Fluorescence life time

Various spectroscopic techniques and probes have been used to investigate solubilization of probe molecules, mostly using UV/visible spectroscopy, fluorescence spectroscopy, ESR spectroscopy [64, 74, 217, 287] and NMR-spectro-scopy [367-369]. Fluorescence spectroscopy is particularly versatile [370], as various static and dynamic aspects can be covered by studying excitation and emission spectra, excimer or exciplex formation, quantum yields, quenching, fluorescence life-times, fluorescence depolarization, energy transfer etc. 

The ESR spectrum of the pyridazine radical anion, generated by the action of sodium or potassium, has been reported, and oxidation of 6-hydroxypyridazin-3(2//)-one with cerium(IV) sulfate in sulfuric acid results in an intense ESR spectrum (79TL2821). The self-diffusion coefficient and activation energy, the half-wave potential (-2.16 eV) magnetic susceptibility and room temperature fluorescence in-solution (Amax = 23 800cm life time 2.6 X 10 s) are reported. 

Because luciferyl adenylate emitted a red chemiluminescence in the presence of base, coinciding with the red fluorescence of 5,5-dimethyloxylucferin, the keto-form monoanion Cl in its excited state is considered to be the emitter of the red light. Thus, the emitter of the yellow-green light is probably the enol-form dianion C2 in its excited state, provided that the enolization takes place within the life-time of the excited state. Although the evidence had not been conclusive, especially on the chemical structures of the light emitters that emit two different colors, the mechanism shown in Fig. 1.12 was widely believed and cited until about 1990. 

The measurement of fluorescence life-times has great value for probing structural features of proteins. It requires expensive equipment since very rapid extinction of the exciting nanosecond pulse is necessary and the rapid decay of the emission must then be measured. The decay of the tryptophan fluorescence of LADH is biphasic with x = 3.9 and 7.2 ns and these are assigned to buried Trp-314 and exposed Trp-15, respectively. ... 

Proteases have received less attention than lipases, but in one of the earliest papers on biocatalysis in ionic liquids it was noted that the activity loss of thermo-lysin during preincubation proceeded much more slowly in [BMIm][PF6] than in ethyl acetate [8]. The storage stability of a-chymotrypsin in the ionic liquid [EMIm][ Tf2N] was compared with that in water, 3 M sorbitol, and 1-propanol. The residual hydrolytic activity (after dilution with aqueous buffer) was measured vs time, and structural changes were monitored by fluorescence and CD spectroscopy as well as DSC [98]. The enzyme s life-time in [EMIm][ Tf2N] at 30°C was more than twice that in 3 M sorbitol, six times as long as that in water, and 96 times as long as that in 1-propanol. 

Phase sensitivity A phase-sensitive flow cytometer quantifies the life time of the fluorescence emitted by particles. The decay time of fluorescence from a given fluorochrome is altered by changing chemical environments, and therefore measurement of fluorescence lifetimes can provide information about the microenvironment surrounding the fluorescent probe. 

The fluorescence probe l-anilinonaphthalene-8-sulfonate was used to examine the binding of spin-labeled local anesthetics to membranes of human blood cells and rabbit sarcoplasmic reticulum. Two distinct fluorescence lifetimes are exhibited by this probe the shorter life time represents the probe associated with pure lipid, the... 

The physical basis of the second type of approach rests upon the effect of the local electrostatic potential upon dynamic interactions at encounters with charged quenching molecules resulting in fluorescence (phosphorescence) (Vogel et al., 1986 Anni et al., 1994) or between a stable radical, e.g. nitroxide, and another charged paramagnetic species (Likhtenshtein et al., 1972 Likhtenshtein, 1976, 1988,1993). In such cases, the relaxation parameters, i.e. the life-time of the fluorescence (phosphorescence) chromophore or spin-spin and spin-lattice relaxation rates of paramagnetic species are dependent upon the frequency of encounters, and, therefore, on local electrostatic fields... 

Fluorescence-based detection methods are the most commonly used readouts for HTS as these readouts are sensitive, usually homogeneous and can be readily miniaturised, even down to the single molecule level.7,8 Fluorescent signals can be detected by methods such as fluorescence intensity (FI), fluorescence polarisation (FP) or anisotropy (FA), fluorescence resonance energy transfer (FRET), time-resolved fluorescence resonance energy transfer (TR-FRET) and fluorescence intensity life time (FLIM). Confocal single molecule techniques such as fluorescence correlation spectroscopy (FCS) and one- or two-dimensional fluorescence intensity distribution analysis (ID FID A, 2D FIDA) have been reported but are not commonly used. 

Examinations of other physical properties of pyridazine include the ESR spectrum of pyridazine radical anion (obtained with pyridazine and sodium or potassium in dimethoxyethane or tetra-hydrofuran, the self-diffusion coefficient and activation energy, the half-wave potential (-2.16V), and magnetic susceptibility. Pyridazine was reported not to fluoresce and no luminiscence could be observed even under very long exposures. More recently, room-temperature fluorescence in solution is reported to be at 23,800 cm (max.), with a life time of 2.6 x 10 . ... 

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