the solid state by means of nanostructured and ion-implanted semiconductors. We use wavelength-selective microscopic imaging to map enhanced harmonic emission and show that the generation medium and the driving field can be locally tailored in solids by modifying the chemical composition and morphology. This enables the control of high-harmonic technology within precisely engineered solid targets. We demonstrate customized high-harmonic wave fields with wavelengths down to 225 nanometers (ninth-harmonic order of 2-micrometer laser pulses) and present an integrated Fresnel zone plate target in silicon, which leads to diffraction-limited self-focusing of the generated harmonics down to 1-micrometer spot sizes.

Experimental Characterization of Nonlinear Harmonic Radiation from a Visible SASE FEL at Saturation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cornacchia, Massimo

2002-08-19

Nonlinear harmonic radiation was observed using the VISA SASE FEL at saturation. The gain lengths, spectra and energies of the three lowest SASE FEL modes were experimentally characterized. Both the measured harmonic gain lengths and center spectral wavelengths are shown to decrease with harmonic number, n, which is consistent with nonlinear harmonic theory. The measured energies for both second and third harmonics are about 1% of the fundamental; the strong second harmonic radiation ({approx} 1 {micro}J) observed is unique for low energy SASE FELs. These experimental results demonstrate for the first time the feasibility of using nonlinear harmonic SASE FELmore » radiation to produce coherent, femtosecond X-rays.« less

Fast epi-detected broadband multiplex CARS and SHG imaging of mouse skull cells

PubMed Central

Capitaine, Erwan; Moussa, Nawel Ould; Louot, Christophe; Bardet, Sylvia M.; Kano, Hideaki; Duponchel, Ludovic; Lévêque, Philippe; Couderc, Vincent; Leproux, Philippe

2017-01-01

We present a bimodal imaging system able to obtain epi-detected mutiplex coherent anti-Stokes Raman scattering (M-CARS) and second harmonic generation (SHG) signals coming from biological samples. We studied a fragment of mouse parietal bone and could detect broadband anti-Stokes and SHG responses originating from bone cells and collagen respectively. In addition we compared two post-processing methods to retrieve the imaginary part of the third-order nonlinear susceptibility related to the spontaneous Raman scattering. PMID:29359100

Comparing Yb-fiber and Ti:Sapphire lasers for depth resolved imaging of human skin (Conference Presentation)

NASA Astrophysics Data System (ADS)

Balu, Mihaela; Saytashev, Ilyas; Hou, Jue; Dantus, Marcos; Tromberg, Bruce J.

2016-02-01

We report on a direct comparison between Ti:Sapphire and Yb fiber lasers for depth-resolved label-free multimodal imaging of human skin. We found that the penetration depth achieved with the Yb laser was 80% greater than for the Ti:Sapphire. Third harmonic generation (THG) imaging with Yb laser excitation provides additional information about skin structure. Our results indicate the potential of fiber-based laser systems for moving into clinical use.

Harmonic generation and parametric decay in the ion cyclotron frequency range

DOE Office of Scientific and Technical Information (OSTI.GOV)

Skiff, F.N.; Wong, K.L.; Ono, M.

1984-06-01

Harmonic generation and parametric decay are examined in a toroidal ACT-I plasma using electrostatic plate antennas. The harmonic generation, which is consistent with sheath rectification, is sufficiently strong that the nonlinearly generated harmonic modes themselves decay parametrically. Resonant and nonresonant parametric decay of the second harmonic are observed and compared with uniform pump theory. Resonant decay of lower hybrid waves into lower hybrid waves and slow ion cyclotron waves is seen for the first time. Surprisingly, the decay processes are nonlinearly saturated, indicating absolute instability.

Painlevé IV Solutions from Hamiltonians with Equidistant Gapped Spectrum

NASA Astrophysics Data System (ADS)

Estrada-Delgado, M. I.; Fernández C, D. J.

2016-03-01

Supersymmetry transformations are applied to the harmonic oscillator for generating potentials Vk j whose spectra have a gap with respect to the initial one. The extremal states are found and, as the reduction theorem conditions are satisfied, ensuring that the system has third order ladder operators and it is connected with Painlevé IV (PIV) equation, then solutions to this equation can be generated. An alternative transformation is applied, by adding the levels needed to recover the spectrum of Vk j . The extremal states are found and, as the reduction theorem is met again, we get also solutions to the PIV equation which will be analysed.

Polar phase transitions in heteroepitaxial stabilized La0.5Y0.5AlO3 thin films

NASA Astrophysics Data System (ADS)

Liu, Shenghua; Zhang, Chunfeng; Zhu, Mengya; He, Qian; Chakhalian, Jak; Liu, Xiaoran; Borisevich, Albina; Wang, Xiaoyong; Xiao, Min

2017-10-01

We report on the fabrication of epitaxial La0.5Y0.5AlO3 ultrathin films on (001) LaAlO3 substrates. Structural characterizations by scanning transmission electron microscopy and x-ray diffraction confirm the high quality of the film with a - b + c - AlO6 octahedral tilt pattern. Unlike either of the nonpolar parent compound, LaAlO3 and YAlO3, second harmonic generation measurements on the thin films suggest a nonpolar-polar phase transition at T c near 500 K, and a polar-polar phase transition at T a near 160 K. By fitting the angular dependence of the second harmonic intensities, we further propose that the two polar structures can be assigned to the Pmc2 1 and Pmn2 1 space group, while the high temperature nonpolar structure belongs to the Pbnm space group.

Simultaneous negative refraction and focusing of fundamental frequency and second-harmonic fields by two-dimensional photonic crystals

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Jun; College of Physics and Electronic Engineering, Henan Normal University, 453007 Xinxiang, Henan; Zhang, Xiangdong, E-mail: zhangxd@bit.edu.cn

2015-09-28

Simultaneous negative refraction for both the fundamental frequency (FF) and second-harmonic (SH) fields in two-dimensional nonlinear photonic crystals have been found through both the physical analysis and exact numerical simulation. By combining such a property with the phase-matching condition and strong second-order susceptibility, we have designed a SH lens to realize focusing for both the FF and SH fields at the same time. Good-quality non-near field images for both FF and SH fields have been observed. The physical mechanism for such SH focusing phenomena has been disclosed, which is different from the backward SH generation as has been pointed outmore » in the previous investigations. In addition, the effect of absorption losses on the phenomena has also been discussed. Thus, potential applications of these phenomena to biphotonic microscopy technique are anticipated.« less

Coexistence of Weak Ferromagnetism and Ferroelectricity in the High Pressure LiNbO3-Type Phase of FeTiO3

NASA Astrophysics Data System (ADS)

Varga, T.; Kumar, A.; Vlahos, E.; Denev, S.; Park, M.; Hong, S.; Sanehira, T.; Wang, Y.; Fennie, C. J.; Streiffer, S. K.; Ke, X.; Schiffer, P.; Gopalan, V.; Mitchell, J. F.

2009-07-01

We report the magnetic and electrical characteristics of polycrystalline FeTiO3 synthesized at high pressure that is isostructural with acentric LiNbO3 (LBO). Piezoresponse force microscopy, optical second harmonic generation, and magnetometry demonstrate ferroelectricity at and below room temperature and weak ferromagnetism below ˜120K. These results validate symmetry-based criteria and first-principles calculations of the coexistence of ferroelectricity and weak ferromagnetism in a series of transition metal titanates crystallizing in the LBO structure.

A multiphoton objective design with incorporated beam splitter for enhanced fluorescence collection

PubMed Central

McMullen, Jesse D.; Zipfel, Warren R.

2010-01-01

We present a de novo design of an objective for use in multi-photon (MPM) and second harmonic generation (SHG) microscopy. This objective was designed to have a large field of view (FOV), while maintaining a moderate numerical aperture (NA) and relative straight forward construction. A dichroic beam splitter was incorporated within the objective itself allowing for an increase in the front aperture of the objective and corresponding enhancement of the solid angle of collected emission by an order of magnitude over existing designs. PMID:20389554

A multiphoton objective design with incorporated beam splitter for enhanced fluorescence collection.

PubMed

McMullen, Jesse D; Zipfel, Warren R

2010-03-15

We present a de novo design of an objective for use in multi-photon (MPM) and second harmonic generation (SHG) microscopy. This objective was designed to have a large field of view (FOV), while maintaining a moderate numerical aperture (NA) and relative straight forward construction. A dichroic beam splitter was incorporated within the objective itself allowing for an increase in the front aperture of the objective and corresponding enhancement of the solid angle of collected emission by an order of magnitude over existing designs.

Coexistence of weak ferromagnetism and ferroelectricity in the high pressure LiNbO3-type phase of FeTiO3.

PubMed

Varga, T; Kumar, A; Vlahos, E; Denev, S; Park, M; Hong, S; Sanehira, T; Wang, Y; Fennie, C J; Streiffer, S K; Ke, X; Schiffer, P; Gopalan, V; Mitchell, J F

2009-07-24

We report the magnetic and electrical characteristics of polycrystalline FeTiO_{3} synthesized at high pressure that is isostructural with acentric LiNbO_{3} (LBO). Piezoresponse force microscopy, optical second harmonic generation, and magnetometry demonstrate ferroelectricity at and below room temperature and weak ferromagnetism below approximately 120 K. These results validate symmetry-based criteria and first-principles calculations of the coexistence of ferroelectricity and weak ferromagnetism in a series of transition metal titanates crystallizing in the LBO structure.

Polarization retention in ultra-thin barium titanate films on Ge(001)

NASA Astrophysics Data System (ADS)

Cho, Yujin; Ponath, Patrick; Zheng, Lu; Hatanpaa, Benjamin; Lai, Keji; Demkov, Alexander A.; Downer, Michael C.

2018-04-01

We investigate polarization retention in 10 to 19 nm thick ferroelectric BaTiO3 (BTO) grown on Ge(001) by molecular beam epitaxy. The out-of-plane direction and reversibility of electric polarization were confirmed using piezoresponse force microscopy. After reverse-poling selected regions of the BTO films to a value P with a biased atomic-force microscope tip, we monitored relaxation of their net polarization for as long as several weeks using optical second-harmonic generation microscopy. All films retained reversed polarization throughout the observation period. 10 nm-thick BTO films relaxed monotonically to a saturation value of 0.9 P after 27 days and 19 nm films to 0.75 P after 24 h. Polarization dynamics are discussed in the context of a 1D polarization relaxation/kinetics model.

Characterizing lamina propria of human gastric mucosa by multiphoton microscopy

NASA Astrophysics Data System (ADS)

Liu, Y. C.; Yang, H. Q.; Chen, G.; Zhuo, S. M.; Chen, J. X.; Yan, J.

2011-01-01

Lamina propria (LP) of gastric mucosa plays an important role in progression of gastric cancer because of the site at where inflammatory reactions occur. Multiphoton imaging has been recently employed for microscopic examination of intact tissue. In this paper, using multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG), high resolution multiphoton microscopic images of lamina propria (LP) are obtained in normal human gastric mucosa at excitation wavelength λex = 800 nm. The main source of tissue TPEF originated from the cells of gastric glands, and loose connective tissue, collagen, produced SHG signals. Our results demonstrated that MPM can be effective for characterizing the microstructure of LP in human gastric mucosa. The findings will be helpful for diagnosing and staging early gastric cancer in the clinics.

In vivo three-dimensional optical coherence tomography and multiphoton microscopy in a mouse model of ovarian neoplasia

NASA Astrophysics Data System (ADS)

Watson, Jennifer M.; Marion, Samuel L.; Rice, Photini Faith; Bentley, David L.; Besselsen, David; Utzinger, Urs; Hoyer, Patricia B.; Barton, Jennifer K.

2013-03-01

Our goal is to use optical coherence tomography (OCT) and multiphoton microscopy (MPM) to detect early tumor development in a mouse model of ovarian neoplasia. We hope to use information regarding early tumor development to create a diagnostic test for high-risk patients. In this study we collect in vivo images using OCT, second harmonic generation and two-photon excited fluorescence from non-vinylcyclohexene diepoxide (VCD)-dosed and VCD-dosed mice. VCD causes follicular apoptosis (simulating menopause) and leads to tumor development. Using OCT and MPM we visualized the ovarian microstructure and were able to see differences between non-VCD-dosed and VCD-dosed animals. This leads us to believe that OCT and MPM may be useful for detecting changes due to early tumor development.

Evaluation of area-based collagen scoring by nonlinear microscopy in chronic hepatitis C-induced liver fibrosis

PubMed Central

Sevrain, David; Dubreuil, Matthieu; Dolman, Grace Elizabeth; Zaitoun, Abed; Irving, William; Guha, Indra Neil; Odin, Christophe; Le Grand, Yann

2015-01-01

In this paper we analyze a fibrosis scoring method based on measurement of the fibrillar collagen area from second harmonic generation (SHG) microscopy images of unstained histological slices from human liver biopsies. The study is conducted on a cohort of one hundred chronic hepatitis C patients with intermediate to strong Metavir and Ishak stages of liver fibrosis. We highlight a key parameter of our scoring method to discriminate between high and low fibrosis stages. Moreover, according to the intensity histograms of the SHG images and simple mathematical arguments, we show that our area-based method is equivalent to an intensity-based method, despite saturation of the images. Finally we propose an improvement of our scoring method using very simple image processing tools. PMID:25909005

Evaluation of area-based collagen scoring by nonlinear microscopy in chronic hepatitis C-induced liver fibrosis.

