Application of isotopically labeled methylmercury for isotope dilution analysis of biological samples using gas chromatography/ICPMS

An isotope dilution (ID) procedure for the determination of methylmercury (MMHg) in biological samples using an inductively coupled plasma mass spectrometer as detector after the capillary gas chromatographic separation (CGC/ICPMS) has been developed. For the first time, open-focused-microwave pretreatment has been used in conjunction with ID. Optimum conditions for the measurement of isotope ratios on the fast transient chromatographic peaks have been established. Mass bias was found to be about 1.5%/mass unit and was corrected by using the simultaneously measured thallium signals at 203Tl and 205Tl. After mass-bias correction, deviation of the theoretical mercury ratio values was found to be as low as 0.2%. Isotope ratio precisions based on the peak areas measurements were 0.3% RSD for 20 pg injected (as Hg absolute). The absolute detection limits were in the range of 20-30 fg for 202Hg and 201Hg. Methylmercury enriched in 201Hg has been synthesized by direct reaction with methylcobalamine. The concentration of the MMHg spike has been measured by reverse isotope dilution with a natural MMHg standard. The capabilities of CGC/ICPMS to measure isotope ratios were used to optimize sample derivatization by aqueous ethylation with NaBEt4 with respect to MMHg degradation pathways and quantitative recovery. The accuracy of the method developed has been validated with biological certified reference materials (CRM-463, DORM-1).     

An electrospray ionization source for integration with microfluidics

We have demonstrated a new electrospray ionization (ESI) device incorporating a tip made from a shaped thin film, bonded to a microfluidic channel, and interfaced to a time-of-flight mass spectrometer (TOFMS). A triangular-shaped thin polymer tip was formed by lithography and etching. A microfluidic channel, 20 microm wide and 10 microm deep, was embossed in a cyclo olefin substrate using a silicon master. The triangular tip was aligned with the channel and bonded between the channel plate and a flat plate to create a microfluidic channel with a wicking tip protruding from the end. This structure aided the formation of a stable Taylor cone at the apex of the tip, forming an electrospray ionization source. This source was tested by spraying several solutions for mass spectrometric analysis. Because the components are all made by lithographic approaches with high geometrical fidelity, an integrated array system with multiple channels can be formed with the same method and ease as a single channel. We tested a multichannel system in a multiplexed manner and showed reliable operation with no significant cross contamination between closely spaced channels.     

Negligible depletion solid-phase microextraction with radiolabeled analytes to study free concentrations and protein binding: an example with [3H]estradiol

A new method is presented that enables sensitive measurement of free concentrations of radiolabeled ligands. Additionally, protein binding of radiochemicals in complex matrixes can be determined with this new technique that combines negligible depletion solid-phase microextraction (nd-SPME) with liquid scintillation counting (LSC) as detection. [3H]Estradiol was taken as an example compound. Possible matrix effects of protein on fiber uptake kinetics were studied. No matrix effect was found, either by fouling of the fiber, or by changed uptake kinetics. The validity of the method was shown in the determination of the affinity constant (Ka) of estradiol for human serum albumin (HSA). The Ka was estimated at 8.9 x 10(4) M(-1), which corresponds well with literature values. This study shows that nd-SPME is suitable to study the free concentration and protein binding of [3H]estradiol. The method described in this paper combines the advantages of nd-SPME with the advantages of radiolabeled analytes, creating a timesaving, simple, and sensitive analytical tool that will be particularly useful in complex matrixes containing many potential interferences for chromatographic methods.     

Building blocks for a two-frequency laser lidar-radar: a preliminary study

A new principle of lidar-radar is theoretically and experimentally investigated. The proposed architecture is based on the use of an rf modulation of the emitted light beam and a direct detection of the backscattered intensity. Use of a radar-processing chain allows one to obtain range and Doppler measurements with the advantages of lidar spatial resolution. We calculate the maximum range of this device, taking into account different possible improvements. In particular, we show that use of a pulsed two-frequency laser and a spatially multimode optical preamplification of the backscattered light leads to calculated ranges larger than 20 km, including the possibility of both range and Doppler measurements. The building blocks of this lidar-radar are tested experimentally: The radar processing of an rf-modulated backscattered cw laser beam is demonstrated at 532 nm, illustrating the Doppler and identification capabilities of the system. In addition, signal-to-noise ratio improvement by optical pre-amplification is demonstrated at 1.06 microm. Finally, a two-frequency passively Q-switched Nd:YAG laser is developed. This laser then permits two-frequency pulses with tunable pulse duration (from 18 to 240 ns) and beat frequency (from 0 to 2.65 GHz) to be obtained.     

