False estimates of stimulated Raman pumping efficiency caused by the optical Stark effect.

One technique for measuring the fraction of molecules pumped to the excited state in stimulated Raman pumping (SRP) is to record the depletion of molecules in the lower state by resonance enhanced multiphoton ionization (REMPI). The presence of electric fields on the order of 10(7) V/cm arising from the pulsed SRP laser beams is sufficient to shift the line position of the REMPI transition to such an extent that the estimate of the pumping efficiency is overestimated unless this shift is accounted for.

Laser chemical analysis.

Selected applications of laser methods to analytical problems are reviewed. Examples are chosen from multiphoton ionization and laser fluorescence analysis. Although efforts to carry out elemental analysis with laser techniques are probably the most advanced, studies suggest that the analysis of molecular species is also quite promising, particularly in regard to interfacing laser fluorimetric detection with high-performance liquid chromatography. Recent experiments indicate that analysts can expect to attain in a number of cases the ultimate limit of single-atom or single-molecule detection with laser-based methods.

Probing individual molecules with confocal fluorescence microscopy.

Confocal fluorescence microscopy coupled with a diffraction-limited laser beam and a high-efficiency detection system has been used to study the diffusive movement and emission process of individual fluorescent molecules in the liquid phase at room temperature. The high detection sensitivity achieved at fast data acquisition speeds (greater than 1 kilohertz) allows real-time observation of single-molecule fluorescence without statistical analysis. The results show fluorescence-cycle saturation at the single-molecule level and multiple recrossings of a single molecule into and out of the probe volume as well as the triplet state.

Laser fluorimetry: subpicogram detection of aflatoxins using high-pressure liquid chromatography.

The use of high-pressure liquid chromatographic separation in conjunction with laser-induced fluorescence detection permits the analysis of trace fluorescent species at new limits of sensitivity. This technique was applied to the carcinogens aflatoxins B1, B2, G1, and G2, which were linearly quantitated to 7.5 x 10(-13) gram. The procedure consists of forming more fluorescent aflatoxin derivatives, eluting the aflatoxins from a reverse-phase column, focusing the 325-nanometer output of a helium-cadmium ion laser into a suspended droplet of the eluent, and measuring the resulting fluorescence using phase-sensitive detection.

Electrokinetic separation of chiral compounds.

Femtomole amounts of racemic mixtures of derivatized amino acids were resolved and analyzed rapidly in about 10 minutes by means of high-voltage zone electrophoresis with laser-fluorescence detection. The electrophoresis was performed in capillary columns containing a chiral support electrolyte. A number of dansyl amino acids were resolved by the diastereomeric interaction between the DL-amino acid and the copper(II) complex of L-histidine present in the support electrolyte. A combination of electro-osmotic and electrophoretic action caused all species, positively charged, neutral, and negatively charged, to pass through the 0.5- nanoliter detection volume where they were subjected to laser excitation.

Application of two-step laser mass spectrometry to cosmogeochemistry: direct analysis of meteorites.

Polycyclic aromatic hydrocarbons in C1, C2, and C3 carbonaceous chondrites and in some ordinary chondrites have been directly analyzed by two-step laser desorption/ laser multiphoton ionization mass spectrometry, a selective and sensitive method requiring only milligram samples. At the ionization wavelength of 266 nanometers, parent ion peaks of polycyclic aromatic hydrocarbons dominate the mass spectra. Quantitative analysis is possible; as an example, the concentration of phenanthrene in the Murchison meteorite was determined to be 5.0 parts per million.

Capillary electrophoresis.

Electrophoretic separations carried out in capillary tubes offer the possibilities of rapid and automated analyses of small volumes of complex mixtures with unprecedented resolution and sensitivity. Some emerging developments of this new instrumentation are reviewed.

Partial control of an ion-molecule reaction by selection of the internal motion of the polyatomic reagent ion.

The ion-molecule reaction NH(3)(+) + ND(3) has been studied at various collision energies (1 to 5 electron volts in the center of mass) with preparation of the NH(3)(+) reagent in two nearly isoenergetic vibrational states. One state corresponds to pure out-of-plane bending of the planar NH(3)(+) ion (0.60 electron volts), whereas the other state is a combination of in-plane and out-of-plane motion (0.63 electron volts). The product branching ratios differ markedly for these two vibrational-state preparations. The differences in reactivity suggest that the in-plane totally symmetric stretching mode is essentially inactive in controlling the branching ratio of this reaction.