PubMed

Sevrain, David; Dubreuil, Matthieu; Dolman, Grace Elizabeth; Zaitoun, Abed; Irving, William; Guha, Indra Neil; Odin, Christophe; Le Grand, Yann

2015-04-01

In this paper we analyze a fibrosis scoring method based on measurement of the fibrillar collagen area from second harmonic generation (SHG) microscopy images of unstained histological slices from human liver biopsies. The study is conducted on a cohort of one hundred chronic hepatitis C patients with intermediate to strong Metavir and Ishak stages of liver fibrosis. We highlight a key parameter of our scoring method to discriminate between high and low fibrosis stages. Moreover, according to the intensity histograms of the SHG images and simple mathematical arguments, we show that our area-based method is equivalent to an intensity-based method, despite saturation of the images. Finally we propose an improvement of our scoring method using very simple image processing tools.

Dynamical origin of near- and below-threshold harmonic generation of Cs in an intense mid-infrared laser field.

PubMed

Li, Peng-Cheng; Sheu, Yae-Lin; Laughlin, Cecil; Chu, Shih-I

2015-05-20

Near- and below-threshold harmonic generation provides a potential approach to generate vacuum-ultraviolet frequency comb. However, the dynamical origin of in these lower harmonics is less understood and largely unexplored. Here we perform an ab initio quantum study of the near- and below-threshold harmonic generation of caesium (Cs) atoms in an intense 3,600-nm mid-infrared laser field. Combining with a synchrosqueezing transform of the quantum time-frequency spectrum and an extended semiclassical analysis, the roles of multiphoton and multiple rescattering trajectories on the near- and below-threshold harmonic generation processes are clarified. We find that the multiphoton-dominated trajectories only involve the electrons scattered off the higher part of the combined atom-field potential followed by the absorption of many photons in near- and below-threshold regime. Furthermore, only the near-resonant below-threshold harmonic is exclusive to exhibit phase locked features. Our results shed light on the dynamic origin of the near- and below-threshold harmonic generation.

Non-phase-matched enhancement of second-harmonic generation in multilayer nonlinear structures with internal reflections.

PubMed

Centini, Marco; D'Aguanno, Giuseppe; Sciscione, Letizia; Sibilia, Concita; Bertolotti, Mario; Scalora, Michael; Bloemer, Mark J

2004-08-15

Traditional notions of second-harmonic generation rely on phase matching or quasi phase matching to achieve good conversion efficiencies. We present an entirely new concept for efficient second-harmonic generation that is based on the interference of counterpropagating waves in multilayer structures. Conversion efficiencies are an order of magnitude larger than with phase-matched second-harmonic generation in similar multilayer structures.

Weakly nonlinear incompressible Rayleigh-Taylor instability in spherical and planar geometries

NASA Astrophysics Data System (ADS)

Zhang, J.; Wang, L. F.; Ye, W. H.; Guo, H. Y.; Wu, J. F.; Ding, Y. K.; Zhang, W. Y.; He, X. T.

2018-02-01

The relationship between the weakly nonlinear (WN) solutions of the Rayleigh-Taylor instability in spherical geometry [Zhang et al., Phys. Plasmas 24, 062703 (2017)] and those in planar geometry [Wang et al., Phys. Plasmas 19, 112706 (2012)] is analyzed. In the high-mode perturbation limit ( Pn(cos θ), n ≫1 ), it is found that at the equator, the contributions of mode P2 n along with its neighboring modes, mode P3 n along with its neighboring modes, and mode Pn at the third order along with its neighboring modes are equal to those of the second harmonic, the third harmonic, and the third-order feedback to the fundamental mode, respectively, in the planar case with a perturbation of the same wave vector and amplitude as those at the equator. The trends of WN results in spherical geometry towards the corresponding planar counterparts are found, and the convergence behaviors of the neighboring modes of Pn, P2 n , and P3 n are analyzed. Moreover, the spectra generated from the high-mode perturbations in the WN regime are provided. For low-mode perturbations, it is found that the fundamental modes saturate at larger amplitudes than the planar result. The geometry effect makes the bubbles at or near the equator grow faster than the bubbles in planar geometry in the WN regime.

Symmetry properties of second harmonics generated by antisymmetric Lamb waves

NASA Astrophysics Data System (ADS)

Zhu, Wujun; Xiang, Yanxun; Liu, Chang-Jun; Deng, Mingxi; Xuan, Fu-Zhen

2018-03-01

Symmetry properties of second harmonics generated by antisymmetric primary Lamb waves are systematically studied in this work. In theory, the acoustic field of second harmonic Lamb waves is obtained by using the perturbation approximation and normal modal method, and the energy flux transfer from the primary Lamb waves to second harmonics is mainly explored. Symmetry analyses indicate that either the symmetric or antisymmetric Lamb waves can merely generate the symmetric second harmonics. Finite element simulations are performed on the nonlinear Lamb wave propagation of the antisymmetric A0 mode in the low frequency region. The signals of the second harmonics and the symmetric second harmonic s0 mode are found to be exactly equivalent in the time domain. The relative acoustic nonlinearity parameter A2/A12 oscillates with the propagation distance, and the oscillation amplitude and spatial period are well consistent with the theoretical prediction of the A0-s0 mode pair, which means that only the second harmonic s0 mode is generated by the antisymmetric primary A0 mode. Experiments are further conducted to examine the cumulative generation of symmetric second harmonics for the antisymmetric-symmetric mode pair A3-s6. Results show that A2/A12 increases linearly with the propagation distance, which means that the symmetric second harmonic s6 mode is generated cumulatively by the antisymmetric primary A3 mode. The present investigation systematically corroborates the proposed theory that only symmetric second harmonics can be generated accompanying the propagation of antisymmetric primary Lamb waves in a plate.

Surface diffusion of In on Ge(111) studied by optical second harmonic microscopy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Suni, I.I.; Seebauer, E.G.

Surface diffusion of In on Ge(111) has been measured by optical second harmonic microscopy. This technique employs surface second harmonic generation to directly image submonolayer surface concentration profiles. The coverage dependence of the diffusivity [ital D] can then be obtained from a Boltzmann--Matano analysis. In the coverage range 0.1[lt][theta][lt]0.48, the activation energy [ital E][sub diff] decreased with increasing coverage, ranging from 31 kcal/mol at [theta]=0.1 to 23 kcal/mol at [theta]=0.48. Over the same coverage range, the pre-exponential factor [ital D][sub 0] decreased from 5[times]10[sup 2] to 1[times]10[sup [minus]1] cm[sup 2]/s. This gradual change reflects a change in diffusion mechanism arisingmore » from the disordered nature of the Ge(111) surface. At low coverages, In adatoms sink into the top layer of Ge, and diffusion is dominated by thermal formation of adatom-vacancy pairs. At high coverages, diffusion occurs by normal site-to-site hopping. The gradual change in diffusion parameters with coverage was interrupted by an apparent phase transition at [theta]=0.16. At this point, both [ital E][sub diff] and [ital D][sub 0] peaked sharply at 41 kcal/mol and 6[times]10[sup 5] cm[sup 2]/s, respectively. The desorption energy [ital E][sub des] was measured by temperature programmed desorption. [ital E][sub des] decreased from 60 kcal/mol at submonolayer coverages to 55 kcal/mol at multilayer coverages.« less

Tunneling ionization and harmonic generation in two-color fields

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kondo, K.; Kobayashi, Y.; Sagisaka, A.

1996-02-01

Tunneling ionization and harmonic generation in two-color fields were studied with a fundamental beam ({omega}) and its harmonics (2{omega},3{omega}), which were generated by a 100-fs Ti:sapphire laser. Ion yields of atoms and molecules were successfully controlled by means of a change in the relative phase between {omega} and 3{omega} pulses. Two-color interference was clearly observed in photoelectron spectra and harmonic spectra. In the {omega}{endash}2{omega} field even-order harmonics were observed in which the intensity was almost equal to that of the odd harmonics because of an asymmetric optical field. These results were compared with the quasi-static model for ionization and withmore » the quantum theory for harmonic generation. {copyright} {ital 1996 Optical Society of America.}« less

Non-linear dielectric spectroscopy of microbiological suspensions

PubMed Central

Treo, Ernesto F; Felice, Carmelo J

2009-01-01

Background Non-linear dielectric spectroscopy (NLDS) of microorganism was characterized by the generation of harmonics in the polarization current when a microorganism suspension was exposed to a sinusoidal electric field. The biological nonlinear response initially described was not well verified by other authors and the results were susceptible to ambiguous interpretation. In this paper NLDS was performed to yeast suspension in tripolar and tetrapolar configuration with a recently developed analyzer. Methods Tripolar analysis was carried out by applying sinusoidal voltages up to 1 V at the electrode interface. Tetrapolar analysis was carried on with sinusoidal field strengths from 0.1 V cm-1 to 70 V cm-1. Both analyses were performed within a frequency range from 1 Hz through 100 Hz. The harmonic amplitudes were Fourier-analyzed and expressed in dB. The third harmonic, as reported previously, was investigated. Statistical analysis (ANOVA) was used to test the effect of inhibitor an activator of the plasma membrane enzyme in the measured response. Results No significant non-linearities were observed in tetrapolar analysis, and no observable changes occurred when inhibitor and activator were added to the suspension. Statistical analysis confirmed these results. When a pure sinus voltage was applied to an electrode-yeast suspension interface, variations higher than 25 dB for the 3rd harmonic were observed. Variation higher than 20 dB in the 3rd harmonics has also been found when adding an inhibitor or activator of the membrane-bounded enzymes. These variations did not occur when the suspension was boiled. Discussion The lack of result in tetrapolar cells suggest that there is no, if any, harmonic generation in microbiological bulk suspension. The non-linear response observed was originated in the electrode-electrolyte interface. The frequency and voltage windows observed in previous tetrapolar analysis were repeated in the tripolar measurements, but maximum were not observed at the same values. Conclusion Contrary to previous assertions, no repeatable dielectric non-linearity was exhibited in the bulk suspensions tested under the field and frequency condition reported with this recently designed analyzer. Indeed, interface related harmonics were observed and monitored during biochemical stimuli. The changes were coherent with the expected biological response. PMID:19772595

Amplitude-dependent internal friction, hysteretic nonlinearity, and nonlinear oscillations in a magnesite resonator.

PubMed

Nazarov, V E; Kolpakov, A B; Radostin, A V

2012-07-01

The results of experimental and theoretical studies of low-frequency nonlinear acoustics phenomena (amplitude-dependent loss, resonance frequency shifts, and a generation of second and third harmonics) in a magnesite rod resonator are presented. Acceleration and velocity oscillograms of vibrations of the free boundary of the resonator caused by harmonic excitations were measured and analyzed. A theoretical description of the observed amplitude dependences was carried out within the framework of the phenomenological state equations that contain either of the two types of hysteretic nonlinearity (elastic and inelastic). The type of hysteresis and parameters of acoustic nonlinearity of magnesite were established from comparing the experimental measurements with the theoretical dependences. The values of the parameters were anomalously high even when compared to those of other strongly nonlinear polycrystalline materials such as granite, marble, limestone, sandstone, etc.

Generations of even-order harmonics from vibrating H2+ and T2+ in the rising and falling parts of the laser field

NASA Astrophysics Data System (ADS)

Feng, Liqiang; Kapteyn, Henry J.; Feng, April Y.

2018-04-01

The generations of the even-order harmonics from H2+ and one of its isotope T2+ have been theoretically investigated beyond the Born-Oppenheimer approximation. Normally, the high-order harmonic generation (HHG) only contains odd-order harmonics for the orbital symmetry along the direction of laser polarization. Here, we showed that due to asymmetric harmonic emission (asymmetric half-wave profile), the even-order harmonics can be generated in the rising and the falling part of the laser field. In detail, in the lower initial vibrational state, the even-order harmonics main come from the falling part of the laser field; while as the initial vibrational state increases, the identified even-order harmonics in the falling part of the laser field are decreased; while some other even-order harmonics coming from the rising part of the laser field can be produced. The interesting phenomena have been proved through studying the spatial distributions and the time profiles of the HHG.

Power MOSFET Linearizer of a High-Voltage Power Amplifier for High-Frequency Pulse-Echo Instrumentation.

PubMed

Choi, Hojong; Woo, Park Chul; Yeom, Jung-Yeol; Yoon, Changhan

2017-04-04

A power MOSFET linearizer is proposed for a high-voltage power amplifier (HVPA) used in high-frequency pulse-echo instrumentation. The power MOSFET linearizer is composed of a DC bias-controlled series power MOSFET shunt with parallel inductors and capacitors. The proposed scheme is designed to improve the gain deviation characteristics of the HVPA at higher input powers. By controlling the MOSFET bias voltage in the linearizer, the gain reduction into the HVPA was compensated, thereby reducing the echo harmonic distortion components generated by the ultrasonic transducers. In order to verify the performance improvement of the HVPA implementing the power MOSFET linearizer, we measured and found that the gain deviation of the power MOSFET linearizer integrated with HVPA under 10 V DC bias voltage was reduced (-1.8 and -0.96 dB, respectively) compared to that of the HVPA without the power MOSFET linearizer (-2.95 and -3.0 dB, respectively) when 70 and 80 MHz, three-cycle, and 26 dB m input pulse waveforms are applied, respectively. The input 1-dB compression point (an index of linearity) of the HVPA with power MOSFET linearizer (24.17 and 26.19 dB m at 70 and 80 MHz, respectively) at 10 V DC bias voltage was increased compared to that of HVPA without the power MOSFET linearizer (22.03 and 22.13 dB m at 70 and 80 MHz, respectively). To further verify the reduction of the echo harmonic distortion components generated by the ultrasonic transducers, the pulse-echo responses in the pulse-echo instrumentation were compared when using HVPA with and without the power MOSFET linearizer. When three-cycle 26 dB m input power was applied, the second, third, fourth, and fifth harmonic distortion components of a 75 MHz transducer driven by the HVPA with power MOSFET linearizer (-48.34, -44.21, -48.34, and -46.56 dB, respectively) were lower than that of the HVPA without the power MOSFET linearizer (-45.61, -41.57, -45.01, and -45.51 dB, respectively). When five-cycle 20 dB m input power was applied, the second, third, fourth, and fifth harmonic distortions of the HVPA with the power MOSFET linearizer (-41.54, -41.80, -48.86, and -46.27 dB, respectively) were also lower than that of the HVPA without the power MOSFET linearizer (-25.85, -43.56, -49.04, and -49.24 dB, respectively). Therefore, we conclude that the power MOSFET linearizer could reduce gain deviation of the HVPA, thus reducing the echo signal harmonic distortions generated by the high-frequency ultrasonic transducers in pulse-echo instrumentation.

