Membrane potential detection with second-harmonic generation and two-photon excited fluorescence: A theoretical comparison

We theoretically compare the performance of TPEF and SHG microscopy for membrane potential imaging. We argue that electrochromic TPEF and SHG membrane potential responses are reflections of the same phenomenon, and can be described in a unified manner as resulting from the linear Stark effect. We also show that TPEF and SHG exhibit similar sensitivities in the case of both electrochromic and orientational response mechanisms. Despite their similar sensitivities, SHG nevertheless presents advantages over TPEF for membrane potential imaging because of its remarkable spatial and spectral contrast, and because of its insensitivity to non-radiative excited-state damping mechanisms.     

Use of a single-mode fiber coupler for second-harmonic-generation microscopy

We present a compact second-harmonic-generation (SHG) microscope based on a three-port single-mode fiber coupler. The fiber coupler is used to deliver a near-infrared ultrashort-pulsed laser beam as well as to collect the SHG signal in the visible wavelength range. The SHG microscope exhibits an axial resolution of 1.8 microm, which shows a slight enhancement of the optical sectioning effect compared with that for two-photon excitation at the same excitation wavelength. It is also demonstrated that SHG and two-photon fluorescence images under parallel and perpendicular laser excitation polarization can be simultaneously obtained.     

Enhanced second-harmonic generation from individual metallic nanoapertures

We demonstrate the ability of single-subwavelength-size nanoapertures fabricated in a gold metal thin film to enhance second-harmonic generation (SHG) as compared to a bare metal film. Nonlinear microscopy imaging with polarization resolution is used to quantify the SHG enhancement in circular and triangular nanoaperture shapes. The dependence of the measured SHG enhancement on circular aperture diameters is seen to originate from both phase retardation effects and field enhancements at the nanoaperture edge. Triangular nanoapertures exhibit superior SHG enhancement compared with circular ones, as expected from their noncentrosymmetric shape.     

Second-harmonic generation in graded metallic films

We study the effective second-harmonic generation (SHG) susceptibility in graded metallic films by invoking the local field effects exactly and further numerically demonstrate that graded metallic films can serve as a novel optical material for producing a broad structure in both the linear and the SHG response and an enhancement in the SHG signal.     

Second harmonic generation of propagating collective excitations in Bose－Einstein condensates

We consider a possible second harmonic generation (SHG) of propagating collective excitations in a two-component Bose－Einstein condensate (BEC) with repulsive atom－atom interactions. We show that the phase-matching condition for the SHG can be fulfilled if the wave vectors and frequencies of the excitations are chosen adequately from different dispersion branches. We solve the nonlinear amplitude equations for the SHG derived using a method of multiple-scales and provide SHG solutions similar to those obtained for a SHG in nonlinear optical media. A possible experimental realization of the SHG for the propagating collective modes in a cigar-shaped two-component BEC is also discussed.     

Transmission confocal laser scanning microscopy with a virtual pinhole based on nonlinear detection

We present a transmission-mode confocal laser scanning microscope system based on the use of second-harmonic generation (SHG) for signal detection. Our method exploits the quadratic intensity dependence of SHG to preferentially reveal unscattered signal light and reject out-of-focus scattered background. The SHG crystal acts as a virtual pinhole that remains self-aligned without the need for descanning.     

有机/无机薄膜凝胶动态过程的实时光学二次谐波产生研究

采用实时光学二次谐波产生(SHG)测量方法,以两亲性分子半花菁为掺杂探针分子,对溶胶-凝胶技术制备的玻璃薄膜的凝胶干燥的动态过程进行了研究.结合紫外—可见吸收光谱的测量,发现：因两亲性分子在膜中的自取向性可诱导出二阶光学非线性,溶胶-凝胶技术制备的有机/无机复合膜的凝胶干燥过程可通过薄膜SHG强度的变化半定量地反映.凝胶干燥收缩导致有序掺杂分子数减少,膜片的光学非线性减弱.该过程随薄膜处理温度的升高而急剧缩短;另外,有机分子在膜中的聚集程度随膜片处理温度的增加而很快减少. 关键词：     

Enhanced second-harmonic generation for multiple wavelengths by defect modes in one-dimensional photonic crystals

We present an effective design of aperiodically stacked layers of nonlinear material and air, sandwiched by two truncated photonic crystals, in terms of the simulation annealing method. The constructed structure can achieve multiple-wavelength second-harmonic generation (SHG) at the preassigned wavelengths. We derive a general solution of SHG in 1D inhomogeneous systems and apply it to evaluate the SHG conversion efficiency. Numerical simulations show that the conversion efficiency of SHG can be significantly enhanced when the fundamental wave frequencies are assigned to the designed defect states.     

