MULTIPHOTON MICROSCOPIC IMAGING OF MOUSE INTESTINAL MUCOSA BASED ON TWO-PHOTON EXCITED FLUORESCENCE AND SECOND HARMONIC GENERATION

Multiphoton microscopy(MPM),based on two-photon excited fuorescence and second harmonic generation,enables direct noninvasive visualization of tissue architecture and cell morphology in live tissues without the administration of exogenous contrast agents.In this paper,we used MPM to image the microstructures of the mucosa in fresh,unfixed,and unstained intestinal tissue of mouse.The morphology and distribution of the main components in mucosa layer such as columnar cells,goblet cells,intestinal glands,and a little collagen fibers were clearly observed in MPM images,and then compared with standard H&:E images from paired specimens.Our results indicate that MPM combined with endoscopy and miniaturization probes has the potential application in the clinical diagnosis and in vivo monitoring of early intestinal cancer.

Observations of intracellular second-harmonic generation imaging in black phosphorus nanosheets

Functionalized black phosphorus(BP)nanosheets have been considered as promising nanoagents in cancer therapy due to their excellent photothermal conversion efficiency.However,it is still difficult to visually monitor the dynamic localization of BP nanoagents in cancer cells.In this paper,we systematically studied the second-harmonic generation(SHG)signals originating from exfoliated BP nanosheets.Interestingly,under the excitation of a high frequency pulsed laser at 950 nm,the SHG signals of BP nanosheets in vitro are almost undetectable because of their poor stability.However,the intracellular SHG signals from BP nanosheets could be measured by in vivo optical imaging due to the efficient enrichment of living HeLa cells.Moreover,the SHG signal intensity from BP nanosheets increases with the prolonged incubation time.It can be expected that the BP nanosheets could be a promising intracellular SHG nanoprobe employed for visually in vivo biomedical imaging in practical cancer photothermal therapy(PIT).

Evaluation of quantitative contrast harmonic imaging to assess malignancy of liver tumors:A prospective controlled two-center study

AIM: To establish the extent to which contrast enhancement with SonoVue in combination with quantitative evaluation of contrast-medium dynamics facilitates the detection of hepatic tumors. METHODS: One hundred patients with histologically confirmed malignant or benign hepatic tumor (maximum size 5 cm) were analyzed. Contrast-enhanced ultrasound (bolus injection 2.5 mL SonoVue) was carried out with intermittent breath-holding technique using a multifrequency transducer (2.5-4 MHz). Native vascularization was analyzed with power Doppler. The contrast-enhanced dynamic ultrasound investigation was carried out with contrast harmonic imaging in true detection mode during the arterial,portal venous and late phases. Mechanical index was set at 0.15. Perfusion analysis was performed by post-processing of the raw data time intensity curve (TIC) analysis. The cut-off of the gray value differences between tumor and normal liver tissue was established using Receiver Operating Characteristic (ROC) analysis 64-line multi-slice computed tomography served as reference method in all cases. Magnetic resonance tomography was used additionally in 19 cases. RESULTS: One hundred patients with 59 malignant (43 colon,5 breast,2 endocrine metastases,7 hepatocellular carcinomas and 2 kidney cancers) and 41 benign (15 hemangiomas,7 focal nodular hyperplasias,5 complicated cysts,2 abscesses and 12 circumscribed fatty changes) tumors were included. The late venous phase proved to be the most sensitive for classification of the tumor type. Fifty-eight of the 59 malignant tumors were classified as true positive,and one as false negative. This resulted in a sensitivity of 98.3%. Of the 41 benign tumors,37 were classified as true negative and 4 as false negative,which corresponds to a specificity of 90.2%. Altogether,95.0% of the diagnoses were classified as correct on the basis of the histological classification. No investigator-dependency (P = 0.23) was noted. CONCLUSION: The results show the possibility of accurate prediction of malignancy of hepatic tumors with a positive prognostic value of 93.5% using advanced contrast-enhanced ultrasound. Contrast enhancement with SonoVue in combination with quantitative evaluation of contrast-medium dynamics is a valuable tool to discriminate hepatic tumors.