Power MOSFET Linearizer of a High-Voltage Power Amplifier for High-Frequency Pulse-Echo Instrumentation

PubMed Central

Choi, Hojong; Woo, Park Chul; Yeom, Jung-Yeol; Yoon, Changhan

2017-01-01

A power MOSFET linearizer is proposed for a high-voltage power amplifier (HVPA) used in high-frequency pulse-echo instrumentation. The power MOSFET linearizer is composed of a DC bias-controlled series power MOSFET shunt with parallel inductors and capacitors. The proposed scheme is designed to improve the gain deviation characteristics of the HVPA at higher input powers. By controlling the MOSFET bias voltage in the linearizer, the gain reduction into the HVPA was compensated, thereby reducing the echo harmonic distortion components generated by the ultrasonic transducers. In order to verify the performance improvement of the HVPA implementing the power MOSFET linearizer, we measured and found that the gain deviation of the power MOSFET linearizer integrated with HVPA under 10 V DC bias voltage was reduced (−1.8 and −0.96 dB, respectively) compared to that of the HVPA without the power MOSFET linearizer (−2.95 and −3.0 dB, respectively) when 70 and 80 MHz, three-cycle, and 26 dBm input pulse waveforms are applied, respectively. The input 1-dB compression point (an index of linearity) of the HVPA with power MOSFET linearizer (24.17 and 26.19 dBm at 70 and 80 MHz, respectively) at 10 V DC bias voltage was increased compared to that of HVPA without the power MOSFET linearizer (22.03 and 22.13 dBm at 70 and 80 MHz, respectively). To further verify the reduction of the echo harmonic distortion components generated by the ultrasonic transducers, the pulse-echo responses in the pulse-echo instrumentation were compared when using HVPA with and without the power MOSFET linearizer. When three-cycle 26 dBm input power was applied, the second, third, fourth, and fifth harmonic distortion components of a 75 MHz transducer driven by the HVPA with power MOSFET linearizer (−48.34, −44.21, −48.34, and −46.56 dB, respectively) were lower than that of the HVPA without the power MOSFET linearizer (−45.61, −41.57, −45.01, and −45.51 dB, respectively). When five-cycle 20 dBm input power was applied, the second, third, fourth, and fifth harmonic distortions of the HVPA with the power MOSFET linearizer (−41.54, −41.80, −48.86, and −46.27 dB, respectively) were also lower than that of the HVPA without the power MOSFET linearizer (−25.85, −43.56, −49.04, and −49.24 dB, respectively). Therefore, we conclude that the power MOSFET linearizer could reduce gain deviation of the HVPA, thus reducing the echo signal harmonic distortions generated by the high-frequency ultrasonic transducers in pulse-echo instrumentation. PMID:28375165

High-harmonic generation in amorphous solids

DOE Office of Scientific and Technical Information (OSTI.GOV)

You, Yong Sing; Yin, Yanchun; Wu, Yi

High-harmonic generation in isolated atoms and molecules has been widely utilized in extreme ultraviolet photonics and attosecond pulse metrology. Recently, high-harmonic generation has been observed in solids, which could lead to important applications such as all-optical methods to image valance charge density and reconstruct electronic band structures, as well as compact extreme ultraviolet light sources. So far these studies are confined to crystalline solids; therefore, decoupling the respective roles of long-range periodicity and high density has been challenging. Here we report the observation of high-harmonic generation from amorphous fused silica. We also decouple the role of long-range periodicity by comparingmore » harmonics generated from fused silica and crystalline quartz, which contain the same atomic constituents but differ in long-range periodicity. These results advance current understanding of the strong-field processes leading to high-harmonic generation in solids with implications for the development of robust and compact extreme ultraviolet light sources.« less

High-harmonic generation in amorphous solids

DOE PAGES

You, Yong Sing; Yin, Yanchun; Wu, Yi; ...

2017-09-28

High-harmonic generation in isolated atoms and molecules has been widely utilized in extreme ultraviolet photonics and attosecond pulse metrology. Recently, high-harmonic generation has been observed in solids, which could lead to important applications such as all-optical methods to image valance charge density and reconstruct electronic band structures, as well as compact extreme ultraviolet light sources. So far these studies are confined to crystalline solids; therefore, decoupling the respective roles of long-range periodicity and high density has been challenging. Here we report the observation of high-harmonic generation from amorphous fused silica. We also decouple the role of long-range periodicity by comparingmore » harmonics generated from fused silica and crystalline quartz, which contain the same atomic constituents but differ in long-range periodicity. These results advance current understanding of the strong-field processes leading to high-harmonic generation in solids with implications for the development of robust and compact extreme ultraviolet light sources.« less

Carcinogenic damage to deoxyribonucleic acid is induced by near-infrared laser pulses in multiphoton microscopy via combination of two- and three-photon absorption

NASA Astrophysics Data System (ADS)

Nadiarnykh, Oleg; Thomas, Giju; Van Voskuilen, Johan; Sterenborg, Henricus J. C. M.; Gerritsen, Hans C.

2012-11-01

Nonlinear optical imaging modalities (multiphoton excited fluorescence, second and third harmonic generation) applied in vivo are increasingly promising for clinical diagnostics and the monitoring of cancer and other disorders, as they can probe tissue with high diffraction-limited resolution at near-infrared (IR) wavelengths. However, high peak intensity of femtosecond laser pulses required for two-photon processes causes formation of cyclobutane-pyrimidine-dimers (CPDs) in cellular deoxyribonucleic acid (DNA) similar to damage from exposure to solar ultraviolet (UV) light. Inaccurate repair of subsequent mutations increases the risk of carcinogenesis. In this study, we investigate CPD damage that results in Chinese hamster ovary cells in vitro from imaging them with two-photon excited autofluorescence. The CPD levels are quantified by immunofluorescent staining. We further evaluate the extent of CPD damage with respect to varied wavelength, pulse width at focal plane, and pixel dwell time as compared with more pronounced damage from UV sources. While CPD damage has been expected to result from three-photon absorption, our results reveal that CPDs are induced by competing two- and three-photon absorption processes, where the former accesses UVA absorption band. This finding is independently confirmed by nonlinear dependencies of damage on laser power, wavelength, and pulse width.

A study of medical device regulation management model in Asia.

PubMed

Wu, Yi-Hui; Li, Fong-An; Fan, Yin-Ting; Tu, Pei-Weng

2016-06-01

With the aging of the post-war baby boomer generation, the increasing demands for healthcare are driving the growth of medical industry and development of new products in order to meet the immense needs from the aging population. However, medical devices are designed to maintain the health and safety of people, therefore, medical devices are undergoing rigorous management by competent health authorities in all countries. In recent years, Asian countries have been reforming their regulations and standards for medical devices with substantial changes. The study is a summary of the framework of medical device regulations in Asian countries, including Asian Harmonization Working Party (AHWP), Japan, China, Taiwan, South Korea, India and Singapore. Expert commentary: Asian countries are constantly reforming their medical device regulations. The emergence of brand-new technology and quality management issues arose by global manufacturing have imposed difficulties in harmonizing and reaching consensus between countries. The third-party conformity assessment system for medical devices can reduce the costs for competent health authorities and shorten the review time, which could facilitate the feasibility of harmonization of medical device regulations.

Second harmonic generation at fatigue cracks by low-frequency Lamb waves: Experimental and numerical studies

NASA Astrophysics Data System (ADS)

Yang, Yi; Ng, Ching-Tai; Kotousov, Andrei; Sohn, Hoon; Lim, Hyung Jin

2018-01-01

This paper presents experimental and theoretical analyses of the second harmonic generation due to non-linear interaction of Lamb waves with a fatigue crack. Three-dimensional (3D) finite element (FE) simulations and experimental studies are carried out to provide physical insight into the mechanism of second harmonic generation. The results demonstrate that the 3D FE simulations can provide a reasonable prediction on the second harmonic generated due to the contact nonlinearity at the fatigue crack. The effect of the wave modes on the second harmonic generation is also investigated in detail. It is found that the magnitude of the second harmonic induced by the interaction of the fundamental symmetric mode (S0) of Lamb wave with the fatigue crack is much higher than that by the fundamental anti-symmetric mode (A0) of Lamb wave. In addition, a series of parametric studies using 3D FE simulations are conducted to investigate the effect of the fatigue crack length to incident wave wavelength ratio, and the influence of the excitation frequency on the second harmonic generation. The outcomes show that the magnitude and directivity pattern of the generated second harmonic depend on the fatigue crack length to incident wave wavelength ratio as well as the ratio of S0 to A0 incident Lamb wave amplitude. In summary, the findings of this study can further advance the use of second harmonic generation in damage detection.

Nonlinear optical anisotropy and molecular orientational distribution in poly(p-phenylene benzobisthiazole) Langmuir-Blodgett films

NASA Astrophysics Data System (ADS)

Wang, Liming; Wada, Tatsuo; Yuba, Tomoyuki; Kakimoto, Masaaki; Imai, Yoshio; Sasabe, Hiroyuki

1996-06-01

The orientational distribution and packing of polymer chains were investigated in poly(p-phenylene benzobisthiazole) (PBT) Langmuir-Blodgett (LB) films by nonresonant third-harmonic generation measurement at a wavelength of 1907 nm. The tensor components of the third-harmonic susceptibility on the PBT LB film with a surface pressure of 50 mN/m were determined to be χ(3)XXXX=(16.6±2.5)×10-12 and χ(3)YYYY=(2.0±0.3)×10-12. The large nonlinear optical anisotropy can be explained as a result of highly oriented packing of the polymer chains induced by a flow orientation. A Gaussian distribution function with a standard deviation of σ=0.40 gives a practical description of the orientational distribution of PBT polymer chains. A maximum χ(3) value of (26.8±4.4)×10-12 esu is predicted assuming a perfect alignment of polymer chains. The χ(3)XXXX value increased by factor of 2 with the surface pressure from 30 to 50 mN/m mainly due to the packing density of the polymer chains, while the orientational degree did not change.

Application of mid-infrared pulses for quasi-phase-matching of high-order harmonics in silver plasma.

PubMed

Ganeev, Rashid A; Husakou, Anton; Suzuki, Masayuki; Kuroda, Hiroto

2016-02-22

We demonstrate the quasi-phase-matching of a group of harmonics generated in Ag multi-jet plasma using tunable pulses in the region of 1160 - 1540 nm and their second harmonic emission. The numerical treatment of this effect includes microscopic description of the harmonic generation, propagation of the pump pulse, and the propagation of the generated harmonics. We obtained more than 30-fold growth of harmonics at the conditions of quasi-phase-matching in the region of 35 nm using eight-jet plasma compared with the case of imperforated plasma.

Resonant second harmonic generation in a gallium nitride two-dimensional photonic crystal on silicon

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zeng, Y.; Roland, I.; Checoury, X.

We demonstrate second harmonic generation in a gallium nitride photonic crystal cavity embedded in a two-dimensional free-standing photonic crystal platform on silicon. The photonic crystal nanocavity is optically pumped with a continuous-wave laser at telecom wavelengths in the transparency window of the nitride material. The harmonic generation is evidenced by the spectral range of the emitted signal, the quadratic power dependence vs. input power, and the spectral dependence of second harmonic signal. The harmonic emission pattern is correlated to the harmonic polarization generated by the second-order nonlinear susceptibilities χ{sub zxx}{sup (2)}, χ{sub zyy}{sup (2)} and the electric fields of the fundamentalmore » cavity mode.« less

Phase Coupling Between Spectral Components of Collapsing Langmuir Solitons in Solar Type III Radio Bursts

NASA Technical Reports Server (NTRS)

Thejappa, G.; MacDowall, R. J.; Bergamo, M.

2012-01-01

We present the high time resolution observations of one of the Langmuir wave packets obtained in the source region of a solar type III radio burst. This wave packet satisfies the threshold condition of the supersonic modulational instability, as well as the criterion of a collapsing Langmuir soliton, i.e., the spatial scale derived from its peak intensity is less than that derived from its short time scale. The spectrum of t his wave packet contains an intense spectral peak at local electron plasma frequency, f(sub pe) and relatively weaker peaks at 2f(sub pe) and 3f(sub pe). We apply the wavelet based bispectral analysis technique on this wave packet and compute the bicoherence between its spectral components. It is found that the bicoherence exhibits two peaks at (approximately f(sub pe), approximately f(sub pe)) and (approximately f(sub pe) approximately 2f(sub pe)), which strongly suggest that the spectral peak at 2f(sub pe) probably corresponds to the second harmonic radio emission, generated as a result of the merging of antiparallel propagating Langmuir waves trapped in the collapsing Langmuir soliton, and, the spectral peak at 3f(sub pe) probably corresponds to the third harmonic radio emission, generated as a result of merging of a trapped Langmuir wave and a second harmonic electromagnetic wave.