Discrimination of basal cell carcinoma from normal dermal stroma by quantitative multiphoton imaging

We performed multiphoton fluorescence (MF) and second-harmonic generation (SHG) imaging on human basal cell carcinoma samples. In the dermis, basal cell carcinomas can be identified by masses of autofluorescent cells with relatively large nuclei and marked peripheral palisading. In the normal dermis, SHG from dermal collagen contributes largely to the multiphoton signal. However, within the cancer stroma, SHG signals diminish and are replaced by autofluorescent signals, indicating that normal collagen structures responsible for SHG have been altered. To better delineate the cancer cells and cancer stroma from the normal dermis, a quantitative MF to SHG index is developed. We demonstrate that this index can be used to differentiate cancer cells and adjacent cancer stroma from the normal dermis. Our work shows that MF and SHG imaging can be an alternative for Mohs' surgery in the real-time guidance of the secure removal of basal cell carcinoma.     

Disorder-correlated enhancement of second-harmonic generation in strongly photonic porous gallium phosphide

We report an order of magnitude enhancement of second-harmonic generation (SHG) from porous gallium phosphide relative to SHG in crystalline gallium phosphide. Optical heterodyning measurements of photon free-path length reveal a correlation between SHG enhancement and disorder of the porous material.     

Phase matching considerations in second harmonic generation from tissues: Effects on emission directionality, conversion efficiency and observed morphology

We present a heuristic treatment which relates SHG image intensities, signal directionality, and observed morphology to the physical structure of collagen and cellulose fibrillar tissues. The SHG creation model is based upon relaxed phase matching conditions which account for dispersion, randomness, and axial momentum contributions from the media, and includes a mathematical treatment which relates SHG conversion efficiency to fibril diameter and packing through the inclusion of potential intensity amplification resultant from quasi-phase matching (QPM). A direct consequence of this theory is that SHG in biological tissues is not strictly a coherent process, and that the forward directed SHG has a longer coherence length than the backward component, Through this treatment, we show that the emission directionality and also conversion efficiency do not arise solely from the fibril size but also depend on packing density and order of the inter-fibril structure. We demonstrate these principles in comparing the SHG response in normal and Osteogenesis Imperfecta (OI) skin. We show that the observed directionality and decreased relative intensity in the diseased state is consistent with phase matching conditions arising from the decreased fibril size and more random assembly. We further use this theory to explain the differences in morphology seen in forward and backward collected SHG in fibrillar tissues (e.g., collagenous and cellulosic). Specifically, we attribute segmented appearance to destructive interference between small fibrils separated by less than the coherence length. We suggest the approach based on relaxed phasematching conditions is general in predicting the SHG response in tissues and may be broadly applicable in interpreting the SHG contrast for diagnostic applications.     

Second harmonic generation due to quadrupole interaction in a photonic crystal slab: angle dependence and symmetry of the unit cell

We investigate second harmonic generation (SHG) from a photonic crystal slab consisting of centrosymmetric materials. The SHG signal is observed in the transmission direction when the incident laser excites the quasiwaveguide mode. As the SHG frequency approaches the exciton level, the SHG intensity increases resonantly. When the incident angle is exactly 0, the SHG signal vanishes even if the transmission dip is excited. This fact is readily explained by a quadrupole theory based on the Lorentz oscillator model, where the source of the nonlinearity is the Lorentz force. When the unit cell in the photonic crystal lacks inversion symmetry, the SHG signal is expected even for the normal incidence. It is experimentally demonstrated for a square array of triangular semiconductor slabs.     

Monitoring the thermally induced structural transitions of collagen by use of second-harmonic generation microscopy

The thermal disruption of collagen I in rat tail tendon is investigated with second-harmonic generation (SHG) microscopy. We investigate its effects on SHG images and intensity in the temperature range 25 degrees-60 degrees C. We find that the SHG signal decreases rapidly starting at 45 degrees C. However, SHG imaging reveals that breakage of collagen fibers is not evident until 57 degrees C and worsens with increasing temperature. At 57 degrees C, structures of both molten and fibrous collagen exist, and the disruption of collagen appears to be complete at 60 degrees C. Our results suggest that, in addition to intensity measurement, SHG imaging is necessary for monitoring details of thermally induced changes in collagen structures in biomedical applications.     

Theory of enhanced second-harmonic generation in some artificial materials

We review the recent theoretical investigation on enhanced second-harmonic generation (SHG) in soft nonlinear optical materials based on ferrofluids, graded metallic films, and graded metal-dielectric films of anisotropic particles. The SHG of soft ferrofluid-based nonlinear optical materials possess magnetic-field controllabilities, i.e., magnetic-field-controllable anisotropy, red-shift and enhancement, which are caused to appear by the shift of a resonant plasmon frequency due to the formation of the chains of the coated nanoparticles. Both graded metallic films and graded metal-dielectric films of anisotropic particles can serve as a novel optical material for producing a broad structure in both the linear and SHG response and an enhancement in the SHG signal, due to the local field effects.      