Contrast-enhanced harmonic ultrasound imaging in ablation therapy for primary hepatocellular carcinoma

The success rate of percutaneous radiofrequency(RF)ablation for hepatocellular carcinoma(HCC)depends on correct targeting via an imaging technique.However,RF electrode insertion is not completely accurate for residual HCC nodules because B-mode ultrasound(US),color Doppler,and power Doppler US findings cannot adequately differentiate between treated and viable residual tumor tissue.Electrode insertion is also difficult when we must identify the true HCC nodule among many large regenerated nodules in cirrhotic liver.Two breakthroughs in the field of US technology,harmonic imaging and the development of second-generation contrast agents,have recently been described and have demonstrated the potential to dramatically broaden the scope of US diagnosis of hepatic lesions.Contrast-enhanced harmonic US imaging with an intravenous contrast agent can evaluate small hypervascular HCC even when B-mode US cannot adequately characterize tumor.Therefore,contrast-enhanced harmonic US can facilitate RF ablation electrode placement in hypervascular HCC,which is poorly depicted by B-mode US.The use of contrast-enhanced harmonic US in ablation therapy for liver cancer is an efficient approach.

Cumulative second-harmonic generation of Lamb waves propagating in a two-layered solid plate

The physical process of cumulative second-harmonic generation of Lamb waves propagating in a two-layered solid plate is presented by using the second-order perturbation and the technique of nonlinear reflection of acoustic waves at an interface. In general, the cumulative second-harmonic generation of a dispersive guided wave propagation does not occur. However, the present paper shows that the second-harmonic of Lamb wave propagation arising from the nonlinear interaction of the partial bulk acoustic waves and the restriction of the three boundaries of the solid plates does have a cumulative growth effect if some conditions are satisfied. Through boundary condition and initial condition of excitation, the analytical expression of cumulative second-harmonic of Lamb waves propagation is determined. Numerical results show the cumulative effect of Lamb waves on second-harmonic field patterns.

Second Harmonic Generation of Lamb Wave in Numerical Perspective

The influences of phase and group velocity matching on cumulative second harmonic generation of Lamb waves are investigated in numerical perspective. Finite element simulations of nonlinear Lamb wave propagation are performed for Lamb wave mode pairs with exact and approximate phase velocity matching, with and without group velocity matching, respectively. The evolution of time-domain second harmonic Lamb waves is analyzed with the propagation distance. The amplitudes of primary and second harmonic waves are calculated to characterize the acoustic nonlinearity. The results verify that phase velocity matching is necessary for generation of the cumulative second harmonic Lamb wave in numerical perspective, while group velocity matching is demonstrated to not be a necessary condition.

A 0.33-THz second-harmonic frequency-tunable gyrotron

Dynamics of the axial mode transition process in a 0.33-THz second-harmonic gyrotron is investigated to reveal the physical mechanism of realizing broadband frequency tuning in an open cavity circuit. A new interaction mechanism about propagating waves, featured by wave competition and wave cooperation, is presented and provides a new insight into the beam-wave interaction. The two different features revealed in the two different operation regions of low-order axial modes （LOAMs） and high-order axial modes （HOAMs） respectively determine the characteristic of the overall performance of the device essentially. The device performance is obtained by the simulation based on the time-domain nonlinear theory and shows that using a 12-kV/150-mA electron beam and TE 3,4 mode, the second harmonic gyrotron can generate terahertz radiations with frequency-tuning ranges of about 0.85 GHz and 0.60 GHz via magnetic field and beam voltage tuning, respectively. Additionally, some non-stationary phenomena in the mode startup process are also analyzed. The investiga- tion in this paper presents guidance for future developing high-performance frequency-tunable gyrotrons toward terahertz applications.

Second harmonic magnetoacoustic responses of magnetic nanoparticles in magnetoacoustic tomography with magnetic induction

Due to the unique magnetic, mechanical and thermal properties, magnetic nanoparticles(MNPs) have comprehensive applications as the contrast and therapeutic agents in biomedical imaging and magnetic hyperthermia. The linear and nonlinear magnetoacoustic responses determined by the magnetic properties of MNPs have attracted more and more attention in biomedical engineering. By considering the relaxation time of MNPs, we derive the formulae of second harmonic magnetoacoustic responses(2H-MARs) for a cylindrical MNP solution model based on the mechanical oscillations of MNPs in magnetoacoustic tomography with magnetic induction(MAT-MI). It is proved that only the second harmonic magnetoacoustic oscillations can be generated by MNPs under an alternating magnetic excitation. The acoustic pressure of the 2H-MAR is proportional to the square of the magnetic field intensity and exhibits a linear increase with the concentration of MNPs. Numerical simulations of the 2H-MAR are confirmed by the experimental measurements for various magnetic field intensities and solution concentrations using a laser vibrometer. The favorable results demonstrate the feasibility of the harmonic measurements without the fundamental interference of the electromagnetic excitation, and suggest a new harmonic imaging strategy of MAT-MI for MNPs with enhanced spatial resolution and improved signal-to-noise ratio in biomedical applications.