Fundamental and subharmonic excitation for an oscillator with several tunneling diodes in series

NASA Technical Reports Server (NTRS)

Boric-Lubecke, Olga; Pan, Dee-Son; Itoh, Tatsuo

1995-01-01

Connecting several tunneling diodes in series shows promise as a method for increasing the output power of these devices as millimeter-wave oscillators. However, due to the negative differential resistance (NDR) region in the dc I-V curve of a single tunneling diode, a circuit using several devices connected in series, and biased simultaneously in the NDR region, is dc unstable. Because of this instability, an oscillator with several tunneling diodes in series has a demanding excitation condition. Excitation using an externally applied RF signal is one approach to solving this problem. This is experimentally demonstrated using an RF source, both with frequency close to as well as with frequency considerably lower than the oscillation frequency. Excitation by an RF (radio frequency) source with a frequency as low as one sixth of the oscillation frequency was demonstrated in a proof-of-principle experiment at 2 GHz, for an oscillator with two tunnel diodes connected in series. Strong harmonics of the oscillation signal were generated as a result of the highly nonlinear dc I-V curve of the tunnel diode and a large signal oscillator design. Third harmonic output power comparable to that of the fundamental was observed in one oscillator circuit. If submillimeter wave resonant-tunneling diodes (RTD's) are used instead of tunnel diodes, this harmonic output may be useful for generating signals at frequencies well into the terahertz range.

A look-up-table digital predistortion technique for high-voltage power amplifiers in ultrasonic applications.

PubMed

Gao, Zheng; Gui, Ping

2012-07-01

In this paper, we present a digital predistortion technique to improve the linearity and power efficiency of a high-voltage class-AB power amplifier (PA) for ultrasound transmitters. The system is composed of a digital-to-analog converter (DAC), an analog-to-digital converter (ADC), and a field-programmable gate array (FPGA) in which the digital predistortion (DPD) algorithm is implemented. The DPD algorithm updates the error, which is the difference between the ideal signal and the attenuated distorted output signal, in the look-up table (LUT) memory during each cycle of a sinusoidal signal using the least-mean-square (LMS) algorithm. On the next signal cycle, the error data are used to equalize the signal with negative harmonic components to cancel the amplifier's nonlinear response. The algorithm also includes a linear interpolation method applied to the windowed sinusoidal signals for the B-mode and Doppler modes. The measurement test bench uses an arbitrary function generator as the DAC to generate the input signal, an oscilloscope as the ADC to capture the output waveform, and software to implement the DPD algorithm. The measurement results show that the proposed system is able to reduce the second-order harmonic distortion (HD2) by 20 dB and the third-order harmonic distortion (HD3) by 14.5 dB, while at the same time improving the power efficiency by 18%.

Quantitative characterization of collagen in the fibrotic capsule surrounding implanted polymeric microparticles through second harmonic generation imaging

DOE PAGES

Akilbekova, Dana; Bratlie, Kaitlin M.; Abraham, Thomas

2015-06-30

The collagenous capsule formed around an implant will ultimately determine the nature of its in vivo fate. To provide a better understanding of how surface modifications can alter the collagen orientation and composition in the fibrotic capsule, we used second harmonic generation (SHG) microscopy to evaluate collagen organization and structure generated in mice subcutaneously injected with chemically functionalized polystyrene particles. SHG is sensitive to the orientation of a molecule, making it a powerful tool for measuring the alignment of collagen fibers. Additionally, SHG arises from the second order susceptibility of the interrogated molecule in response to the electric field. Variationmore » in these tensor components distinguishes different molecular sources of SHG, providing collagen type specificity. Here, we demonstrated the ability of SHG to differentiate collagen type I and type III quantitatively and used this method to examine fibrous capsules of implanted polystyrene particles. Data presented in this work shows a wide range of collagen fiber orientations and collagen compositions in response to surface functionalized polystyrene particles. Dimethylamino functionalized particles were able to form a thin collagenous matrix resembling healthy skin. These findings have the potential to improve the fundamental understanding of how material properties influence collagen organization and composition quantitatively.« less

Quantitative Characterization of Collagen in the Fibrotic Capsule Surrounding Implanted Polymeric Microparticles through Second Harmonic Generation Imaging.

PubMed

Akilbekova, Dana; Bratlie, Kaitlin M

2015-01-01

The collagenous capsule formed around an implant will ultimately determine the nature of its in vivo fate. To provide a better understanding of how surface modifications can alter the collagen orientation and composition in the fibrotic capsule, we used second harmonic generation (SHG) microscopy to evaluate collagen organization and structure generated in mice subcutaneously injected with chemically functionalized polystyrene particles. SHG is sensitive to the orientation of a molecule, making it a powerful tool for measuring the alignment of collagen fibers. Additionally, SHG arises from the second order susceptibility of the interrogated molecule in response to the electric field. Variation in these tensor components distinguishes different molecular sources of SHG, providing collagen type specificity. Here, we demonstrated the ability of SHG to differentiate collagen type I and type III quantitatively and used this method to examine fibrous capsules of implanted polystyrene particles. Data presented in this work shows a wide range of collagen fiber orientations and collagen compositions in response to surface functionalized polystyrene particles. Dimethylamino functionalized particles were able to form a thin collagenous matrix resembling healthy skin. These findings have the potential to improve the fundamental understanding of how material properties influence collagen organization and composition quantitatively.

Quantitative Characterization of Collagen in the Fibrotic Capsule Surrounding Implanted Polymeric Microparticles through Second Harmonic Generation Imaging

PubMed Central

Akilbekova, Dana; Bratlie, Kaitlin M.

2015-01-01

The collagenous capsule formed around an implant will ultimately determine the nature of its in vivo fate. To provide a better understanding of how surface modifications can alter the collagen orientation and composition in the fibrotic capsule, we used second harmonic generation (SHG) microscopy to evaluate collagen organization and structure generated in mice subcutaneously injected with chemically functionalized polystyrene particles. SHG is sensitive to the orientation of a molecule, making it a powerful tool for measuring the alignment of collagen fibers. Additionally, SHG arises from the second order susceptibility of the interrogated molecule in response to the electric field. Variation in these tensor components distinguishes different molecular sources of SHG, providing collagen type specificity. Here, we demonstrated the ability of SHG to differentiate collagen type I and type III quantitatively and used this method to examine fibrous capsules of implanted polystyrene particles. Data presented in this work shows a wide range of collagen fiber orientations and collagen compositions in response to surface functionalized polystyrene particles. Dimethylamino functionalized particles were able to form a thin collagenous matrix resembling healthy skin. These findings have the potential to improve the fundamental understanding of how material properties influence collagen organization and composition quantitatively. PMID:26125551

Quantitative characterization of collagen in the fibrotic capsule surrounding implanted polymeric microparticles through second harmonic generation imaging

DOE Office of Scientific and Technical Information (OSTI.GOV)

Akilbekova, Dana; Bratlie, Kaitlin M.; Abraham, Thomas

The collagenous capsule formed around an implant will ultimately determine the nature of its in vivo fate. To provide a better understanding of how surface modifications can alter the collagen orientation and composition in the fibrotic capsule, we used second harmonic generation (SHG) microscopy to evaluate collagen organization and structure generated in mice subcutaneously injected with chemically functionalized polystyrene particles. SHG is sensitive to the orientation of a molecule, making it a powerful tool for measuring the alignment of collagen fibers. Additionally, SHG arises from the second order susceptibility of the interrogated molecule in response to the electric field. Variationmore » in these tensor components distinguishes different molecular sources of SHG, providing collagen type specificity. Here, we demonstrated the ability of SHG to differentiate collagen type I and type III quantitatively and used this method to examine fibrous capsules of implanted polystyrene particles. Data presented in this work shows a wide range of collagen fiber orientations and collagen compositions in response to surface functionalized polystyrene particles. Dimethylamino functionalized particles were able to form a thin collagenous matrix resembling healthy skin. These findings have the potential to improve the fundamental understanding of how material properties influence collagen organization and composition quantitatively.« less

Excitons in Cuprous Oxide: Photoionization and Other Multiphoton Processes

NASA Astrophysics Data System (ADS)

Frazer, Nicholas Laszlo

In cuprous oxide (Cu2O), momentum from the absorption of two infrared photons to make an orthoexciton is conserved and detected through the photon component of a resulting mixed exciton/photon (quadrupole exciton polariton) state. I demonstrated that this process, which actually makes the photon momentum more precisely defined, is disrupted by photoionization of excitons. Some processes are known to affect exciton propagation in both the pump and exciton stages, such as phonon emission, exciton-exciton (Auger) scattering, and third harmonic generation. These processes alone were not able to explain all observed losses of excitons or all detected scattering products, which lead me to design an optical pump-probe experiment to measure the exciton photoionization cross section, which is (3.9+/-0.2) x 10-22 m2. This dissertation describes the synthesis of cuprous oxide crystals using oxidation of copper, crystallization from melt with the optical floating zone method, and annealing. The cuprous oxide crystals were characterized using time and space resolved luminescence, leading to the discovery of new defect properties. Selection rules and overall efficiency of third harmonic generation in these crystals were characterized. Exciton photoionization was demonstrated through the depletion of polariton luminescence by an optical probe, the production of phonon linked luminescence as a scattering product, temporal delay of the probe, and time resolved luminescence. The results are integrated with the traditional dynamical model of exciton densities. An additional investigation of copper/cuprous oxide/gold photovoltaic devices is appended.

Mover Position Detection for PMTLM Based on Linear Hall Sensors through EKF Processing

PubMed Central

Yan, Leyang; Zhang, Hui; Ye, Peiqing

2017-01-01

Accurate mover position is vital for a permanent magnet tubular linear motor (PMTLM) control system. In this paper, two linear Hall sensors are utilized to detect the mover position. However, Hall sensor signals contain third-order harmonics, creating errors in mover position detection. To filter out the third-order harmonics, a signal processing method based on the extended Kalman filter (EKF) is presented. The limitation of conventional processing method is first analyzed, and then EKF is adopted to detect the mover position. In the EKF model, the amplitude of the fundamental component and the percentage of the harmonic component are taken as state variables, and they can be estimated based solely on the measured sensor signals. Then, the harmonic component can be calculated and eliminated. The proposed method has the advantages of faster convergence, better stability and higher accuracy. Finally, experimental results validate the effectiveness and superiority of the proposed method. PMID:28383505

A method for measuring the nonlinear response in dielectric spectroscopy through third harmonics detection.

PubMed

Thibierge, C; L'Hôte, D; Ladieu, F; Tourbot, R

2008-10-01

We present a high sensitivity method allowing the measurement of the nonlinear dielectric susceptibility of an insulating material at finite frequency. It has been developed for the study of dynamic heterogeneities in supercooled liquids using dielectric spectroscopy at frequencies 0.05 Hz < or = f < or = 3x10(4) Hz. It relies on the measurement of the third harmonics component of the current flowing out of a capacitor. We first show that standard laboratory electronics (amplifiers and voltage sources) nonlinearities lead to limits on the third harmonics measurements that preclude reaching the level needed by our physical goal, a ratio of the third harmonics to the fundamental signal about 10(-7). We show that reaching such a sensitivity needs a method able to get rid of the nonlinear contributions both of the measuring device (lock-in amplifier) and of the excitation voltage source. A bridge using two sources fulfills only the first of these two requirements, but allows to measure the nonlinearities of the sources. Our final method is based on a bridge with two plane capacitors characterized by different dielectric layer thicknesses. It gets rid of the source and amplifier nonlinearities because in spite of a strong frequency dependence of the capacitor impedance, it is equilibrated at any frequency. We present the first measurements of the physical nonlinear response using our method. Two extensions of the method are suggested.

3D imaging of hematoxylin and eosin stained thick tissues with a sub-femtoliter resolution by using Cr:forsterite-laser-based nonlinear microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Kao, Chien-Ting; Wei, Ming-Liang; Liao, Yi-Hua; Sun, Chi-Kuang

2017-02-01

Intraoperative assessment of excision tissues during cancer surgery is clinically important. The assessment is used to be guided by the examination for residual tumor with frozen pathology, while it is time consuming for preparation and is with low accuracy for diagnosis. Recently, reflection confocal microscopy (RCM) and nonlinear microscopy (NLM) were demonstrated to be promising methods for surgical border assessment. Intraoperative RCM imaging may enable detection of residual tumor directly on skin cancers patients during Mohs surgery. The assessment of benign and malignant breast pathologies in fresh surgical specimens was demonstrated by NLM. Without using hematoxylin and eosin (H and E) that are common dyes for histopathological diagnosis, RCM was proposed to image in vivo by using aluminum chloride for nuclear contrast on surgical wounds directly, while NLM was proposed to detect two photon fluorescence nuclear contrast from acrdine orange staining. In this paper, we propose and demonstrate 3D imaging of H and E stained thick tissues with a sub-femtoliter resolution by using Cr:forsterite-laser-based NLM. With a 1260 nm femtosecond Cr:forsterite laser as the excitation source, the hematoxylin will strongly enhance the third-harmonic generation (THG) signals, while eosin will illuminate strong fluorescence under three photon absorption. Compared with previous works, the 1260 nm excitation light provide high penetration and low photodamage to the exercised tissues so that the possibility to perform other follow-up examination will be preserved. The THG and three-photon process provides high nonlinearity so that the super resolution in 3D is now possible. The staining and the contrast of the imaging is also fully compatible with the current clinical standard on frozen pathology thus facilitate the rapid intraoperative assessment of excision tissues. This work is sponsored by National Health Research Institutes and supported by National Taiwan University Hospital.