Spatially resolved organic analysis of the allende meteorite.

The distribution of polycyclic aromatic hydrocarbons(PAHs) in the Allende meteorite has been probed with two-step laser desorption/laser multiphoton ionization mass spectrometry. This method allows direct in situ analysis with a spatial resolution of 1 square millimeter or better of selected organic molecules. Spectra from freshly fractured interior surfaces of the meteorite show that PAH concentrations are locally high compared to the average concentrations found by wet chemical analysis of pulverized samples. The data suggest that the PAHs are primarily associated with the fine-grained matrix, where the organic polymer occurs. In addition, highly substituted PAH skeletons were observed. Interiors of individual chondrules were devoid of PAHs at our detection limit(about 0.05 parts per million).

Single cells as biosensors for chemical separations.

A biosensor system based on the response of living cells was demonstrated that can detect specific components of a complex mixture fractionated by a microcolumn separation technique. This system uses ligand-receptor binding and signal-transduction pathways to biochemically amplify the presence of an analyte after electrophoretic separation. The transduced signal was measured by means of two approaches: (i) fluorescence determination of intracellular calcium concentrations in one or more rat PC-12 cells and (ii) measurement of transmembrane current in a Xenopus laevis oocyte microinjected with messenger RNA that encodes a specific receptor. This analysis system has the potential to identify biologically active ligands present in a complex mixture with exceptional sensitivity and selectivity.

Reaction product imaging: the h + d2 reaction.

The differential cross section for the H + D(2) --> HD + D reaction has been measured using a technique called reaction product imaging. In this experiment, a photolytically produced beam of hydrogen (H) atoms crossed a beam of cold deuterium (D(2)) molecules. Product D atoms were ionized at the intersection of the two particle beams and accelerated toward a position-sensitive detector. The ion images appearing on the detector are two-dimensional projections of the three-dimensional velocity distribution of the D atom products. The reaction was studied at nominal center-of-mass collision energies of 0.54 and 1.29 electron volts. At the lower collision energy, the measured differential cross section for D atom production, summed over all final states of the HD(v,J) product, is in good agreement with recent quasi-classical trajectory calculations. At the higher collision energy, the agreement between the theoretical predictions and experimental results is less favorable.

Identification of complex aromatic molecules in individual interplanetary dust particles.

Seventeen stratospherically collected particles-eight of which are classified as interplanetary dust particles (IDPs), seven of which are classified as probable terrestrial contaminants, and two of which have uncertain origins-were studied with a microprobe two-step laser mass spectrometer. Many polycyclic aromatic hydrocarbons(PAHs) and their alkylated derivatives were identified in two of the eight IDPs. The PAHs observed include a high-mass envelope not found in meteorites or terrestrial contaminants and prominent odd-mass peaks suggestive of nitrogen-containing functional groups attached to aromatic chromophores. In addition, the complexity of the IDP mass spectra has no precedence in previous studies of meteorite samples or their acid residues. Extensive checks were performed to demonstrate that the PAH signals are not caused by terrestrial contaminants.

Boundary layer profiles in plasma chemical vapor deposition.

A nonlinear optical spectroscopy based on degenerate four-wave mixing has made possible direct measurements of species temperature and concentration profiles through the boundary layer of a reactive plasma at atmospheric pressure. Spectra were obtained for CH and C(2) radicals over a range of conditions including those for the plasma chemical vapor deposition of diamond films. Numerical simulations based on a one-dimensional stagnation-point flow model are in good agreement with the measurements. The CH mole fraction is shown to rise and fall as a function of distance from the substrate, which is compelling experimental evidence for the complex chemistry that is occurring in the plasma boundary layer.

State-to-State Rates for the D + H2(v = 1, j = 1) rarr HD(v', j') + H Reaction: Predictions and Measurements.