Temporal electric field of a helium plasma jet by electric field induced second harmonic(E-FISH)method

Electric field is an important parameter of plasma,which is related to electron temperature,electron density,excited species density,and so on.In this work,the electric field of an atmospheric pressure plasma jet is diagnosed by the electric field induced second harmonic(E-FISH)method,and the time-resolved electric field under different conditions is investigated.When positive pulse voltage is applied,the electric field has a peak of about 25 kV cm-1at the rising edge of the voltage pulse.A dark channel is left behind the plasma bullet and the electric field in the dark channel is about 5 kV cm-1.On the other hand,when negative pulse voltage is applied,the electric field has a peak of-16 kV cm-1when the negative voltage is increased to-8 kV.A relatively bright channel is left behind the plasma head and the electric field in this relatively bright channel is about-6 kV cm-1.When the pulse rising time increases from 60 to 200 ns,the peak electric field at both the rising edge and the falling edge of the voltage decreases significantly.When 0.5%of oxygen is added to the main working gas helium,the peak electric field at the rising edge is only about 15 kV cm-1.On the other hand,when 0.5%nitrogen is added,the peak electric field increases especially at the falling edge of the voltage pulse,where it increases reversely from-12 to-16 kV cm-1(the minus sign only represents the direction of electric field).

Photoinduced Second Harmonic Generation of Bi_2S_3 Microcrystallite Doped Silica Glass

Silica glasses doped with Bi2S3 microcystallite was prepared by the sol-gel process. Photoinduced second harmonic generation (SHG) was observed in the glass when it was irradiated with intense 1.06 mum and frequency doubled laser beams from a mode-locked Nd: YAG laser. It was found that the signal intensity increased with the irradiating time and approached a saturation gradually. The effect may be explained reasonably by the DC field model.

Dual bound states in the continuum enhanced second harmonic generation with transition metal dichalcogenides monolayer

The emergence of two dimensional(2D)materials has opened new possibilities for exhibiting second harmonic genera-tion(SHG)at the nanoscale,due to their remarkable optical response related to stable excitons at room temperature.However,the ultimate atomic-scale interaction length with light makes the SHG of Transition Metal Dichalcogenides(TM-Ds)monolayers naturally weak.Here,we propose coupling a monolayer of TMDs with a photonic grating slab that works with doubly resonant bound states in the continuum(BIC).The BIC slabs are designed to exhibit a pair of BICs,reson-ant with both the fundamental wave(FW)and the second harmonic wave(SHW).Firstly,the spatial mode matching can be fulfilled by tilting FW's incident angle.We theoretically demonstrate that this strategy leads to more than four orders of magnitude enhancement of SHG efficiency than a sole monolayer of TMDs,under a pump light intensity of 0.1 GW/cm^(2).Moreover,we demonstrate that patterning the TMDs monolayer can further enhance the spatial overlap coefficient,which leads to an extra three orders of magnitude enhancement of SHG efficiency.These results demonstrate remarkable pos-sibilities for enhancing SHG with nonlinear 2D materials,opening many opportunities for chip-based light sources,nano-lasers,imaging,and biochemical sensing.

Theory of higher harmonics imaging in tapping-mode atomic force microscopy

The periodic impact force induced by tip-sample contact in a tapping mode atomic force microscope （AFM） gives rise to the non-harmonic response of a micro-cantilever. These non-harmonic signals contain the full characteristics of tip-sample interaction. A complete theoretical model describing the dynamical behaviour of tip-sample system was developed in this paper. An analytic formula was introduced to describe the relationship between time-varying tip-sample impact force and tip motion. The theoretical analysis and numerical results both show that the timevarying tip-sample impact force can be reconstructed by recording tip motion. This allows for the reconstruction of the characteristics of the tip-sample force, like contact time and maximum contact force. It can also explain the ability of AFM higher harmonics imaging in mapping stiffness and surface energy variations.