Clinical Nonlinear Laser Imaging of Human Skin: A Review

PubMed Central

Pavone, Francesco Saverio

2014-01-01

Nonlinear optical microscopy has the potential of being used in vivo as a noninvasive imaging modality for both epidermal and dermal imaging. This paper reviews the capabilities of nonlinear microscopy as a noninvasive high-resolution tool for clinical skin inspection. In particular, we show that two-photon fluorescence microscopy can be used as a diagnostic tool for characterizing epidermal layers by means of a morphological examination. Additional functional information on the metabolic state of cells can be provided by measuring the fluorescence decay of NADH. This approach allows differentiating epidermal layers having different structural and cytological features and has the potential of diagnosing pathologies in a very early stage. Regarding therapy follow-up, we demonstrate that nonlinear microscopy could be successfully used for monitoring the effect of a treatment. In particular, combined two-photon fluorescence and second-harmonic generation microscopy were used in vivo for monitoring collagen remodeling after microablative fractional laser resurfacing and for quantitatively monitoring psoriasis on the basis of the morphology of epidermal cells and dermal papillae. We believe that the described microscopic modalities could find in the near future a stable place in a clinical dermatological setting for quantitative diagnostic purposes and as a monitoring method for various treatments. PMID:25250337

Optical tweezers and multiphoton microscopies integrated photonic tool for mechanical and biochemical cell processes studies

NASA Astrophysics Data System (ADS)

de Thomaz, A. A.; Faustino, W. M.; Fontes, A.; Fernandes, H. P.; Barjas-Castro, M. d. L.; Metze, K.; Giorgio, S.; Barbosa, L. C.; Cesar, C. L.

2007-09-01

The research in biomedical photonics is clearly evolving in the direction of the understanding of biological processes at the cell level. The spatial resolution to accomplish this task practically requires photonics tools. However, an integration of different photonic tools and a multimodal and functional approach will be necessary to access the mechanical and biochemical cell processes. This way we can observe mechanicaly triggered biochemical events or biochemicaly triggered mechanical events, or even observe simultaneously mechanical and biochemical events triggered by other means, e.g. electricaly. One great advantage of the photonic tools is its easiness for integration. Therefore, we developed such integrated tool by incorporating single and double Optical Tweezers with Confocal Single and Multiphoton Microscopies. This system can perform 2-photon excited fluorescence and Second Harmonic Generation microscopies together with optical manipulations. It also can acquire Fluorescence and SHG spectra of specific spots. Force, elasticity and viscosity measurements of stretched membranes can be followed by real time confocal microscopies. Also opticaly trapped living protozoas, such as leishmania amazonensis. Integration with CARS microscopy is under way. We will show several examples of the use of such integrated instrument and its potential to observe mechanical and biochemical processes at cell level.

Nonlinear microscopy of lipid storage and fibrosis in muscle and liver tissues of mice fed high-fat diets

NASA Astrophysics Data System (ADS)

Brackmann, Christian; Gabrielsson, Britt; Svedberg, Fredrik; Holmäng, Agneta; Sandberg, Ann-Sofie; Enejder, Annika

2010-11-01

Hallmarks of high-fat Western diet intake, such as excessive lipid accumulation in skeletal muscle and liver as well as liver fibrosis, are investigated in tissues from mice using nonlinear microscopy, second harmonic generation (SHG), and coherent anti-Stokes Raman scattering (CARS), supported by conventional analysis methods. Two aspects are presented; intake of standard chow versus Western diet, and a comparison between two high-fat Western diets of different polyunsaturated lipid content. CARS microscopy images of intramyocellular lipid droplets in muscle tissue show an increased amount for Western diet compared to standard diet samples. Even stronger diet impact is found for liver samples, where combined CARS and SHG microscopy visualize clear differences in lipid content and collagen fiber development, the latter indicating nonalcoholic fatty liver disease (NAFLD) and steatohepatitis induced at a relatively early stage for Western diet. Characteristic for NAFLD, the fibrous tissue-containing lipids accumulate in larger structures. This is also observed in CARS images of liver samples from two Western-type diets of different polyunsaturated lipid contents. In summary, nonlinear microscopy has strong potential (further promoted by technical advances toward clinical use) for detection and characterization of steatohepatitis already in its early stages.

Nonlinear microscopy of lipid storage and fibrosis in muscle and liver tissues of mice fed high-fat diets.

PubMed

Brackmann, Christian; Gabrielsson, Britt; Svedberg, Fredrik; Holmaang, Agneta; Sandberg, Ann-Sofie; Enejder, Annika

2010-01-01

Hallmarks of high-fat Western diet intake, such as excessive lipid accumulation in skeletal muscle and liver as well as liver fibrosis, are investigated in tissues from mice using nonlinear microscopy, second harmonic generation (SHG), and coherent anti-Stokes Raman scattering (CARS), supported by conventional analysis methods. Two aspects are presented; intake of standard chow versus Western diet, and a comparison between two high-fat Western diets of different polyunsaturated lipid content. CARS microscopy images of intramyocellular lipid droplets in muscle tissue show an increased amount for Western diet compared to standard diet samples. Even stronger diet impact is found for liver samples, where combined CARS and SHG microscopy visualize clear differences in lipid content and collagen fiber development, the latter indicating nonalcoholic fatty liver disease (NAFLD) and steatohepatitis induced at a relatively early stage for Western diet. Characteristic for NAFLD, the fibrous tissue-containing lipids accumulate in larger structures. This is also observed in CARS images of liver samples from two Western-type diets of different polyunsaturated lipid contents. In summary, nonlinear microscopy has strong potential (further promoted by technical advances toward clinical use) for detection and characterization of steatohepatitis already in its early stages.

Label-free imaging of brain and brain tumor specimens with combined two-photon excited fluorescence and second harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Jiang, Liwei; Wang, Xingfu; Wu, Zanyi; Du, Huiping; Wang, Shu; Li, Lianhuang; Fang, Na; Lin, Peihua; Chen, Jianxin; Kang, Dezhi; Zhuo, Shuangmu

2017-10-01

Label-free imaging techniques are gaining acceptance within the medical imaging field, including brain imaging, because they have the potential to be applied to intraoperative in situ identifications of pathological conditions. In this paper, we describe the use of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) microscopy in combination for the label-free detection of brain and brain tumor specimens; gliomas. Two independently detecting channels were chosen to subsequently collect TPEF/SHG signals from the specimen to increase TPEF/SHG image contrasts. Our results indicate that the combined TPEF/SHG microscopic techniques can provide similar rat brain structural information and produce a similar resolution like conventional H&E staining in neuropathology; including meninges, cerebral cortex, white-matter structure corpus callosum, choroid plexus, hippocampus, striatum, and cerebellar cortex. It can simultaneously detect infiltrating human brain tumor cells, the extracellular matrix collagen fiber of connective stroma within brain vessels and collagen depostion in tumor microenvironments. The nuclear-to-cytoplasmic ratio and collagen content can be extracted as quantitative indicators for differentiating brain gliomas from healthy brain tissues. With the development of two-photon fiberscopes and microendoscope probes and their clinical applications, the combined TPEF and SHG microcopy may become an important multimodal, nonlinear optical imaging approach for real-time intraoperative histological diagnostics of residual brain tumors. These occur in various brain regions during ongoing surgeries through the method of simultaneously identifying tumor cells, and the change of tumor microenvironments, without the need for the removal biopsies and without the need for tissue labelling or fluorescent markers.

Second-harmonic generation microscopy used to evaluate the effect of the dimethyl sulfoxide in the cryopreservation process in collagen fibers of differentiated chondrocytes

NASA Astrophysics Data System (ADS)

Andreoli-Risso, M. F.; Duarte, A. S. S.; Ribeiro, T. B.; Bordeaux-Rego, P.; Luzo, A.; Baratti, M. O.; Adur, J.; de Thomaz, A. A.; Pelegati, V. B.; Carvalho, H. F.; Cesar, C. L.; Kharmadayan, P.; Costa, F. F.; Olalla-Saad, S. T.

2012-03-01

Cartilaginous lesions are a significant public health problem and the use of adult stem cells represents a promising therapy for this condition. Cryopreservation confers many advantages for practitioners engaged in cell-based therapies. However, conventional slow freezing has always been associated with damage and mortality due to intracellular ice formation, cryoprotectant toxicity, and dehydration. The aim of this work is to observe the effect of the usual Dimethyl Sulfoxide (DMSO) cryopreservation process on the architecture of the collagen fiber network of chondrogenic cells from mesenchymal stem cells by Second Harmonic Generation (SHG) microscopy. To perform this study we used Mesenchymal Stem Cells (MSC) derived from adipose tissue which presents the capacity to differentiate into other lineages such as osteogenic, adipogenic and chondrogenic lineages. Mesenchymal stem cells obtained after liposuction were isolated digested by collagenase type I and characterization was carried out by differentiation of mesodermic lineages, and flow cytometry using specific markers. The isolated MSCs were cryopreserved by the DMSO technique and the chondrogenic differentiation was carried out using the micromass technique. We then compared the cryopreserved vs non-cryopreserved collagen fibers which are naturally formed during the differentiation process. We observed that noncryopreserved MSCs presented a directional trend in the collagen fibers formed which was absent in the cryopreserved MSCs. We confirmed this trend quantitatively by the aspect ratio obtained by Fast Fourier Transform which was 0.76 for cryopreserved and 0.52 for non-cryopreserved MSCs, a statistical significant difference.

Second-harmonic generation and fluorescence lifetime imaging microscopy through a rodent mammary imaging window

NASA Astrophysics Data System (ADS)

Young, Pamela A.; Nazir, Muhammad; Szulczewski, Michael J.; Keely, Patricia J.; Eliceiri, Kevin W.

2012-03-01

Tumor-Associated Collagen Signatures (TACS) have been identified that manifest in specific ways during breast tumor progression and that correspond to patient outcome. There are also compelling metabolic changes associated with carcinoma invasion and progression. We have characterized the difference in the autofluorescent properties of metabolic co-factors, NADH and FAD, between normal and carcinoma breast cell lines. Also, we have shown in vitro that increased collagen density alters metabolic genes which are associated with glycolysis and leads to a more invasive phenotype. Establishing the relationship between collagen density, cellular metabolism, and metastasis in physiologically relevant cancer models is crucial for developing cancer therapies. To study cellular metabolism with respect to collagen density in vivo, we use multiphoton fluorescence excitation microscopy (MPM) in conjunction with a rodent mammary imaging window implanted in defined mouse cancer models. These models are ideal for the study of collagen changes in vivo, allowing determination of corresponding metabolic changes in breast cancer invasion and progression. To measure cellular metabolism, we collect fluorescence lifetime (FLIM) signatures of NADH and FAD, which are known to change based on the microenvironment of the cells. Additionally, MPM systems are capable of collecting second harmonic generation (SHG) signals which are a nonlinear optical property of collagen. Therefore, MPM, SHG, and FLIM are powerful tools with great potential for characterizing key features of breast carcinoma in vivo. Below we present the current efforts of our collaborative group to develop intravital approaches based on these imaging techniques to look at defined mouse mammary models.

Tailored semiconductors for high-harmonic optoelectronics.

PubMed

Sivis, Murat; Taucer, Marco; Vampa, Giulio; Johnston, Kyle; Staudte, André; Naumov, Andrei Yu; Villeneuve, D M; Ropers, Claus; Corkum, P B

2017-07-21

The advent of high-harmonic generation in gases 30 years ago set the foundation for attosecond science and facilitated ultrafast spectroscopy in atoms, molecules, and solids. We explore high-harmonic generation in the solid state by means of nanostructured and ion-implanted semiconductors. We use wavelength-selective microscopic imaging to map enhanced harmonic emission and show that the generation medium and the driving field can be locally tailored in solids by modifying the chemical composition and morphology. This enables the control of high-harmonic technology within precisely engineered solid targets. We demonstrate customized high-harmonic wave fields with wavelengths down to 225 nanometers (ninth-harmonic order of 2-micrometer laser pulses) and present an integrated Fresnel zone plate target in silicon, which leads to diffraction-limited self-focusing of the generated harmonics down to 1-micrometer spot sizes. Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

Direct laser interference patterning of ophthalmic polydimethylsiloxane (PDMS) polymers

NASA Astrophysics Data System (ADS)

Sola, D.; Lavieja, C.; Orera, A.; Clemente, M. J.

2018-07-01

The inscription of diffractive elements in ophthalmic polymers and ocular tissues to induce refractive index changes is of great interest in the fields of Optics and Ophthalmology. In this work fabrication of linear periodic patterns in polydimethylsiloxane (PDMS) intraocular lenses by means of the direct laser interference patterning (DLIP) technique was studied. A Q-Switch Nd:YAG laser coupled to second and third harmonic modules emitting linearly polarized 4 ns pulses at 355 nm with 20 Hz repetition rate was used as the laser source. Laser processing parameters were modified to produce the linear patterns. Processed samples were characterized by means of optical confocal microscopy, Scanning Electron Microscopy SEM, Energy Dispersive X-ray Spectroscopy EDX, Attenuated Total Reflectance-Infrared Spectroscopy ATR-FTIR, and Raman Spectroscopy. Depending on the laser parameters both photo-thermal and photo-chemical damage were observed in the DLIP irradiated areas. Finally, diffractive techniques were used to characterize the diffraction gratings inscribed in the samples resulting in a refractive index change of 1.9 × 10-2 under illumination of a 632.8 nm He-Ne laser.

Identification of normal and cancerous human colorectal muscularis propria by multiphoton microscopy in different sections

NASA Astrophysics Data System (ADS)

Zhou, Yi; Chen, Zhifen; Kang, Deyong; li, Lianhuang; Zhuo, Shuangmu; Zhu, Xiaoqin; Guan, Guoxian; Chen, Jianxin

2016-01-01

Multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) as a potential diagnostic tool is attractive. MPM can effectively provide information about morphological and biochemical changes in biological tissues at the molecular level. In this paper, we attempt to identify normal and cancerous human colorectal muscularis propria by multiphoton microscopy in different sections (both in transverse and longitudinal sections). The results show that MPM can display different microstructure changes in the transverse and longitudinal sections of colorectal muscularis propria. MPM also can quantitatively describe the alteration of collagen content between normal and cancerous muscle layers. These are important pathological findings that MPM images can bring more detailed complementary information about tissue architecture and cell morphology through observing the transverse and longitudinal sections of colorectal muscularis propria. This work demonstrates that MPM can be better for identifying the microstructural characteristics of normal and cancerous human colorectal muscularis propria in different sections.

