Lasers with absorption fluorescence

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. 

In order to prepare successful NIR molecular probe dyes, NIR dyes must meet the following criteria adequate response to analytes, high lipophilicity and/or reactive functional groups, absorbance maxima compatible with available laser diodes, high fluorescence quantum yield, molar absorptivity, and high photostability. 

When the frequency of a laser falls fully into an absorption band, multiple phonon processes start to appear. Leite et al 2° ) observed /7 h order ( = 1, 2. 9) Raman scattering in CdS under conditions of resonance between the laser frequency and the band gap or the associated exciton states. The scattered light spectrum shows a mixture of fluorescent emission and Raman scattering. Klein and Porto 207) associated the multiphonon resonance Raman effect with the fluorescent emission spectrum, and suggested a possible theoretical approach to this effect. 

A novel pump-damp-probe method (PDPM), which allows the characterization of solvation dynamics of a fluorescence probe not only in excited but also in the ground states has been recently developed (Changenet-Barret, 2000 and references therein). In PDPM, a pump produces a nonequilibrium population of the probe excited, which, after media relaxation, is simulated back to the ground states. The solvent relaxation of the nonequlibrium ground state is probed by monitoring with absorption technique. The inramolecular protein dynamics in a solvent-inaccessible region of calmodulin labeled with coumarin 343 peptide was examined by PDPM. In the pump-dump-probe experiments, part of a series of laser output pulses was frequency-doubled and softer beams were used as the probe. The delay of the probe with respect to the pump was fixed at 500 ps. 

Due to its relevance to the next section, we observed and analyzed the fluorescent emission of Tryptophan in water solution excited by one, two, and three-photon absorption. For that, three different light sources were used a UV (180-375 nm) lamp, the second harmonic of a Q-switched Nd YAG laser (with 8 ns pulse duration at 532 nm) and a Ti-Sapphire laser delivering pulses at 76 MHz, with 150 fs pulse duration and 500 mW average power at 800 nm. 

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