Fluorescent intensity

While a laser beam can be used for traditional absorption spectroscopy by measuring / and 7q, the strength of laser spectroscopy lies in more specialized experiments which often do not lend themselves to such measurements. Other techniques are connnonly used to detect the absorption of light from the laser beam. A coimnon one is to observe fluorescence excited by the laser. The total fluorescence produced is nonnally proportional to the amount of light absorbed. It can be used as a measurement of concentration to detect species present in extremely small amounts. Or a measurement of the fluorescence intensity as the laser frequency is scaimed can give an absorption spectrum. This may allow much higher resolution than is easily obtained with a traditional absorption spectrometer. In other experiments the fluorescence may be dispersed and its spectrum detennined with a traditional spectrometer. In suitable cases this could be the emission from a single electronic-vibrational-rotational level of a molecule and the experimenter can study how the spectrum varies with level. 

At low laser powers, the fluorescence signal is Imearly proportional to the power. Flowever, the power available from most tunable laser systems is suflFicient to cause partial saturation of the transition, with the result that the fluorescence intensity is no longer linearly proportional to the probe laser power. While more... 

Figure C 1.5.10. Nonnalized fluorescence intensity correlation function for a single terrylene molecule in p-terjDhenyl at 2 K. The solid line is tire tlieoretical curve. Regions of deviation from tire long-time value of unity due to photon antibunching (the finite lifetime of tire excited singlet state), Rabi oscillations (absorjDtion-stimulated emission cycles driven by tire laser field) and photon bunching (dark periods caused by intersystem crossing to tire triplet state) are indicated. Reproduced witli pennission from Plakhotnik et al [66], adapted from [118].

Single molecules also have promise as probes for local stmcture when doped into materials tliat are tliemselves nonfluorescent. Rlrodamine dyes in botli silicate and polymer tliin films exliibit a distribution of fluorescence maxima indicative of considerable heterogeneity in local environments, particularly for the silicate material [159]. A bimodal distribution of fluorescence intensities observed for single molecules of crystal violet in a PMMA film has been suggested to result from high and low viscosity local sites witliin tire polymer tliat give rise to slow and fast internal conversion, respectively [160]. 

Figure C2.3.18. Vibronic peak fluorescence intensity ratio (III/I) as a function of SDS concentration for 0.1 % PEO solutions o, —35 000 Daltons —600 000 Daltons). Open symbols are for aqueous solution without added salt, and filled symbols are for 100 mM aqueous NaCl. Reproduced with pennission from figure 2 of [111].

Nonradiative reiaxation and quenching processes wiii aiso affect the quantum yieid of fluorescence, ( )p = /cj /(/cj + Rsiative measurements of fluorescence quantum yieid at different quencher concentrations are easiiy made in steady state measurements absoiute measurements (to detemrine /cpjj ) are most easiiy obtained by comparisons of steady state fluorescence intensity with a fluorescence standard. The usefuiness of this situation for transient studies... 

The measurement of fluorescence intensity from a compound containing cliromophores of two spectral types is an example of a system for which it is reasonable to operate witli tire average rates of energy transfer between spectral pools of molecules. Let us consider tire simple case of two spectral pools of donor and acceptor molecules, as illustrated in figure C3.4.2 [18]. The average rate of energy transfer can be calculated as... 

Figure 9.43 The effect on time delay measurement of fluorescence intensity from sodium atoms in a transition state of Nal of changing the pump wavelength to (a) 300 nm, (b) 311 nm, (c) 321 nm, and (d) 339 nm. (Reproduced, with permission, from Rose, T. S., Rosker, M. J. and Zewail, A. H., J Chem. Phys., 91, 7415, 1989)...

Figure 9.45 (a) Oscillations, in the time domain, in the fluorescence intensity from the/state of I2... 

Electronic transitions in molecules in supersonic jets may be investigated by intersecting the jet with a tunable dye laser in the region of molecular flow and observing the total fluorescence intensity. As the laser is tuned across the absorption band system a fluorescence excitation spectrum results which strongly resembles the absorption spectrum. The spectrum... 

Unlike the situation regarding the crossing between the Vq and Fj potentials for Nal (see Figure 9.41), that for NaBr results in very efficient and rapid dissociation to give Na + Br when it is excited to Fj. Flow would you expect the fluorescence intensity from the neutral bromine atoms to vary with time compared with that for iodine atoms from Nal in Figure 9.42 ... 

Compai ison with literature experimental and calculation data showed that the model proposed ensures the accurate behavior of the functional dependence of x-ray fluorescence intensity on the particle size. Its main advantage is the possibility to estimate the effect of particle size for polydispersive multicomponent substances. 