High-resolution measurements of the multilayer ultra-structure of articular cartilage and their translational potential

PubMed Central

2014-01-01

Current musculoskeletal imaging techniques usually target the macro-morphology of articular cartilage or use histological analysis. These techniques are able to reveal advanced osteoarthritic changes in articular cartilage but fail to give detailed information to distinguish early osteoarthritis from healthy cartilage, and this necessitates high-resolution imaging techniques measuring cells and the extracellular matrix within the multilayer structure of articular cartilage. This review provides a comprehensive exploration of the cellular components and extracellular matrix of articular cartilage as well as high-resolution imaging techniques, including magnetic resonance image, electron microscopy, confocal laser scanning microscopy, second harmonic generation microscopy, and laser scanning confocal arthroscopy, in the measurement of multilayer ultra-structures of articular cartilage. This review also provides an overview for micro-structural analysis of the main components of normal or osteoarthritic cartilage and discusses the potential and challenges associated with developing non-invasive high-resolution imaging techniques for both research and clinical diagnosis of early to late osteoarthritis. PMID:24946278

Alignment-free, all-spliced fiber laser source for CARS microscopy based on four-wave-mixing.

PubMed

Baumgartl, Martin; Gottschall, Thomas; Abreu-Afonso, Javier; Díez, Antonio; Meyer, Tobias; Dietzek, Benjamin; Rothhardt, Manfred; Popp, Jürgen; Limpert, Jens; Tünnermann, Andreas

2012-09-10

An environmentally-stable low-repetition rate fiber oscillator is developed to produce narrow-bandwidth pulses with several tens of picoseconds duration. Based on this oscillator an alignment-free all-fiber laser for multi-photon microscopy is realized using in-fiber frequency conversion based on four-wave-mixing. Both pump and Stokes pulses for coherent anti-Stokes Raman scattering (CARS) microscopy are readily available from one fiber end, intrinsically overlapped in space and time, which drastically simplifies the experimental handling for the user. The complete laser setup is mounted on a home-built laser scanning microscope with small footprint. High-quality multimodal microscope images of biological tissue are presented probing the CH-stretching resonance of lipids at an anti-Stokes Raman-shift of 2845 cm(-1) and second-harmonic generation of collagen. Due to its simplicity, compactness, maintenance-free operation, and ease-of-use the presented low-cost laser is an ideal source for bio-medical applications outside laser laboratories and in particular inside clinics.

Experimental demonstration of efficient and robust second harmonic generation using the adiabatic temperature gradient method

NASA Astrophysics Data System (ADS)

Dimova, E.; Steflekova, V.; Karatodorov, S.; Kyoseva, E.

2018-03-01

We propose a way of achieving efficient and robust second-harmonic generation. The technique proposed is similar to the adiabatic population transfer in a two-state quantum system with crossing energies. If the phase mismatching changes slowly, e.g., due to a temperature gradient along the crystal, and makes the phase match for second-harmonic generation to occur, then the energy would be converted adiabatically to the second harmonic. As an adiabatic technique, the second-harmonic generation scheme presented is stable to variations in the crystal parameters, as well as in the input light, crystal length, input intensity, wavelength and angle of incidence.

Nonlinear dynamics of nanoscale systems

NASA Astrophysics Data System (ADS)

Hodas, Nathan Oken

This work builds theoretical tools to better understand nanoscale systems, and it ex- plores experimental techniques to probe nanoscale dynamics using nonlinear optical microscopy. In both the theory and experiment, this work harnesses nonlinearity to explore new boundaries in the ongoing attempts to understand the amazing world that is much smaller than we can see. In particular, the first part of this work proves the upper-bounds on the number and quality of oscillations when the sys- tem in question is homogeneously driven and has discrete states, a common way of describing nanoscale motors and chemical systems, although it has application to networked systems in general. The consequences of this limit are explored in the context of chemical clocks and limit cycles. This leads to the analysis of sponta- neous oscillations in GFPmut2, where we postulate that the oscillations must be due to coordinated rearrangement of the beta-barrel. Next, we utilize nonlinear optics to probe the constituent structures of zebrafish muscle. By comparing experimental observations with computational models, we show how second harmonic generation differs from fluorescence for confocal imaging. We use the wavelength dependence of the second harmonic generation conversion efficiency to extract information about the microscopic organization of muscle fibers, using the coherent nature of second ix harmonic generation as an analytical probe. Finally, existing experiments have used a related technique, sum-frequency generation, to directly probe the dynamics of free OH bonds at the water-vapor boundary. Using molecular dynamic simulations of the water surface and by designating surface-sensitive free OH bonds on the water surface, many aspects of the sum-frequency generation measurements were calcu- lated and compared with those inferred from experiment. The method utilizes results available from independent IR and Raman experiments to obtain some of the needed quantities, rather than calculating them ab initio. The results provide insight into the microscopic dynamics at the air-water interface and have useful application in the field of on-water catalysis.

Three-Dimensional Geometry of Collagenous Tissues by Second Harmonic Polarimetry.

PubMed

Reiser, Karen; Stoller, Patrick; Knoesen, André

2017-06-01

Collagen is a biological macromolecule capable of second harmonic generation, allowing label-free detection in tissues; in addition, molecular orientation can be determined from the polarization dependence of the second harmonic signal. Previously we reported that in-plane orientation of collagen fibrils could be determined by modulating the polarization angle of the laser during scanning. We have now extended this method so that out-of-plane orientation angles can be determined at the same time, allowing visualization of the 3-dimensional structure of collagenous tissues. This approach offers advantages compared with other methods for determining out-of-plane orientation. First, the orientation angles are directly calculated from the polarimetry data obtained in a single scan, while other reported methods require data from multiple scans, use of iterative optimization methods, application of fitting algorithms, or extensive post-optical processing. Second, our method does not require highly specialized instrumentation, and thus can be adapted for use in almost any nonlinear optical microscopy setup. It is suitable for both basic and clinical applications. We present three-dimensional images of structurally complex collagenous tissues that illustrate the power of such 3-dimensional analyses to reveal the architecture of biological structures.

Three-Dimensional Geometry of Collagenous Tissues by Second Harmonic Polarimetry

DOE Office of Scientific and Technical Information (OSTI.GOV)

Reiser, Karen; Stoller, Patrick; Knoesen, André

Collagen is a biological macromolecule capable of second harmonic generation, allowing label-free detection in tissues; in addition, molecular orientation can be determined from the polarization dependence of the second harmonic signal. Previously we reported that in-plane orientation of collagen fibrils could be determined by modulating the polarization angle of the laser during scanning. We have now extended this method so that out-of-plane orientation angles can be determined at the same time, allowing visualization of the 3-dimensional structure of collagenous tissues. This approach offers advantages compared with other methods for determining out-of-plane orientation. First, the orientation angles are directly calculated frommore » the polarimetry data obtained in a single scan, while other reported methods require data from multiple scans, use of iterative optimization methods, application of fitting algorithms, or extensive post-optical processing. Second, our method does not require highly specialized instrumentation, and thus can be adapted for use in almost any nonlinear optical microscopy setup. It is suitable for both basic and clinical applications. We present three-dimensional images of structurally complex collagenous tissues that illustrate the power of such 3-dimensional analyses to reveal the architecture of biological structures.« less

Efficient second-harmonic imaging of collagen in histological slides using Bessel beam excitation

NASA Astrophysics Data System (ADS)

Vuillemin, Nelly; Mahou, Pierre; Débarre, Delphine; Gacoin, Thierry; Tharaux, Pierre-Louis; Schanne-Klein, Marie-Claire; Supatto, Willy; Beaurepaire, Emmanuel

2016-07-01

Second-harmonic generation (SHG) is the most specific label-free indicator of collagen accumulation in widespread pathologies such as fibrosis, and SHG-based measurements hold important potential for biomedical analyses. However, efficient collagen SHG scoring in histological slides is hampered by the limited depth-of-field of usual nonlinear microscopes relying on focused Gaussian beam excitation. In this work we analyze theoretically and experimentally the use of Bessel beam excitation to address this issue. Focused Bessel beams can provide an axially extended excitation volume for nonlinear microscopy while preserving lateral resolution. We show that shaping the focal volume has consequences on signal level and scattering directionality in the case of coherent signals (such as SHG) which significantly differ from the case of incoherent signals (two-photon excited fluorescence, 2PEF). We demonstrate extended-depth SHG-2PEF imaging of fibrotic mouse kidney histological slides. Finally, we show that Bessel beam excitation combined with spatial filtering of the harmonic light in wave vector space can be used to probe collagen accumulation more efficiently than the usual Gaussian excitation scheme. These results open the way to SHG-based histological diagnoses.

Three-Dimensional Geometry of Collagenous Tissues by Second Harmonic Polarimetry

DOE PAGES

Reiser, Karen; Stoller, Patrick; Knoesen, André

2017-06-01

Collagen is a biological macromolecule capable of second harmonic generation, allowing label-free detection in tissues; in addition, molecular orientation can be determined from the polarization dependence of the second harmonic signal. Previously we reported that in-plane orientation of collagen fibrils could be determined by modulating the polarization angle of the laser during scanning. We have now extended this method so that out-of-plane orientation angles can be determined at the same time, allowing visualization of the 3-dimensional structure of collagenous tissues. This approach offers advantages compared with other methods for determining out-of-plane orientation. First, the orientation angles are directly calculated frommore » the polarimetry data obtained in a single scan, while other reported methods require data from multiple scans, use of iterative optimization methods, application of fitting algorithms, or extensive post-optical processing. Second, our method does not require highly specialized instrumentation, and thus can be adapted for use in almost any nonlinear optical microscopy setup. It is suitable for both basic and clinical applications. We present three-dimensional images of structurally complex collagenous tissues that illustrate the power of such 3-dimensional analyses to reveal the architecture of biological structures.« less

Efficient second-harmonic imaging of collagen in histological slides using Bessel beam excitation

PubMed Central

Vuillemin, Nelly; Mahou, Pierre; Débarre, Delphine; Gacoin, Thierry; Tharaux, Pierre-Louis; Schanne-Klein, Marie-Claire; Supatto, Willy; Beaurepaire, Emmanuel

2016-01-01

Second-harmonic generation (SHG) is the most specific label-free indicator of collagen accumulation in widespread pathologies such as fibrosis, and SHG-based measurements hold important potential for biomedical analyses. However, efficient collagen SHG scoring in histological slides is hampered by the limited depth-of-field of usual nonlinear microscopes relying on focused Gaussian beam excitation. In this work we analyze theoretically and experimentally the use of Bessel beam excitation to address this issue. Focused Bessel beams can provide an axially extended excitation volume for nonlinear microscopy while preserving lateral resolution. We show that shaping the focal volume has consequences on signal level and scattering directionality in the case of coherent signals (such as SHG) which significantly differ from the case of incoherent signals (two-photon excited fluorescence, 2PEF). We demonstrate extended-depth SHG-2PEF imaging of fibrotic mouse kidney histological slides. Finally, we show that Bessel beam excitation combined with spatial filtering of the harmonic light in wave vector space can be used to probe collagen accumulation more efficiently than the usual Gaussian excitation scheme. These results open the way to SHG-based histological diagnoses. PMID:27435390

Intraoperative assisting diagnosis of esophageal submucosal cancer using multiphoton microscopy

NASA Astrophysics Data System (ADS)

Zeng, Yaping; Xu, Jian; Zhou, Qun; Kang, Deyong; Zhuo, Shuangmu; Zhu, Xiaoqin; Lin, Jiangbo; Chen, Jianxin

2018-07-01

Multiphoton microscopy (MPM) based on two-photon excited fluorescence and second harmonic generation can achieve high-resolution images of biological tissues at the cellular and subcellular levels. In this work, we used MPM imaging of intraoperative hematoxylin and eosin (H&E)-stained frozen sections (FSs) of esophagus to explore whether MPM can provide complementary information to the auxiliary diagnosis of esophageal submucosal cancer during the intraoperative period. It was found that MPM has the ability not only to clearly reveal biological tissue microstructure and its morphological changes, but can reveal information not distinguishable in H&E-stained images. The complementary information of nonlinear spectral analysis, orientation and morphology changes in the collagen showed that MPM has important accessory diagnostic value for the differential diagnosis of submucosal carcinoma of the esophagus during the intraoperative period.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Newman, Justin A.; Schmitt, Paul D.; Toth, Scott J.

Here in this paper we demonstrate the use of second harmonic generation (SHG) microscopy-guided synchrotron powder X-ray diffraction (PXRD) for the detection of trace crystalline active pharmaceutical ingredients in a common polymer blend. The combined instrument is capable of detecting 100 ppm crystalline ritonavir in an amorphous hydroxypropyl methylcellulose matrix with a high signal-to-noise ratio (>5000). The high spatial resolution afforded by SHG microscopy allows for the use of a minibeam collimator to reduce the total volume of material probed by synchrotron PXRD. The reduction in probed volume results in reduced background from amorphous material. The ability to detect lowmore » crystalline loading has the potential to improve measurements in the formulation pipeline for pharmaceutical solid dispersions, for which even trace quantities of crystalline active ingredients can negatively impact the stability and bioavailability of the final drug product.« less

Exploring correlations in the CGC wave function: Odd azimuthal anisotropy

DOE PAGES

Kovner, Alex; Lublinsky, Michael; Skokov, Vladimir

2017-07-17

In this paper, we extend the color glass condensate (CGC) approach to a calculation of the double inclusive gluon production by including the high density effect in the CGC wave function of the projectile (proton). Our main result is that these effects lead to the appearance of odd harmonics in the two particle correlation C(k,p). We find that in the high momentum limit, |k|, |p| >> Q s, this results in a positive c 1{2}. Additionally when the magnitudes of the two momenta are approximately equal, |k|/|p| ≈ 1, the density effects also generate a positive third harmonic c 3{2},more » which translates into a nonvanishing v 3 when the momenta of the trigger and an associated particle are in the same momentum bin. Finally, the sign of c 3{2} becomes negative when |k|/|p| > 1.1 suggesting an interesting experimental signature.« less

Large optical second-order nonlinearity of poled WO3-TeO2 glass.