Off-axis measurements of atmospheric trace gases by use of an airborne ultraviolet-visible spectrometer

An airborne UV-visible spectrometer, the Gas Analyzer Spectrometer Correlating Optical Differences, airborne version (GASCOD/A4pi) was successfully operated during the Airborne Polar Experiment, Geophysica Aircraft in Antarctica airborne campaign from Ushuaia (54 degrees 49' S, 68 degrees 18' W), Argentina in southern spring 1999. The instrument measured scattered solar radiation through three optical windows with a narrow field of view (FOV), one from the zenith, two from the horizontal, as well as actinic fluxes through 2pi FOV radiometric heads. Only a few airborne measurements of scattered solar radiation at different angles from the zenith are available in the literature. With our configuration we attempted to obtain the average line-of-sight concentrations of detectable trace gases. The retrieval method, based on differential optical absorption spectroscopy, is described and results for ozone are shown and compared with measurements from an in situ instrument as the first method of validation.     

Amplitude and phase recovery of rotationally symmetric beams

Two methods are presented for the amplitude and phase recovery of optical beams with rotational symmetry. These are the tomographic method based on the ambiguity function and the one-step wavefront recovery based on the measurement of a phase-space distribution closely related to the Wigner distribution function. The results obtained from these two methods are compared, and the appropriateness of using either one of them for specific situations is discussed.     

Effect of suspended particulate-size distribution on the backscattering ratio in the remote sensing of seawater

Mie theory is used to study the influence of the particle-size distribution (PSD) on the backscattering ratio for case 1 and 2 waters. Several in situ measured PSDs from coastal water and the open ocean, representing typical case 2 and 1 waters, were used in this investigation. Calculation of the backscattering ratio requires integration of the PSD over a much broader size range than is usually measured. Consequently extrapolation from fitted data is necessary. To that purpose the measured data are fitted with hyperbolic (Junge) and the two-component model of the PSD. It is shown that the result of extrapolation, hence the backscattering ratio, critically depends on the chosen PSD model. For a particular PSD model the role of submicrometer particles and the applied integration limits on the backscattering ratio is discussed. The use of the hyperbolic PSD model largely overestimates the number of small (submicrometer) particles that significantly contribute to backscattering and consequently leads to an erroneously high backscattering ratio. The two-component model proves to be an adequate PSD model for use in backscattering/scattering calculations providing satisfactory results complying with experimental data. The results are relevant for the inversion of remotely sensed data and the prediction of optical properties and the concentration of phytoplankton pigments, suspended sediment, and yellow substance.     

Relaxed alignment tolerance of monolithic ring lasers

An investigation of relaxed alignment tolerance in monolithic ring lasers is presented. It was found that the relaxed alignment tolerance is strongly dependent on the out-of-plane angle. A preferred region for the out-of-plane angle is proposed. In this region the resonator has greatly relaxed alignment tolerances, and the unidirectional operation can be obtained under a much-lower magnetic field. Two lasers have been constructed in the region, and satisfactory performance was found, in good agreement with the theoretical analysis.     

Tunable green laser source based on frequency mixing of pump and laser radiation from a Nd:YVO4 crystal operating at 1342 nm with an intracavity KTP crystal

We report on cw tunable green laser light generation from a Nd:YVO4 laser operating at 1342 nm. Visible radiation was produced by a frequency mixing of pump and laser radiation inside an intracavity KTP crystal. When Nd:YVO4 was diode pumped, green laser was tunable from 503 to 505 nm. The green tuning range increases up to 22 nm when a Ti:sapphire laser is used as the pump source. In nonoptimal conditions, and for a pump power of 650 mW, green power was above 3 mW and close to 0.1 mW for Ti:sapphire and diode pumping, respectively.     

Resolution improvement in two-photon fluorescence microscopy with a single-mode fiber

The dependence of spectral broadening of an ultrashort-pulsed laser beam on the fiber length and the illumination power is experimentally characterized in order to deliver the laser for two-photon fluorescence microscopy. It is found that not only the spectral width but also the spectral blue shift increases with the fiber length and illumination power, owing to the nonlinear response in the fiber. For an illumination power of 400 mW in a 3-m-long single-mode fiber, the spectral blue shift is as large as 15 nm. Such a spectral blue shift enhances the contribution from the short-wavelength components within the pulsed beam and leads to an improvement in resolution under two-photon excitation, whereas the efficiency of two-photon excitation is slightly reduced because of the temporal broadening of the pulsed beam. The experimental measurement of the axial response to a two-photon fluorescence polymer block confirms this feature.