Several methods were reported for the analysis of histamine, but the fluorimetric determination with o-phthaldialdehyde (OPA) the most widely used. It was shown that adducts, formed in the reaction of histamine with OPA in the presence of reducing agent, is more stable and gives high relative fluorescence intensity. The influences of different tiols on the fluorimeric determination histamine with OPA have been investigated. 

X-RAY FLUORESCENCE INTENSITY ELEMENTS OE MULTICOMPONENT POWDER MATERIAL WITH CONTINUOUS GRAIN SIZE DISTRIBUTION OE COMPONENTS... 

The existing models for emitting x-ray fluorescence intensity of elemental analytical lines from heterogeneous samples are limited in practical applications, because in most publications the relations between the fluorescence intensity of analytical lines elements and the properties of powder materials were not completely studied. For example, particles distribution of components within narrow layer of irradiator which emitted x-ray fluorescence intensity of elements might be in disagreement with particles distribution of components within whole sample. 

A pale yellow, crystalline product is obtained which fluoresces intense blue and gives the NMR results 42. Does the product have the desired structure ... 

Agglomerated impurities, such as particles or droplet residues, do not participate in the interference phenomenon leading to total reflection their fluorescence intensity is independent of the angle of incidence below the critical angle, and drops by a factor of 2 if the critical angle is surpassed due to the disappearance of the reflected component in the exciting beam nonreflecting impurities and residues). 

Experimental curves for the angular dependence of the fluorescence intensity from plated or sputtered submonatomic Ni layers (open triangles), layers produced by the evaporation of a Ni salt solution (open circles), and the silicon substrate (filled circles). 

Since then, TXRE has become the standard tool for surface and subsurface microanalysis [4.7-4.11]. In 1983 Becker reported the angular dependence of X-ray fluorescence intensities in the range of total reflection [4.12]. Recent demands have set the pace of further development in the field of TXRE - improved detection limits [4.13] in combination with subtle surface preparation techniques [4.14, 4.15], analyte concentrations extended even to ultratraces (pg) of light elements, e. g. A1 [4.16], spe-dation of different chemical states [4.17], and novel optical arrangements [4.18] and X-ray sources [4.19, 4.20]. 

The bulk type response curve depends also on surface roughness [4.34]. Reference materials must, therefore, be carefully investigated by angle-scan before use. Angle-scan characteristics of the sample, i.e. the fluorescence intensity recorded at more than one glancing angle near (j>, should not deviate from those of the reference. The measurement must be performed under similar optical conditions. 

Fig. 4.14. Fluorescence intensity from layers buried in a thick substrate. The dependence of intensity on the glancing angle was calculated for layers of different thickness but with a constant analyte area density. Silicon was assumed as substrate and Mo-Ka X-rays as primary beam. Total reflection occurs in the region below 0.1°. Without total reflection, the dashed horizontal line would be valid throughout [4.21].

Treatment of the chromatogram with a reagent results in the production of colored or fluorescent chromatogram zones, which are used to evaluate the success of the separation and for quantitative analysis For this purpose it is necessary that the color or fluorescence intensities remain stable for about 30 minutes... 

Fig. 51 Fluorescence intensities of porphyrins as a function of the concentration of the paraffin oil dipping solution mesoporphynn, coproporphynn, pentaporphynn...

Fig. 52 Fluorescence intensity of porphyrin chromatogram zones as a function of time after dipping in 50% liquid paraffin solution and storage in darkness. —...

Further examples pf fluorescence stabilization and intensity augmentation as a result of treatment of the chromatogram with viscous, lipophilic liquids are listed in Table 22. The alteration of the pH [293] or the addition of organic acids or bases [292] have also been found to be effective. Wintersteiger [291] has also described the effect that the TLC layer itself (binder) can influence the fluorescence intensity. 

Hypericin which is a hydroxyanthraquinone and the antibiotic nystatin also yield fluorescent zones. A higher fluorescence intensity is frequently obtained by heating to 88° C for 2 — 5 min instead of simply allowing to dry at room temperature. 

Immersion in a liquid paraffin - n-hexane (1 - - 3) did not lead to an appreciable increase in fluorescence intensity. 

The fluorescence intensity of the chromatogram zones could be stabilized and increased by a factor of 2.5 to 3.5 by subsequent immersion in liquid paraffin — -hexane (1 + 2). 

Note The developed chromatogram must be completely freed from nonpolar solvents before derivatization, otherwise an intense fluorescence will be stimulated over the whole plate. The fluorescence intensity of the chromatogram zones remains stable for ca. 40 min it decreases slowly as the layer dries out and can be returned to its original intensity by renewed immersion in the reagent solution or in water. 

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