PubMed

Tanaka, K; Narazaki, A; Hirao, K

2000-02-15

Second-harmonic generation, one of the second-order nonlinear optical properties of thermally and electrically poled WO>(3)-TeO>(2) glasses, has been examined. We poled glass samples with two thicknesses (0.60 and 0.86 mm) at various temperatures to explore the effects of external electric field strength and poling temperature on second-order nonlinearity. The dependence of second-harmonic intensity on the poling temperature is maximum at a specific poling temperature. A second-order nonlinear susceptibility of 2.1 pm/V was attained for the 0.60-mm-thick glass poled at 250 degrees C. This value is fairly large compared with those for poled silica and tellurite glasses reported thus far. We speculate that the large third-order nonlinear susceptibility of WO>(3)- TeO>(2) glasses gives rise to the large second-order nonlinearity by means of a X((2)) = 3X((3)) E(dc) process.

Mode-locked Yb:YAG thin-disk oscillator with 41 µJ pulse energy at 145 W average infrared power and high power frequency conversion.

PubMed

Bauer, Dominik; Zawischa, Ivo; Sutter, Dirk H; Killi, Alexander; Dekorsy, Thomas

2012-04-23

We demonstrate the generation of 1.1 ps pulses containing more than 41 µJ of energy directly out of an Yb:YAG thin-disk without any additional amplification stages. The laser oscillator operates in ambient atmosphere with a 3.5 MHz repetition rate and 145 W of average output power at a fundamental wavelength of 1030 nm. An average output power of 91.5 W at 515 nm was obtained by frequency doubling with a conversion efficiency exceeding 65%. Third harmonic generation resulted in 34 W at 343 nm at 34% efficiency. © 2012 Optical Society of America

Zonal harmonic model of Saturn's magnetic field from Voyager 1 and 2 observations

NASA Technical Reports Server (NTRS)

Connerney, J. E. P.; Ness, N. F.; Acuna, M. H.

1982-01-01

An analysis of the magnetic field of Saturn is presented which takes into account both the Voyager 1 and 2 vector magnetic field observations. The analysis is based on the traditional spherical harmonic expansion of a scale potential to derive the magnetic field within 8 Saturn radii. A third-order zonal harmonic model fitted to Voyager 1 and 2 observations is found to be capable of predicting the magnetic field characteristics at one encounter based on those observed at another, unlike models including dipole and quadrupole terms only. The third-order model is noted to lead to significantly enhanced polar surface field intensities with respect to dipole models, and probably represents the axisymmetric part of a complex dynamo field.

High-flux soft x-ray harmonic generation from ionization-shaped few-cycle laser pulses

PubMed Central

Brahms, Christian; Gregory, Andrew; Tisch, John W. G.; Marangos, Jon P.

2018-01-01

Laser-driven high-harmonic generation provides the only demonstrated route to generating stable, tabletop attosecond x-ray pulses but has low flux compared to other x-ray technologies. We show that high-harmonic generation can produce higher photon energies and flux by using higher laser intensities than are typical, strongly ionizing the medium and creating plasma that reshapes the driving laser field. We obtain high harmonics capable of supporting attosecond pulses up to photon energies of 600 eV and a photon flux inside the water window (284 to 540 eV) 10 times higher than previous attosecond sources. We demonstrate that operating in this regime is key for attosecond pulse generation in the x-ray range and will become increasingly important as harmonic generation moves to fields that drive even longer wavelengths. PMID:29756033

Two color laser driven THz generation in clustered plasma

NASA Astrophysics Data System (ADS)

Malik, Rakhee; Uma, R.; Kumar, Pawan

2017-07-01

A scheme of terahertz (THz) generation, using nonlinear mixing of two color laser (fundamental ω1 and slightly frequency shifted second harmonic ω2 ) in clustered plasma, is investigated. The lasers exert ponderomotive force on cluster electrons and drive density perturbations at 2 ω1 and ω2-ω1 . The density perturbations beat with the oscillatory velocities to produce nonlinear current at ω2-2 ω1 , generating THz radiation. The radiation is enhanced due to cluster plasmon resonance and by phase matching introduced through a density ripple. The generation involves third order nonlinearity and does not require a magnetic field or inhomogeneity to sustain it. We report THz power conversion efficiency ˜ 10-4 at 1 μm and 0.5 μm wavelengths with intensity ˜ 3 ×1014W/cm 2 .

Quantification of collagen fiber organization in biological tissues at cellular and molecular scales using second-harmonic generation imaging

NASA Astrophysics Data System (ADS)

Ambekar Ramachandra Rao, Raghu

Collagen is the most abundant structural protein found in the human body, and is responsible for providing structure and function to tissues. Collagen molecules organize naturally into structures called fibers on the scale of the wavelength of light and lack inversion symmetry, thus allowing for the process of second harmonic generation (SHG) when exposed to intense incident light. We have developed two quantitative techniques: Fourier transform-second-harmonic generation (FT-SHG) imaging and generalized chi2 second-harmonic generation (chi2-SHG) imaging. In order to show that FT-SHG imaging can be used as a valuable diagnostic tool for real-world biological problems, we first investigate collagenase-induced injury in horse tendons. Clear differences in collagen fiber organization between normal and injured tendon are quantified. In particular, we observe that the regularly oriented organization of collagen fibers in normal tendons is disrupted in injured tendons leading to a more random organization. We also observe that FT-SHG microscopy is more sensitive in assessing tendon injury compared to the conventional polarized light microscopy. The second study includes quantifying collagen fibers in cortical bone using FT-SHG imaging and comparing it with scanning electron microscopy (SEM). Further, as an example study, we show how FT-SHG imaging could be used to quantify changes in bone structure as a function of age. Some initial work and future directions for extending FT-SHG to 3D are also discussed. The second technique, chi2-SHG imaging, takes advantage of the coherent nature of SHG and utilizes polarization to extract the second-order susceptibility (d elements) which provides information on molecular organization, i.e., it provides access to sub-diffractional changes "optically". We use chi2-SHG in combination with FT-SHG imaging to investigate a couple of biological problems. First, we quantify differences in collagen fiber organization between cornea and sclera of the eye in order to investigate their properties of transparency and opacity, respectively. We find from chi2-SHG imaging that there is no statistical difference in the values of d elements between cornea and sclera, indicating that the underlying collagen structure generating SHG from the two is similar at the level of detection of SHG microscopy. However, the difference lies in the spatial organization of these collagen fibers as observed from FT-SHG imaging. We find that cornea contains lamellae with patches of ordered and uniform diameter collagen fibers with axial order, which could be the reason for its transparent behavior. Conversely, there are no lamellae in sclera (i.e., no axial order), and fibers are thicker, denser, have inconsistent diameters, and possess relatively inhomogeneous orientations, leading to its opaque nature. We also utilized the two techniques to assess differences in stromal collagen fibers for several human breast tissue conditions: normal, hyperplasia, dysplasia, and malignant. Using FT-SHG imaging, we note differences between malignant and other pathological conditions through the metric A.I. ratio. Using generalized chi2-SHG imaging, we observe structural changes in collagen at the molecular scale, and a particular d element showed a more sensitive differentiation between breast tissue conditions, except between hyperplasia and normal/dysplasia. We also find that the trigonal symmetry (3m) is a more appropriate model to describe collagen fibers in malignant tissues as opposed to the conventionally used hexagonal symmetry (C6). Furthermore, the percentage of abnormal collagen fibers could potentially be used as a metric for differentiating breast tissue conditions. We also introduce a technique for extending chi2-SHG to fibers with curvature which is useful for generating chi2-image maps (in terms of d elements) instead of the conventional SHG intensity images. The spatial variations in d elements will provide additional information. For example, in breast cancer tissues, it may help in observing how fibers change from normal to malignant spatially, especially around region of cancerous cells. Finally, we discuss some of the interesting immediate and later future work of quantitative SHG imaging we aim to carry out in our lab. (Abstract shortened by UMI.)

Evaluation of the histological and mechanical features of tendon healing in a rabbit model with the use of second-harmonic-generation imaging and tensile testing.

PubMed

Hase, E; Sato, K; Yonekura, D; Minamikawa, T; Takahashi, M; Yasui, T

2016-11-01

This study aimed to evaluate the histological and mechanical features of tendon healing in a rabbit model with second-harmonic-generation (SHG) imaging and tensile testing. A total of eight male Japanese white rabbits were used for this study. The flexor digitorum tendons in their right leg were sharply transected, and then were repaired by intratendinous stitching. At four weeks post-operatively, the rabbits were killed and the flexor digitorum tendons in both right and left legs were excised and used as specimens for tendon healing (n = 8) and control (n = 8), respectively. Each specimen was examined by SHG imaging, followed by tensile testing, and the results of the two testing modalities were assessed for correlation. While the SHG light intensity of the healing tendon samples was significantly lower than that of the uninjured tendon samples, 2D Fourier transform SHG images showed a clear difference in collagen fibre structure between the uninjured and the healing samples, and among the healing samples. The mean intensity of the SHG image showed a moderate correlation (R 2 = 0.37) with Young's modulus obtained from the tensile testing. Our results indicate that SHG microscopy may be a potential indicator of tendon healing.Cite this article: E. Hase, K. Sato, D. Yonekura, T. Minamikawa, M. Takahashi, T. Yasui. Evaluation of the histological and mechanical features of tendon healing in a rabbit model with the use of second-harmonic-generation imaging and tensile testing. Bone Joint Res 2016;5:577-585. DOI: 10.1302/2046-3758.511.BJR-2016-0162.R1. © 2016 Yasui et al.

3D texture analysis for classification of second harmonic generation images of human ovarian cancer

NASA Astrophysics Data System (ADS)

Wen, Bruce; Campbell, Kirby R.; Tilbury, Karissa; Nadiarnykh, Oleg; Brewer, Molly A.; Patankar, Manish; Singh, Vikas; Eliceiri, Kevin. W.; Campagnola, Paul J.

2016-10-01

Remodeling of the collagen architecture in the extracellular matrix (ECM) has been implicated in ovarian cancer. To quantify these alterations we implemented a form of 3D texture analysis to delineate the fibrillar morphology observed in 3D Second Harmonic Generation (SHG) microscopy image data of normal (1) and high risk (2) ovarian stroma, benign ovarian tumors (3), low grade (4) and high grade (5) serous tumors, and endometrioid tumors (6). We developed a tailored set of 3D filters which extract textural features in the 3D image sets to build (or learn) statistical models of each tissue class. By applying k-nearest neighbor classification using these learned models, we achieved 83-91% accuracies for the six classes. The 3D method outperformed the analogous 2D classification on the same tissues, where we suggest this is due the increased information content. This classification based on ECM structural changes will complement conventional classification based on genetic profiles and can serve as an additional biomarker. Moreover, the texture analysis algorithm is quite general, as it does not rely on single morphological metrics such as fiber alignment, length, and width but their combined convolution with a customizable basis set.

Quantification of focal adhesion dynamics of cell movement based on cell-induced collagen matrix deformation using second-harmonic generation microscopy.

PubMed

Kang, Yong Guk; Jang, Hwanseok; Yang, Taeseok Daniel; Notbohm, Jacob; Choi, Youngwoon; Park, Yongdoo; Kim, Beop-Min

2018-06-01

Mechanical interactions of living cells with the surrounding environment via focal adhesion (FA) in three dimensions (3-D) play a key role in dynamic biological events, such as tissue regeneration, wound healing, and cancer invasion. Recently, several methods for observing 3-D cell-extracellular matrix (ECM) interactions have been reported, lacking solid and quantitative analysis on the dynamics of the physical interaction between the cell and the ECM. We measured the submicron displacements of ECM deformation in 3-D due to protrusion-retraction dynamics during cell migration, using second-harmonic generation without labeling the matrix structures. We then quantitatively analyzed the mechanical deformation between the ECM and the cells based on spatiotemporal volumetric correlations. The greatest deformations within the collagen matrix were found to occur at sites of colocalization of the FA site-related proteins vinculin and actin, which confirms that FA sites play a critical role in living cells within the ECM as a point for adhesion, traction, and migration. We believe that this modality can be used in studies of cell-ECM interaction during angiogenesis, wound healing, and metastasis. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Second harmonic generation imaging as a potential tool for staging pregnancy and predicting preterm birth

NASA Astrophysics Data System (ADS)

Akins, Meredith L.; Luby-Phelps, Katherine; Mahendroo, Mala

2010-03-01

We use second harmonic generation (SHG) microscopy to assess changes in collagen structure of murine cervix during cervical remodeling of normal pregnancy and in a preterm birth model. Visual inspection of SHG images revealed substantial changes in collagen morphology throughout normal gestation. SHG images collected in both the forward and backward directions were analyzed quantitatively for changes in overall mean intensity, forward to backward intensity ratio, collagen fiber size, and porosity. Changes in mean SHG intensity and intensity ratio take place in early pregnancy, suggesting that submicroscopic changes in collagen fibril size and arrangement occur before macroscopic changes become evident. Fiber size progressively increased from early to late pregnancy, while pores between collagen fibers became larger and farther apart. Analysis of collagen features in premature cervical remodeling show that changes in collagen structure are dissimilar from normal remodeling. The ability to quantify multiple morphological features of collagen that characterize normal cervical remodeling and distinguish abnormal remodeling in preterm birth models supports future studies aimed at development of SHG endoscopic devices for clinical assessment of collagen changes during pregnancy in women and for predicting risk of preterm labor which occurs in 12.5% of all pregnancies.