A fully quantal wavepacket approach to reactive scattering in which the best available H(3) potential energy surface was used enabled a comparison with experimentally determined rates for the D + H(2)(v = 1, j = 1) --> HD(v' = 0, 1, 2; j') + H reaction at significantly higher total energies (1.4 to 2.25 electron volts) than previously possible. The theoretical results are obtained over a sufficient range of conditions that a detailed simulation of the experiment was possible, thus making this a definitive comparison of experiment and theory. Good to excellent agreement is found for the vibrational branching ratios and for the rotational distributions within each product vibrational level. However, the calculated rotational distributions are slightly hotter than the experimentally measured ones. This small discrepancy is more marked for products for which a larger fraction of the total energy appears in translation. The most likely explanation for this behavior is that refinements are needed in the potential energy surface.

UV irradiation of polycyclic aromatic hydrocarbons in ices: production of alcohols, quinones, and ethers.

Polycyclic aromatic hydrocarbons (PAHs) in water ice were exposed to ultraviolet (UV) radiation under astrophysical conditions, and the products were analyzed by infrared spectroscopy and mass spectrometry. Peripheral carbon atoms were oxidized, producing aromatic alcohols, ketones, and ethers, and reduced, producing partially hydrogenated aromatic hydrocarbons, molecules that account for the interstellar 3.4-micrometer emission feature. These classes of compounds are all present in carbonaceous meteorites. Hydrogen and deuterium atoms exchange readily between the PAHs and the ice, which may explain the deuterium enrichments found in certain meteoritic molecules. This work has important implications for extraterrestrial organics in biogenesis.

Patch-clamp detection of neurotransmitters in capillary electrophoresis.

Gamma-aminobutyrate acid, L-glutamate, and N-methyl-D-aspartate were separated by capillary electrophoresis and detected by the use of whole-cell and outside-out patch-clamp techniques on freshly dissociated rat olfactory interneurons. These neuroactive compounds could be identified from their electrophoretic migration times, unitary channel conductances, and power spectra that yielded corner frequencies and mean single-channel conductances characteristic for each of the different agonist-receptor interactions. This technique has the sensitivity to observe the opening of a single ion channel for agonists separated by capillary electrophoresis.

Probing single secretory vesicles with capillary electrophoresis.

Secretory vesicles obtained from the atrial gland of the gastropod mollusk Aplysia californica were chemically analyzed individually with a combination of optical trapping, capillary electrophoresis separation, and a laser-induced fluorescence detection. With the use of optical trapping, a single vesicle that had attoliters (10(-18) liters) of volume was introduced into the tapered inlet of a separation capillary. Once the vesicle was injected, it was lysed, and its components were fluorescently labeled with naphthalene-2, 3-dicarboxaldehyde before separation. The resultant electropherograms indicated distinct variations in the contents of single vesicles.

Chemical transformations in individual ultrasmall biomimetic containers.

Individual phospholipid vesicles, 1 to 5 micrometers in diameter, containing a single reagent or a complete reaction system, were immobilized with an infrared laser optical trap or by adhesion to modified borosilicate glass surfaces. Chemical transformations were initiated either by electroporation or by electrofusion, in each case through application of a short (10-microsecond), intense (20 to 50 kilovolts per centimeter) electric pulse delivered across ultramicroelectrodes. Product formation was monitored by far-field laser fluorescence microscopy. The ultrasmall characteristic of this reaction volume led to rapid diffusional mixing that permits the study of fast chemical kinetics. This technique is also well suited for the study of reaction dynamics of biological molecules within lipid-enclosed nanoenvironments that mimic cell membranes.

Search for past life on Mars: possible relic biogenic activity in martian meteorite ALH84001.

Fresh fracture surfaces of the martian meteorite ALH84001 contain abundant polycyclic aromatic hydrocarbons (PAHs). These fresh fracture surfaces also display carbonate globules. Contamination studies suggest that the PAHs are indigenous to the meteorite. High-resolution scanning and transmission electron microscopy study of surface textures and internal structures of selected carbonate globules show that the globules contain fine-grained, secondary phases of single-domain magnetite and Fe-sulfides. The carbonate globules are similar in texture and size to some terrestrial bacterially induced carbonate precipitates. Although inorganic formation is possible, formation of the globules by biogenic processes could explain many of the observed features, including the PAHs. The PAHs, the carbonate globules, and their associated secondary mineral phases and textures could thus be fossil remains of a past martian biota.

Correction: Effects of reagent rotation on interferences in the product angular distributions of chemical reactions.

[This corrects the article DOI: 10.1039/C5SC03373J.].