Fluorescence, laser technique

In order to further extend the utility of fluorescence methods the use of time-resolution methods, fluorescence polarization, and laser techniques should be explored. The addition of other dyes with enhanced fluorescence properties on binding and increased selectivity to various types of nucleic acids will be necessary to further develop more useful analytical methods. 

In our previous paper (H), we introduced a novel experimental method to study the mechanistic details of solvent permeation into thin polymer films. This method incorporates a fluorescence quenching technique (19-20) and laser interferometry ( ). The former, in effect, monitors the movement of vanguard solvent molecules the latter monitors the dissolution process. We took the time differences between these two techniques to estimate both the nascent and the steady-state transition layer thicknesses of PMMA film undergoing dissolution in 1 1 MEK-isoproanol solution. The steady-state thickness was in good agreement with the estimate of Krasicky et al. (IS.). ... 

It is now possible to design the experiments using molecular beams and laser techniques such that the initial vibrational, rotational, translational or electronic states of the reagent are selected or final states of products are specified. In contrast to the measurement of overall rate constants in a bulk kinetics experiment, state-to-state differential and integral cross sections can be measured for different initial states of reactants and final states of products in these sophisticated experiments. Molecular beam studies have become more common, lasers have been used to excite the reagent molecules and it has become possible to detect the product molecules by laser-induced fluorescence . These experimental studies have put forward a dramatic change in experimental study of chemical reactions at the molecular level and has culminated in what is now called state-to-state chemistry. 

Laser microbeams offer several advantages over other fluorescence excitation techniques. In spectrofluorometry, observations are often made on a population of cells in a cuvette, resulting in a combined signal that lacks information about individual cellular responses. In flow cytometry, many individual cells are measured, but there is no temporal resolution since each cell is observed only once, and there is no spatial resolution since the entire cell is illuminated as it passes through the laser beam (see Chapter 30). In conventional fluorescence microscopy, individual cells can be monitored over time, and information about the two-dimensional spatial distribution of fluorescence can be obtained. However, some samples may be more susceptible to photobleaching by the arc lamps used for excitation, and the temporal resolution is limited to video-rate data acquisition (30 frames/s) (see Chapter 14). 

The IR spectra recorded by the fluorescence detection technique, where the population of a species in the ground state is monitored by laser-induced fluorescence with UV laser excitation, can be used successfully to detect dihydrogen bonds in the gas phase. It has been found, for example, that the v(O-H) and v(N-H) stretching vibrations in such IR experiments are very sensitive to dihydrogen bonding. 

Using this laser-induced fluorescence (LIF) technique, they have been able to determine branching ratios in the photolysis of various compounds. 

In conventional RE processes, the diffusive resistance is concentrated mainly inside the droplet, whereas the aqueous-side resistance can be neglected. This has been proven in Ref. 203 using the laser-induced-fluorescence (LIF) technique. Usually the organic phase is more viscous and the diffusion coefficients of the organic complexes are larger than those at the aqueous side, which supports this finding. 

In the gas-liquid two-phase flows illuminated by a laser sheet, for example, the intensity of light reflected from the gas-liquid interface (mostly the gas bubble s surface) not only saturate the CCD camera, but also overwhelm the intensity of light from the seeded tracer particles in its vicinity. Fluorescent particles are often used to realize the laser-induced fluorescence (LIF) technique together with PIV (e.g., Broder and Sommerfeld, 2002 Fujiwara et al., 2004a, b Kitagawa et al., 2005 Liu et al., 2005 Tokuhiro et al., 1998,1999), so that both images of gas-liquid interface (e.g., bubble s geometry) and velocity distribution in the liquid phase around the gas bubbles can be obtained. Issues on PIV measurement of gas-liquid two-phase flows will be further illustrated in the latter sections. 

The method proposed allows direct absolute measurement of local concentration at the instant of the laser pulse in a low pressure flame. We believe that this method could be applied to higher pressure flames by the use of ultrashort duration laser pulses with the new mode locked dye laser technique. But until the detector technology allows such short time resolutions we think that collisional lifetimes studies must be pursued to obtain more precise evaluation of the fluorescence efficiency, and to have a better understanding of the redistribution phenomena involved in optical pumping. For this purpose we are now studying the decay of resolved fluorescence lines. 

Bain, A.J. and McCaffery, A.J. (1985). On the measurement of molecular anisotropies using laser techniques. I. Polarized laser fluorescence, J. Chem. Phys., 83, 2627-2631. 

In the case of xanthone at least, this order is not only shown up in the Forster cycle estimates, but has been confirmed by observing the variation with pH of the optical densities of the triplet states of B and BH+ and comparing it with the fluorescence intensity behaviour (see Fig. 6). Confirmation that the pK order obtained using the Forster cycle is reliable in such cases is also found in a direct determination of p/ (Tj) of benzophenone by a laser technique the value derived is consistent with earlier phosphorescence observations (Rayner and Wyatt, 1974). Ledger and Porter (1972) observed a marked decrease in the phosphorescence intensity of benzophenone near pH 5, and the apparent discrepancy between this result and the p/ (Tj )-value of 1-5 is due to the very large difference in lifetimes of BH+(T,) and B(Tj). Since unprotonated benzophenone has a very shortlived St state [1/kj for the intersystem crossing alone in ethanol is 16 5 ps (Hochstrasser et al., 1974)], protonation in this state is unlikely. However, Forster cycle calculations indicate that the singlet state would be a weaker base than the triplet state. The realization that unprotonated benzaldehyde and acetophenone had Tj states of the... 

The jet-cooled species can be studied using standard laser techniques (Figure 13-4a) like laser-induced fluorescence (LIF or fluorescence excitation spectroscopy) or R2PI, as well as using their sophisticated double-resonance variants, like UV/UV or IR/UV double resonance spectroscopy (Figure 13-4b and 4c), to investigate molecules or clusters existing under various isomeric forms... 

Radicals.—The measurement of emission intensities from electronically excited small free radicals has become an important means of determining radical concentrations in hostile environments such as flames. When combined with laser excitation, the technique is very powerful, offering temporal, spectral, and spatial resolution. Just has reviewed laser techniques for the measurement of both radical concentrations and local temperatures in flames, and has demonstrated the use of laser-induced saturated fluorescence to measure the concentrations of CH and OH radicals in low-pressure acetylene-oxygen flames. Vanderhoff ei al. used a novel Kr " and Ar laser intracavity technique to... 

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