Harmonic growth of spherical Rayleigh-Taylor instability in weakly nonlinear regime

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Wanhai; LHD, Institute of Mechanics, Chinese Academy of Sciences, Beijing 100190; Chen, Yulian

Harmonic growth in classical Rayleigh-Taylor instability (RTI) on a spherical interface is analytically investigated using the method of the parameter expansion up to the third order. Our results show that the amplitudes of the first four harmonics will recover those in planar RTI as the interface radius tends to infinity compared against the initial perturbation wavelength. The initial radius dramatically influences the harmonic development. The appearance of the second-order feedback to the initial unperturbed interface (i.e., the zeroth harmonic) makes the interface move towards the spherical center. For these four harmonics, the smaller the initial radius is, the faster theymore » grow.« less

Application of organic compounds for high-order harmonic generation of ultrashort pulses

NASA Astrophysics Data System (ADS)

Ganeev, R. A.

2016-02-01

The studies of the high-order nonlinear optical properties of a few organic compounds (polyvinyl alcohol, polyethylene, sugar, coffee, and leaf) are reported. Harmonic generation in the laser-produced plasmas containing the molecules and large particles of above materials is demonstrated. These studies showed that the harmonic distributions and harmonic cutoffs from organic compound plasmas were similar to those from the graphite ablation. The characteristic feature of observed harmonic spectra was the presence of bluesided lobes near the lower-order harmonics.

Self-Action of Second Harmonic Generation and Longitudinal Temperature Gradient in Nonlinear-Optical Crystals

NASA Astrophysics Data System (ADS)

Baranov, A. I.; Konyashkin, A. V.; Ryabushkin, O. A.

2015-09-01

Model of second harmonic generation with thermal self-action was developed. Second harmonic generation temperature phase matching curves were measured and calculated for periodically polled lithium niobate crystal. Both experimental and calculated data show asymmetrical shift of temperature tuning curves with pump power.

Reflection second harmonic generation on a z -cut congruent lithium niobate crystal

NASA Astrophysics Data System (ADS)

Sono, T. J.; Scott, J. G.; Sones, C. L.; Valdivia, C. E.; Mailis, S.; Eason, R. W.; Frey, J. G.; Danos, L.

2006-11-01

Reflection second harmonic generation experiments were performed on z -cut congruent lithium niobate crystals (LiNbO3) to reveal the interfacial layer symmetry as the crystal is rotated around the z axis. To suppress the bulk contribution, the fundamental wavelength was selected to be 532nm , resulting in second harmonic generation at a wavelength within the absorption region of the crystal. The polarity of the direction of the y -axis was determined from second harmonic generation data and used to show that this direction also inverts during domain inversion.

Generation of five phase-locked harmonics by implementing a divide-by-three optical frequency divider.

PubMed

Suhaimi, Nurul Sheeda; Ohae, Chiaki; Gavara, Trivikramarao; Nakagawa, Ken'ichi; Hong, Feng-Lei; Katsuragawa, Masayuki

2015-12-15

We report the generation of five phase-locked harmonics, f₁:2403  nm, f₂:1201  nm, f₃:801  nm, f₄:600  nm, and f₅:480  nm with an exact frequency ratio of 1:2:3:4:5 by implementing a divide-by-three optical frequency divider in the high harmonic generation process. All five harmonics are generated coaxially with high phase coherence in time and space, which are applicable for various practical uses.

Thermal effects in high-power CW second harmonic generation in Mg-doped stoichiometric lithium tantalate.

PubMed

Tovstonog, Sergey V; Kurimura, Sunao; Suzuki, Ikue; Takeno, Kohei; Moriwaki, Shigenori; Ohmae, Noriaki; Mio, Norikatsu; Katagai, Toshio

2008-07-21

We investigated thermal behaviors of single-pass second-harmonic generation of continuous wave green radiation with high efficiency by quasi-phase matching in periodically poled Mg-doped stoichiometric lithium tantalate (PPMgSLT). Heat generation turned out to be directly related to the green light absorption in the material. Strong relation between an upper limit of the second harmonic power and confocal parameter was found. Single-pass second-harmonic generation of 16.1 W green power was achieved with 17.6% efficiency in Mg:SLT at room temperature.

Feasibility study of generating ultra-high harmonic radiation with a single stage echo-enabled harmonic generation scheme

NASA Astrophysics Data System (ADS)

Zhou, Kaishang; Feng, Chao; Wang, Dong

2016-10-01

The echo enabled harmonic generation (EEHG) scheme holds the ability for the generation of fully coherent soft x-ray free-electron laser (FEL) pulses directly from external UV seeding sources. In this paper, we study the feasibility of using a single stage EEHG to generate coherent radiation in the "water window" and beyond. Using the high-order operating modes of the EEHG scheme, intensive numerical simulations have been performed considering various three-dimensional effects. The simulation results demonstrated that coherent soft x-ray radiation at 150th harmonic (1.77 nm) of the seed can be produced by a single stage EEHG. The decreasing of the final bunching factor at the desired harmonic caused by intra beam scattering (IBS) effect has also been analyzed.

Symmetry in circularly polarized molecular high-order harmonic generation with intense bicircular laser pulses

NASA Astrophysics Data System (ADS)

Yuan, Kai-Jun; Bandrauk, André D.

2018-02-01

We present symmetry effects of laser fields and molecular geometries in circularly polarized high-order harmonic generation by bichromatic counter-rotating circularly polarized laser pulses. Simulations are performed on oriented molecules by numerically solving time-dependent Schrödinger equations. We discuss how electron recollision trajectories by the orthogonal laser field polarizations influence the harmonic polarization by using a time-frequency analysis of harmonics. It is found that orientation-dependent asymmetric ionization in linear molecules due to Coulomb potentials gives rise to a dependence of the polarization on the harmonic frequency. Effects of Coriolis forces are also presented on harmonic generation. Electron recollision trajectories illustrate the effects of the relative symmetry of the field and the molecule, thus paving a method for circularly polarized attosecond pulse generation and molecular orbital imaging in more complex systems.

Characterization of atherosclerotic plaques by cross-polarization optical coherence tomography

NASA Astrophysics Data System (ADS)

Gubarkova, Ekaterina V.; Dudenkova, Varvara V.; Feldchtein, Felix I.; Timofeeva, Lidia B.; Kiseleva, Elena B.; Kuznetsov, Sergei S.; Moiseev, Alexander A.; Gelikonov, Gregory V.; Vitkin, Alex I.; Gladkova, Natalia D.

2016-02-01

We combined cross-polarization optical coherence tomography (CP OCT) and non-linear microscopy based on second harmonic generation (SHG) and two-photon-excited fluorescence (2PEF) to assess collagen and elastin fibers in the development of the atherosclerotic plaque (AP). The study shows potential of CP OCT for the assessment of collagen and elastin fibers condition in atherosclerotic arteries. Specifically, the additional information afforded by CP OCT, related to birefringence and cross-scattering properties of arterial tissues, may improve the robustness and accuracy of assessment about the microstructure and composition of the plaque for different stages of atherosclerosis.

Second-order nonlinear optical microscopy of spider silk

NASA Astrophysics Data System (ADS)

Zhao, Yue; Hien, Khuat Thi Thu; Mizutani, Goro; Rutt, Harvey N.

2017-06-01

Asymmetric β-sheet protein structures in spider silk should induce nonlinear optical interaction such as second harmonic generation (SHG) which is experimentally observed for a radial line and dragline spider silk using an imaging femtosecond laser SHG microscope. By comparing different spider silks, we found that the SHG signal correlates with the existence of the protein β-sheets. Measurements of the polarization dependence of SHG from the dragline indicated that the β-sheet has a nonlinear response depending on the direction of the incident electric field. We propose a model of what orientation the β-sheet takes in spider silk.

Hypericin-mediated selective photomodification of connective tissues

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hovhannisyan, V., E-mail: hovv@phys.ntu.edu.tw; Guo, H. W.; Chen, Y. F., E-mail: yfchen@phys.ntu.edu.tw

2014-12-29

Controllable modification of biological molecules and supramolecular components of connective tissue are important for biophysical and biomedical applications. Through the use of second harmonic generation imaging, two-photon fluorescence microscopy, and spectrofluorimetry, we found that hypericin, a natural pigment, induces photosensitized destruction of collagen fibers but does not affect elastic fibers and lipids in chicken tendon, skin, and blood vessels. We demonstrated the dynamics and efficiency of collagen photomodification and investigated mechanisms of this processes. Our results suggest that hypericin–mediated photoprocesses in biological tissues may be useful in biomedical applications that require selective modification of connective tissues.

A new ultrasonic transducer for improved contrast nonlinear imaging

NASA Astrophysics Data System (ADS)

Bouakaz, Ayache; ten Cate, Folkert; de Jong, Nico

2004-08-01

Second harmonic imaging has provided significant improvement in contrast detection over fundamental imaging. This improvement is a result of a higher contrast-to-tissue ratio (CTR) achievable at the second harmonic frequency. Nevertheless, the differentiation between contrast and tissue at the second harmonic frequency is still in many situations cumbersome and contrast detection remains nowadays as one of the main challenges, especially in the capillaries. The reduced CTR is mainly caused by the generation of second harmonic energy from nonlinear propagation effects in tissue, which hence obscures the echoes from contrast bubbles. In a previous study, we demonstrated theoretically that the CTR increases with the harmonic number. Therefore the purpose of our study was to increase the CTR by selectively looking to the higher harmonic frequencies. In order to be able to receive these high frequency components (third up to the fifth harmonic), a new ultrasonic phased array transducer has been constructed. The main advantage of the new design is its wide frequency bandwidth. The new array transducer contains two different types of elements arranged in an interleaved pattern (odd and even elements). This design enables separate transmission and reception modes. The odd elements operate at 2.8 MHz and 80% bandwidth, whereas the even elements have a centre frequency of 900 kHz with a bandwidth of 50%. The probe is connected to a Vivid 5 system (GE-Vingmed) and proper software is developed for driving. The total bandwidth of such a transducer is estimated to be more than 150% which enables higher harmonic imaging at an adequate sensitivity and signal to noise ratio compared to standard medical array transducers. We describe in this paper the design and fabrication of the array transducer. Moreover its acoustic properties are measured and its performances for nonlinear contrast imaging are evaluated in vitro and in vivo. The preliminary results demonstrate the advantages of such a transducer design for improved contrast detection.

Two-harmonic complex spectral-domain optical coherence tomography using achromatic sinusoidal phase modulation

NASA Astrophysics Data System (ADS)

Lu, Sheng-Hua; Huang, Siang-Ru; Chou, Che-Chung

2018-03-01

We resolve the complex conjugate ambiguity in spectral-domain optical coherence tomography (SD-OCT) by using achromatic two-harmonic method. Unlike previous researches, the optical phase of the fiber interferometer is modulated by an achromatic phase shifter based on an optical delay line. The achromatic phase modulation leads to a wavelength-independent scaling coefficient for the two harmonics. Dividing the mean absolute value of the first harmonic by that of the second harmonic in a B-scan interferogram directly gives the scaling coefficient. It greatly simplifies the determination of the magnitude ratio between the two harmonics without the need of third harmonic and cumbersome iterative calculations. The inverse fast Fourier transform of the complex-valued interferogram constructed with the scaling coefficient, first and second harmonics yields a full-range OCT image. Experimental results confirm the effectiveness of the proposed achromatic two-harmonic technique for suppressing the mirror artifacts in SD-OCT images.

Neurosurgical confocal endomicroscopy: A review of contrast agents, confocal systems, and future imaging modalities

PubMed Central

Zehri, Aqib H.; Ramey, Wyatt; Georges, Joseph F.; Mooney, Michael A.; Martirosyan, Nikolay L.; Preul, Mark C.; Nakaji, Peter

2014-01-01

Background: The clinical application of fluorescent contrast agents (fluorescein, indocyanine green, and aminolevulinic acid) with intraoperative microscopy has led to advances in intraoperative brain tumor imaging. Their properties, mechanism of action, history of use, and safety are analyzed in this report along with a review of current laser scanning confocal endomicroscopy systems. Additional imaging modalities with potential neurosurgical utility are also analyzed. Methods: A comprehensive literature search was performed utilizing PubMed and key words: In vivo confocal microscopy, confocal endomicroscopy, fluorescence imaging, in vivo diagnostics/neoplasm, in vivo molecular imaging, and optical imaging. Articles were reviewed that discussed clinically available fluorophores in neurosurgery, confocal endomicroscopy instrumentation, confocal microscopy systems, and intraoperative cancer diagnostics. Results: Current clinically available fluorescent contrast agents have specific properties that provide microscopic delineation of tumors when imaged with laser scanning confocal endomicroscopes. Other imaging modalities such as coherent anti-Stokes Raman scattering (CARS) microscopy, confocal reflectance microscopy, fluorescent lifetime imaging (FLIM), two-photon microscopy, and second harmonic generation may also have potential in neurosurgical applications. Conclusion: In addition to guiding tumor resection, intraoperative fluorescence and microscopy have the potential to facilitate tumor identification and complement frozen section analysis during surgery by providing real-time histological assessment. Further research, including clinical trials, is necessary to test the efficacy of fluorescent contrast agents and optical imaging instrumentation in order to establish their role in neurosurgery. PMID:24872922