Near-Infrared Spectroscopy Using a Supercontinuum Laser: Application to Long Wavelength Transmission Spectra of Barley Endosperm and Oil.

The supercontinuum laser is a new type of light source, which combines the collimation and intensity of a laser with the broad spectral region of a lamp. Using such a source therefore makes it possible to focus the light onto small sample areas without losing intensity and thus facilitate either rapid or high-intensity measurements. Single seed transmission analysis in the long wavelength (LW) near-infrared (NIR) region is one area that might benefit from a brighter light source such as the supercontinuum laser. This study is aimed at building an experimental spectrometer consisting of a supercontinuum laser source and a dispersive monochromator in order to investigate its capability to measure the barley endosperm using transmission experiments in the LW NIR region. So far, barley and wheat seeds have only been studied using NIR transmission in the short wavelength region up to 1100 nm. However, the region in the range of 2260-2380 nm has previously shown to be particularly useful in differentiating barley phenotypes using NIR spectroscopy in reflectance mode. In the present study, 350 seeds (consisting of 70 seeds from each of five barley genotypes) in 1 mm slices were measured by NIR transmission in the range of 2235-2381 nm and oils from the same five barley genotypes were measured in a cuvette with a 1 mm path length in the range of 2003-2497 nm. The spectra of the barley seeds could be classified according to genotypes by principal component analysis; and spectral covariances with reference analysis of moisture, ß-glucan, starch, protein and lipid were established. The spectral variations of the barley oils were compared to the fatty acid compositions as measured using gas chromotography-mass spectrometry (GC-MS).

Pushing the limit: investigation of hydrodynamic forces on a trapped particle kicked by a laser pulse.

We introduce a new optical technique where a train of short optical pulses is utilized to disturb a trapped microscopic particle. Using fast (250 kHz) and accurate (nm) detection of the position of the particle, accurately synchronized to the repetition rate of the laser pulses, we can coherently superimpose the displacement caused by each individual laser pulse. Thereby we are able to both bypass the influence from the Brownian motion of the trapped particle and to simultaneously increase the ability to localize its average trajectory by √n, where n is the number of repetitive pulses. In the results presented here we utilize a train of 1200 pulses to kick a 5 µm polystyrene sphere and obtain a spatial resolution corresponding to 0.09 nm and a time resolution of 4 µs. The magnitude of the optical force pushing the particle corresponds to ∼ 10(4)g and enables an investigation of both the hydrodynamical drag and the inertial effects caused by the particle and the surrounding liquid. Our results enables a more accurate testing of the existing extended models for the hydrodynamic drag and we discuss the observed agreement between experiments and theory.

Spectroscopy and picosecond dynamics of aqueous NO2.

We investigate the formation of aqueous nitrogen dioxide, NO2 formed through femtosecond photolysis of nitrate, NO3⁻ and nitromethane CH3NO2(aq). Common to the experiments is the observation of a strong induced absorption at 1610 ± 10 cm(-1), assigned to the asymmetric stretch vibration in the ground state of NO2. This assignment is substantiated through isotope experiments substituting (14)N by (15)N, experiments at different pH values, and by theoretical calculations and simulations of NO2-D2O clusters.

Generation of infrared supercontinuum radiation: spatial mode dispersion and higher-order mode propagation in ZBLAN step-index fibers.

Using femtosecond upconversion we investigate the time and wavelength structure of infrared supercontinuum generation. It is shown that radiation is scattered into higher order spatial modes (HOMs) when generating a supercontinuum using fibers that are not single-moded, such as a step-index ZBLAN fiber. As a consequence of intermodal scattering and the difference in group velocity for the modes, the supercontinuum splits up spatially and temporally. Experimental results indicate that a significant part of the radiation propagates in HOMs. Conventional simulations of super-continuum generation do not include scattering into HOMs, and including this provides an extra degree of freedom for tailoring supercontinuum sources.

IR microscopy utilizing intense supercontinuum light source.

Combining the molecular specificity of the infrared spectral region with high resolution microscopy has been pursued by researchers for decades. Here we demonstrate infrared supercontinuum radiated from an optical fiber as a promising new light source for infrared microspectroscopy. The supercontinuum light source has a high brightness and spans the infrared region from 1400 nm to 4000 nm. This combination allows contact free high resolution hyper spectral infrared microscopy. The microscope is demonstrated by imaging an oil/water sample with 20 µm resolution.

Motion analysis of optically trapped particles and cells using 2D Fourier analysis.

Motion analysis of optically trapped objects is demonstrated using a simple 2D Fourier transform technique. The displacements of trapped objects are determined directly from the phase shift between the Fourier transform of subsequent images. Using end- and side-view imaging, the stiffness of the trap is determined in three dimensions. The Fourier transform method is simple to implement and applicable in cases where the trapped object changes shape or where the lighting conditions change. This is illustrated by tracking a fluorescent particle and a myoblast cell, with subsequent determination of diffusion coefficients and the trapping forces.

Vibrational dynamics of deoxyguanosine 5'-monophosphate following UV excitation.