1.5 Harmonic Imaging Sonography with microbubble contrast agent improves characterization of hepatocellular carcinoma

AIM： TO investigate the usefulness of 1.5 Harmonic Imaging Sonography with the use of the contrast agent Levovist for the diagnosis of hepatocellular carcinoma （HCC） and for the evaluation of therapeutic response. METHODS： Phantom experiments were performed to compare the contrast effects of 2^rd harmonic imaging and 1.5 Harmonic Imaging Sonography. 1.5 Harmonic Imaging Sonography was employed to examine 36 patients with HCC （42 nodules） before and after the treatment and to compare against the findings obtained using other diagnostic imaging modaUties. RESULTS： In 1.5 Harmonic Imaging Sonography, the tumor vessels of HCCs were clearly identified during the early phase, and late-phase images clearly demonstrated the differences in contrast enhancement between the tumor and surrounding hepatic parenchyma. Blood flow within the tumor was detected in 36 nodules （85.7%） during the early phase and in all 42 nodules （100%） during the late phase using 1.5 Harmonic Imaging Sonography, in 38 nodules （90.5%） using contrast-enhanced CT, in 34 nodules （81.0%） using digital subtraction angiography （DSA）, and in 42 nodules （100%） using US CO2angiography. Following transcatheter arterial embolization, 1.5 Harmonic Imaging Sonography detected blood flow and contrast enhancement within the tumors that were judged to contain viable tissue in 20 of 42 nodules （47.6%）. However, 6 of these 20 cases were not judged in contrastenhanced CT. 1.5 Harmonic Imaging Sonography was compared with the US CO2 angiography findings as the gold standard, and the sensitivity and specificity of these images for discerning viable and nonviable HCC after transcatheter arterial embolization were 100% and 100%,respectively. CONCLUSION： 1.5 Harmonic Imaging Sonography permits the vascular structures of HCCs to be identified and blood flow within the tumor to be clearly demonstrated. Furthermore, 1.5 Harmonic Imaging Sonography is potentially useful for evaluating the therapeutic effects of transcatheter arterial embolization on HCC. 2005 The WJG Press and Elsevier Inc. All rights reserved

Second Harmonic Generation in Some Donor-Acceptor Substituted Chalcone Derivatives

A series of five chalcone derivatives with different substituents in para and meta posions have been synthesized, and single crystals were successfully grown in aceton solution by slow evaporation solution growth technique (SESGT). Single crystal X-ray studies revealed that all the crystals crystallized in noncentrosymmetric space group with their molecular dipoles perfectly aligned in a direction-favorable for large nonlinear optical effects. Kurtz powder tests revealed that all five materials have second-harmonic-generating properties with maximum efficiencies of approximately 14 times that of urea standard. UV-vis-NIR spectroscopy and thermogravimetric analyses are also presented for all of the reported materials. Among the five chalcones, high quality single crystals of 4-Methoxy-4'-chlorochalcone were grown by SESGT, and its crystalline perection were studied by using a high resolution X-ray diffractometry (HRXRD).

Disorder Effect on the Transmission of Second Harmonic Waves in One-Dimensional Periodically Poled LiTaO₃

One of the methods for calculating electromagnetic wave dispersion in multi-layer structures is the transfer matrix method. In this paper, we use the transfer matrix method for second harmonic generation in a nonlinear multilayer structure. The nonlinear photonic crystals investigated in this paper are as one-dimensional multi-layered structures including ferroelectric materials such as LiTaO3. Our goal is to investigate the effect of the disorder on the transmission spectrum of electromagnetic waves. Our results showed that positional disorder has different effects on the transmitting band and the gap band. The disorder in the transmitting band reduces the transmission coefficient of the waves and increases the transmission coefficient of the waves in the gap band. Such work has not yet been done on nonlinear photonic crystals producing the second harmonic.

Lock-in-Amplifier Model for Analyzing the Behavior of Signal Harmonics in Magnetic Particle Imaging

The purpose of this study was to present a lock-in-amplifier model for analyzing the behavior of signal harmonics in magnetic particle imaging (MPI) and some simulation results based on this model. In the lock-in-amplifier model, the signal induced by magnetic nanoparticles (MNPs) in a receiving coil was multiplied with a reference signal, and was then fed through a low-pass filter to extract the DC component of the signal (output signal). The MPI signal was defined as the mean of the absolute value of the output signal. The magnetization and particle size distribution of MNPs were assumed to obey the Langevin theory of paramagnetism and a log-normal distribution, respectively, and the strength of the selection magnetic field (SMF) in MPI was assumed to be given by the product of the gradient strength of the SMF and the distance from the field-free region (x). In addition, Gaussian noise was added to the signal induced by MNPs using normally-distributed random numbers. The relationships between the MPI signal and x were calculated for the odd- and even-numbered harmonics and were investigated for various time constants of the low-pass filter used in the lock-in amplifier and particle sizes and their distributions of MNPs. We found that the behavior of the MPI signal largely depended on the time constant of the low-pass filter and the particle size of MNPs. This lock-in-amplifier model will be useful for better understanding, optimizing, and developing MPI, and for designing MNPs appropriate for MPI.

