Efficient non-degenerate two-photon excitation for fluorescence microscopy.

Non-degenerate two-photon excitation (ND-TPE) has been explored in two-photon excitation microscopy. However, a systematic study of the efficiency of ND-TPE to guide the selection of fluorophore excitation wavelengths is missing. We measured the relative non-degenerate two-photon absorption cross-section (ND-TPACS) of several commonly used fluorophores (two fluorescent proteins and three small-molecule dyes) and generated 2-dimensional ND-TPACS spectra. We observed that the shape of a ND-TPACS spectrum follows that of the corresponding degenerate two-photon absorption cross-section (D-TPACS) spectrum, but is higher in magnitude. We found that the observed enhancements are higher than theoretical predictions.

Measurement of the relative non-degenerate two-photon absorption cross-section for fluorescence microscopy.

In non-degenerate two-photon microscopy (ND-TPM), the required energy for fluorescence excitation occurs via absorption of two photons of different energies derived from two synchronized pulsed laser beams. ND-TPM is a promising imaging technology offering flexibility in the choice of the photon energy for each beam. However, a formalism to quantify the efficiency of two-photon absorption (TPA) under non-degenerate excitation, relative to the resonant degenerate excitation, is missing. Here, we derive this formalism and experimentally validate our prediction for a common fluorophore, fluorescein. An accurate quantification of non-degenerate TPA is important to optimize the choice of photon energies for each fluorophore.

The strong competitive role of 2n pollen in several polyploidy hybridizations in Rosa hybrida.

BACKGROUND: 2n pollen play a strong competitive role in hybridization and breeding of multiploids in Rosa hybrida. The ploidy inheritable characteristic of 'Orange Fire' × 'Old Blush' were analyzed. RESULT: The results of the cytological observations indicated that 2n pollen developed from the defeated cytoplasmic division or nuclear division in the meiosis metaphase II of PMC (pollen mother cell) in 'Old Blush'. The natural generation rate of the 2n pollen in 'Old Blush' (2x) was about 1.39 in percentage of all male gametes, whereas the tetraploids in the F1 offspring possessed a high rate, i.e., 44.00%. The temporal and spatial characteristics of 'Old Blush' pollen germination on the stigma and growth in pistil of 'Orange Fire' and 'DEE' were observed, and the results suggested that the germination rate of 2n pollen on the stigma was not superior to that of 1n pollen, but that the proportion of 2n pollen increased to 30.90 and 37.20%, respectively, while it traversed the stigma and entered into style. The callose plug in the 2n pollen tube was significantly thinner than that of 1n pollen tube. And each trait involved in our experiment probably is very important for F1 morphological phenotypes. CONCLUSION: We conclude that 2n pollen are involved in hybridization and have a competitive advantage while it traversed the stigma and entered into style. The callose plug in the 2n pollen tube was may have strongly influenced the competitive process in R. hybrida.

Electronic Metamaterials with Tunable Second-order Optical Nonlinearities.

The ability to engineer metamaterials with tunable nonlinear optical properties is crucial for nonlinear optics. Traditionally, metals have been employed to enhance nonlinear optical interactions through field localization. Here, inspired by the electronic properties of materials, we introduce and demonstrate experimentally an asymmetric metal-semiconductor-metal (MSM) metamaterial that exhibits a large and electronically tunable effective second-order optical susceptibility (χ(2)). The induced χ(2) originates from the interaction between the third-order optical susceptibility of the semiconductor (χ(3)) with the engineered internal electric field resulting from the two metals possessing dissimilar work function at its interfaces. We demonstrate a five times larger second-harmonic intensity from the MSM metamaterial, compared to contributions from its constituents with electrically tunable nonlinear coefficient ranging from 2.8 to 15.6 pm/V. Spatial patterning of one of the metals on the semiconductor demonstrates tunable nonlinear diffraction, paving the way for all-optical spatial signal processing with space-invariant and -variant nonlinear impulse response.

Plasmonic enhanced two-photon absorption in silicon photodetectors for optical correlators in the near-infrared.

A high-density array of plasmonic coaxial nanoantennas is used to enhance the two-photon absorption (TPA) process in a conventional silicon photodetector from a mode-locked 76 MHz Ti:sapphire laser over a spectral range from 1340 to 1550 nm. This enhanced TPA was used to generate an interferometric autocorrelation trace of a 150 fs laser pulse. Unlike second-harmonic generation, this technique does not require phase matching or a bulky crystal and can be used on a low-cost integrated silicon platform over a wide range of near-IR wavelengths compatible with modern commercial tunable femtosecond sources.

Observation of second-harmonic generation in silicon nitride waveguides through bulk nonlinearities.

We present experimental results on the observation of a bulk second-order nonlinear susceptibility, derived from both free-space and integrated measurements, in silicon nitride. Phase-matching is achieved through dispersion engineering of the waveguide cross-section, independently revealing multiple components of the nonlinear susceptibility, namely χ(2) yyy = 0.14 ± 0.08 pm/V and χ(2) xxy = 0.30 ± 0.18 pm/V. Additionally, we show how the second-harmonic signal may be tuned through the application of bias voltages across silicon nitride. The material properties measured here are anticipated to allow for the realization of new nanophotonic devices in CMOS-compatible silicon nitride waveguides, adding to their viability for telecommunication, data communication, and optical signal processing applications.

Non-degenerate 2-photon excitation in scattering medium for fluorescence microscopy.

Non-degenerate 2-photon excitation (ND-2PE) of a fluorophore with two laser beams of different photon energies offers an independent degree of freedom in tuning of the photon flux for each beam. This feature takes advantage of the infrared wavelengths used in degenerate 3-photon excitation (D-3PE) microscopy to achieve increased penetration depths, while preserving a relatively high 2-photon excitation cross section in comparison to that of D-3PE. Here, using spatially and temporally aligned Ti:Sapphire laser and optical parametric oscillator beams operating at near infrared (NIR) and short-wavelength infrared (SWIR) optical frequencies, we employ ND-2PE and provide a practical demonstration that a constant fluorophore emission intensity is achievable deeper into a scattering medium using ND-2PE as compared to the commonly used degenerate 2-photon excitation (D-2PE).