The relaxation dynamics of the DNA nucleotide deoxyguanosine 5'-monophosphate (dGMP) following 266 nm photoexcitation has been studied by transient IR spectroscopy with femtosecond time resolution. The induced dynamics of the amide I (carbonyl) stretch, the asymmetric guanine ring stretch and the phosphate asymmetric stretch are monitored in the region 1000-1800 cm(-1). Excitation and subsequent rapid internal conversion to a "hot" ground state is reflected by depletion of the vibrational ground states of the amide I stretch and guanine ring stretch. However, the vibrational ground state of the phosphate is left unperturbed, indicating the absence of vibrational coupling between the guanine ring system and the phosphate group. The vibrational ground state of the amide I is repopulated in 2.5 ps (±0.2 ps) while it takes 3.7 ps (±0.5 ps) to repopulate the guanine ring vibration. This article discusses two possible relaxation pathways of dGMP, as well as the implications of the weak phosphate dynamics.

Thermodynamic investigations of methyl tert-butyl ether and water mixtures.

Interactions between methyl tert-butyl ether (MTBE) and water have been investigated by scanning calorimetry, isothermal titration calorimetry, densitometry, IR-spectroscopy, and gas chromatography. The solubilization of MTBE in water at 25 °C at infinite dilution has ΔH° = -17.0 ± 0.6 kJ mol(-1); ΔS° = -80 ± 2 J mol(-1) K(-1); ΔC(p) = +332 ± 15 J mol(-1) K(-1); ΔV° = -18 ± 2 cm(3) mol(-1). The signs of these thermodynamic functions are consistent with hydrophobic interactions. The occurrence of hydrophobic interaction is further substantiated as IR absorption spectra of MTBE-water mixtures show that MTBE strengthens the hydrogen bond network of water. Solubilization of MTBE in water is exothermic whereas solubilization of water in MTBE is endothermic with ΔH° = +5.3 ± 0.6 kJ mol(-1). The negative mixing volume is explained by a large negative contribution due to size differences between water and MTBE and by a positive contribution due to changes in the water structure around MTBE. Henry's law constants, K(H), were determined from vapor pressure measurements of mixtures equilibrated at different temperatures. A van't Hoff analysis of K(H) gave ΔH(H)° = 50 ± 1 kJ mol(-1) and ΔS(H)° = 166 ± 5 J mol(-1) K(-1) for the solution to gas transfer. MTBE is excluded from the ice phase water upon freezing MTBE-water mixtures.

The hunt for HCO(aq).

In contrast to its strong presence in gas-phase reactions, the formyl radical, HCO, has never been identified in aqueous solution. Here the photolysis of aqueous formate anions, HCOO(-)(aq), following the excitation of the (n pi*) transition at 200 nm is studied by infrared femtosecond transient absorption spectroscopy with the purpose of identifying the aqueous formyl radical photoproduct, HCO(aq). However, HCO(aq) is not observed. The experiments indicate that HCO(aq) exists for less than one picosecond before it reacts with the surrounding water molecules.

Femtosecond photolysis of aqueous formamide.

In this work, we investigate the primary photodynamics of aqueous formamide. The formamide was photolyzed using 200 nm femtosecond pulses, and formation of products and their relaxation was followed with approximately 300 fs time resolution using probe pulses covering the range from 193 to 700 nm. Following excitation, the majority of formamide molecules (approximately 80%) converts the electronic excitation energy to vibrational excitation, which effectively is dissipated to the solvent through vibrational relaxation in just a few picoseconds. The vibrational relaxation is observed as a distinct modulation of the electronic absorption spectrum of formamide. The relaxation process is modeled by a simple one-dimensional wavepacket calculation. A smaller fraction of the excited formamide molecules dissociates to the CHO and NH2 radical pairs, of which 50% escape recombination. In addition to the electronic excitation of formamide, we also observe a small contribution from one-photon ionization of formamide and two-photon ionization and dissociation of the water solvent.

All-fiber mode-locked fiber laser.

An environmentally stable mode-locked fiber laser based on nonlinear polarization rotation is experimentally demonstrated. The laser is based on a novel laser configuration that has negligible low-power steady-state reflectivity from one side and, consequently, no CW gain. The laser is self starting and the configuration is implementable as an all-fiber laser with standard polarization-maintaining fiber-pigtailed components. A pulse duration of 5.6 ps is obtained at a repetition rate of 5.96 MHz and at an average power of 8 mW. As an application of the proposed laser configuration, 213 mW of supercontinuum (600-1750 nm) was demonstrated from a fiber laser system with no sections of free-space optics.

Tunable light source for coherent anti-Stokes Raman scattering microspectroscopy based on the soliton self-frequency shift.

We present a photonic crystal fiber (PCF)-based light source for generating tunable excitation pulses (pump and Stokes) that are applicable to coherent anti-Stokes Raman scattering (CARS) microspectroscopy. The laser employed is an unamplified Ti:sapphire femtosecond laser oscillator. The CARS pump pulse is generated by spectral compression of a laser pulse in a PCF. The Stokes pulse is generated by redshifting a laser pulse in a PCF through the soliton self-frequency shift. This setup allows for probing up to 4000 cm(-1) with a spectral resolution of approximately 25 cm(-1). We characterize the stability and robustness of CARS microspectroscopy employing this light source.