Time-domain analysis of second-harmonic generation of primary Lamb wave propagation in an elastic plate

Within the second-order perturbation approximation, this paper investigates the physical process of generation of the time-domain second harmonic by a primary Lamb wave waveform in an elastic plate. The present work is performed based on the preconditions that the phase velocity matching is satisfied and that the transfer of energy from the primary Lamb wave to the double frequency Lamb wave is not zero. It investigates the influences of the difference between the group velocities of the primary Lamb wave and the double frequency Lamb wave, the propagation distance and the duration of the primary Lamb wave waveform on the envelope shape of the time-domain second harmonic. It finds that the maximum magnitude of the envelope of the second-harmonic waveform can grow within some propagation distance even if the condition of group velocity matching is not satisfied. Our analyses also indicate that the maximum magnitude of the envelope of the second-harmonic waveform is kept constant beyond a specific propagation distance. Furthermore, it concludes that the integration amplitude of the time-domain second-harmonic waveform always grows with propagation distance within the second-order perturbation. The present research yields new physical insight not previously available into the effect of generation of the time-domain second harmonic by propagation of a primary Lamb wave waveform.

Enhancement effect of cumulative second-harmonic generation by closed propagation feature of circumferential guided waves

On the basis of second-order perturbation approximate and modal expansion approach,we investigate the enhancement effect of cumulative second-harmonic generation(SHG)of circumferential guided waves(CGWs)in a circular tube,which is inherently induced by the closed propagation feature of CGWs.An appropriate mode pair of primary-and double-frequency CGWs satisfying the phase velocity matching and nonzero energy flux is selected to ensure that the second harmonic generated by primary CGW propagation can accumulate along the circumference.Using a coherent superposition of multi-waves,a model of unidirectional CGW propagation is established for analyzing the enhancement effect of cumulative SHG of primary CGW mode selected.The theoretical analyses and numerical simulations performed directly demonstrate that the second harmonic generated does have a cumulative effect along the circumferential direction and the closed propagation feature of CGWs does enhance the magnitude of cumulative second harmonic generated.Potential applications of the enhancement effect of cumulative SHG of CGWs are considered and discussed.The theoretical analysis and numerical simulation perspective presented here yield an insight previously unavailable into the physical mechanism of the enhancement effect of cumulative SHG by closed propagation feature of CGWs in a circular tube.

Order-Disorder Transition of Carboxyl Terminated Chains in Polydiacetylenes Vesicles Probed by Second Harmonic Generation and Two-Photon Fluorescence

To understand and control the interfacial properties of polydiacetylenes(PDAs)vesicles withπ-conjugated backbone is very important for their colorimetric sensing of chemical and biological targets.In this work,we adopted 10,12-pentacosadiynoic acid(PCDA)as the model molecule to prepare PDAs vesicles in aqueous solution with different forms(from monomer to blue-to-purple-to-red phase)by controlling the UV irradiation dose.The variations of the interfacial conformation of PDAs vesicles during chromatic transitions were inspected by the adsorption behaviors of probe molecules(4-(4-diethylaminostyry)-1-methylpyridinium iodide,D289)on vesicle surface with surface-specific second harmonic generation(SHG)and zeta potential measurements.Resonant SHG signal from D289 adsorbed on vesicle surface attenuated sharply,and the adsorption free energy as well as the corresponding two-photon fluorescence signal decreased slightly in chromatic transitions.While,the change in the surface density of the adsorbed D289 molecules for PDAs vesicles with different forms was relatively small as estimated from zeta potential measurements.The attenuation of the SHG intensity was thus attributed to the overall order-disorder transition and the changed orientation of D289 molecules caused by the gradual distortion of carboxyl head group driven by backbone perturbation.

Surface second harmonic generation of chiral molecules using three-coupled-oscillator model

Based on the three-coupled-oscillator molecular model we proposed, the relation between the second-order susceptibilities of a chiral film and the molecular hyperpolarizabilities is given. The effect of microscopic parameters on the second-order susceptibilities is simulated numerically and the difference between the efficiencies of s-polarized secondharmonic fields induced by the left- and the right-handed circularly-polarized fundamental beams is discussed. The theoretical basis for studying second-order nonlinear optical properties of the chiral molecular media with a tripod-like structure is provided in this paper.