A generalized Ginzburg-Landau approach to second harmonic generation

We develop a generalized Ginzburg-Landau theory for second harmonic generation (SHG) in magnets by expanding the free energy in terms of the order parameter in the magnetic phase and the susceptibility tensor in the corresponding high-temperature phase. The non-zero components of the SHG susceptibility in the ordered phase are derived from the symmetries of the susceptibility tensor in the high-temperature phase and the symmetry of the order parameter. In this derivation, the dependence of the SHG susceptibility on the order parameter follows naturally, and therefore its nonreciprocal optical properties. We examine this phenomenology for the magnetoelectric compound Cr₂O₃ as well as for the ferroelectromagnet YMnO₃. Received 27 August 1999     

Collinear second harmonic generation of 20 wavelengths in a single two-dimensional decagonal nonlinear photonic quasi-crystal

We report on the observation of quasi-phase matched collinear second harmonic generation (SHG) at 20 wavelengths in a two-dimensional nonlinear photonic quasi-crystal with decagonal lattice. We show that at some wavelengths the second harmonics are generated via standard quasi-phase matching, namely a reciprocal vector exists that equals the phase-mismatch vector, while at others the SHG is as a result of the projection-based quasi-phase matching in which the momentum conservation is satisfied up to a projection of a reciprocal vector onto the direction of propagation. In spite of different generation mechanisms, the reciprocal vectors (or their projections) involved in the collinear QPM SHG can be described by a generalized equation.     

Implementation and evaluation of a detector for forward propagated second harmonic signals

Second harmonic generation (SHG) is emerging as an alternative non-linear imaging method. The fact that most commercial multi-photon microscopes can be easily adapted to image SHG makes it appealing to explore the kind of sample information given by SHG. Here we describe an SHG detection implementation designed to optimize the collection of forward propagating light. A Hamamatsu H957-08 PMT is inserted at the back-focal plane of the condenser on an inverted Nikon TE300, and controlled by the existing electronics of a BioRad 1024MP system. Evaluation of the performance was done on common SHG generating preparations, KH₂PO₄ crystals and collagen. We concluded that positioning a detector at the back focal plane of the condenser provides a highly efficient detection system for second harmonic signals, with many advantages over a detector sited at the lamp housing.     

Manifestation of hexagonal-like structure in the second-order nonlinear effects in the GaN nanocrystallites

Contribution of the hexagonal-like structural components to the photoinduced second harmonic generation (SHG) in GaN large-sized nanocrystallites (with sizes about the 10–30 nm) incorporated into the polyvinylalcohol photopolymer matrices is revealed. The SHG measurements were done using pulsed Nd:YAG laser beam (λ=1.06 μm; pulse duration τ=15–50 ps, laser power about 30 MW) as a fundamental ones and a picosecond nitrogen pulsed laser (P=10 MW; λ=0.377 μm; pulse time duration τ=10–25 ps) as a photoinducing one. We have found that with increasing pumping power density the SHG output signal increases and achieves its maximum value for the power density about 2.6 GW/cm² per pulse. The maximal output photoinduced SHG signal was achieved for parallel directions of the pumping and fundamental beam polarizations. The maximal values of the second-order nonlinear susceptibilities were equal to about 1.09 pm/V. We have observed an increase of the output SHG below 30 K and for pump-probe delaying time about 18 ps. Substantial contribution to the SHG of wurtzite-like (hexagonal) structural fragments is shown.     

Waveguide second-harmonic generation device with broadened flat quasi-phase-matching response by use of a grating structure with located phase shifts

We report on a theoretical analysis and experiments for bandwidth broadening in quasi-phase-matched (QPM) second-harmonic generation (SHG).We used phase-shifted segments of a periodic grating to obtain a spectrally broadened, nearly flat response simultaneously with high conversion efficiency. We used an x-cut MgO:LiNbO(3) QPM waveguide in our analysis and experiments. The spectral range of the 850-nm fundamental for which SHG conversion exceeded 0.95 of the maximum value broadened from 0.02 to 0.12 nm when a 1-cm-long grating was divided into three segments with optimum phase shift. SHG conversion efficiency was 300%/W for this waveguide. The SHG efficiency and phase-matching characteristics showed good agreement with theoretical results.     

Magnetization-induced anisotropy of second harmonic generation in thin cobalt films

Magnetization-induced effects in the symmetry properties of optical second harmonic generation (SHG) are studied in thin cobalt films. We demonstrate that the application of an external magnetic field leads to the appearance of a strong SHG azimuthal anisotropy from the isotropic Au/Co structure. Symmetry analysis of the SHG dependencies, supported by the SHG interferometry measurements, indicates that the effective magnetic and crystallographic second-order susceptibilities are both of the same order of magnitude.