Investigation of the primary photodynamics of the aqueous formate anion.

We have investigated the primary photodynamics of the aqueous formate anion using femtosecond transient absorption spectroscopy. The formate anions are excited at 200 nm, and the resulting products are probed in the region 200-650 nm. The ultraviolet part of the transient spectrum compares favorably with that of O-(aq). However, its counter radical, HCO(aq), is not observed. In the visible region hydrated electrons are observed. The electrons are produced from photodetachment of the formate anions and from two-photon ionization of water.

An active interferometer-stabilization scheme with linear phase control.

We report a simple and robust computer-based active interferometer stabilization scheme which does not require modulation of the interfering beams and relies on an error signal which is linearly related to the optical path difference. In this setup, a non-collinearly propagating reference laser beam stabilizes the interference output of the laser light propagating collinearly through the interferometer. This stabilization scheme enables adjustable phase control with 20 ms switching times in the range from 0.02pi radians to 6pi radians at 632.8 nm.

Picosecond anti-Stokes generation in a photonic-crystal fiber for interferometric CARS microscopy.

We generate tunable picosecond anti-Stokes pulses by four-wave mixing of two picosecond pump and Stokes pulse trains in a photonic-crystal fiber. The visible, spectrally narrow anti-Stokes pulses with shifts over 150 nm are generated without generating other spectral features. As a demonstration, we employ the generated anti-Stokes pulses as reference pulses in an interferometric coherent anti-Stokes Raman scattering imaging experiment showing that interpulse coherence among the pump, Stokes and anti-Stokes beams is retained.

Spectral compression of femtosecond pulses in photonic crystal fibers.

We demonstrate efficient spectral compression of femtosecond pulses near the zero-dispersion wavelength in nonlinear photonic crystal fibers (PCFs). The highest measured compression factor is 21, in which case the spectral brightness increases by a factor of 5. We numerically model the pulse propagation and find good agreement with the experiment. We argue that the fibers studied allow for spectral narrowing of more than 2 orders of magnitude. With dispersion-shifted PCFs, efficient spectral compression can take place across the visible and near-infrared part of the spectrum.

Plastic particles at the LASIK interface.

PURPOSE: To investigate the origin, composition, and persistence of the interface particles that frequently are observed after LASIK. DESIGN: Small case series and experimental animal study. METHODS: Four patients received LASIK using a Schwind Supratome (Schwind, Kleinostheim, Germany) and a MEL 70 G-Scan excimer laser (Asclepion, Jena, Germany) and were examined over the course of 1 year using slit-lamp and in vivo confocal microscopy. Four rabbits received a monocular microkeratome incision and were examined immediately after surgery without lifting the flap. After monthly evaluation for 4 months using in vivo confocal microscopy, 2 corneas were processed for histologic analysis and were sectioned serially. To measure the iron content, atomic absorption spectrometry was performed on 2 operated and 2 unoperated rabbit corneas. The chemical composition of the metal and plastic parts of the microkeratome blade was identified using energy dispersive x-ray fluorescence (metal part), and Raman and infrared spectroscopy (plastic part). Before and after oscillation in air, the microkeratome blade and motor-head were examined using light and fluorescence microscopy. In serial sections, interface particles were identified by fluorescence microscopy and their chemical composition was determined using Coherent Antistokes Raman Scattering microscopy. RESULTS: In LASIK patients, thousands of brightly reflecting particles (up to 30 micro m) were observed throughout the interface. The highest particle density was detected where the microkeratome blade had first entered the cornea. Both in the center and at the flap edge, the morphologic features, distribution, and density of these particles remained unaltered throughout the 1-year observation period. In rabbit corneas, interface particles were observed immediately after the microkeratome incision, even though the flap had not been lifted. These particles were similar to those observed in humans and persisted unaltered throughout the study. The operated and unoperated rabbit corneas had comparable iron content, demonstrating that the particles were not fragments of the uncoated steel blade. Only a few particles were observed on the unused microkeratome motor head and blade, whereas numerous fluorescent particles were detected after oscillation in air, the amount of particles increasing with oscillation time. Interestingly, the only fluorescent part of the microkeratome was the plastic segment of the blade. This plastic (polyetherimide) emitted fluorescence identical to that of the observed particles, whereas all metal parts of the microkeratome blade and motor head were nonfluorescent. In serial sections, interface particles showed fluorescent properties equivalent to polyetherimide and exhibited molecular resonance at 1780 and 3100 cm(-1), in accordance with the Raman spectrum of polyetherimide. CONCLUSIONS: Numerous plastic particles are generated during microkeratome oscillation and are deposited at the interface during LASIK. The particles persist unaltered for at least 1 year.

Coherent anti-Stokes Raman scattering microscopy with a photonic crystal fiber based light source.

A coherent anti-Stokes Raman scattering microscope based on a Ti:sapphire femtosecond oscillator and a photonic crystal fiber is demonstrated. The nonlinear response of the fiber is used to generate the additional wavelength needed in the Raman process. The applicability of the setup is demonstrated by imaging of micrometer-sized polystyrene beads.