Fluorescence depletion spectroscopy

Femtosecond Time-Resolved Fluorescence Depletion Spectroscopy 321... 

Kauffman, J.F., Cote, M.J., Smith, P.G., McDonald, J.D. Picosecond fluorescence depletion spectroscopy. 2. Intramolecular vibrational relaxation in the excited electronic state of fluorene. J. Chem. Phys. 90, 2874-2891 (1989)... 

The structure of the 1 1 BA-H2O complex in its ground state has been determined by rotational coherence spectroscopy (RCS) using the fluorescence depletion scheme [74, 75], The empirical minimum-energy calculation supplemented the insufficient information obtained from the RCS measurement [74], In the structure giving the best fit to the RCS signal, the water molecule sits on the terminal aromatic ring of the one anthracene moiety. Thus the two anthracene moieties are stabilized by the water molecule in a different manner, and such asymmetry in the stabilization, i.e., symmetry breaking, facilitates the electron jump from one anthracene to the other [76]. 

The nanosized detection area Ar or volume created by STED also extends the power of fluorescence correlation spectroscopy (FCS) and the detection of molecular diffusion [74,95]. For example, STED microscopy has probed the diffusion and interaction of single lipid molecules on the nanoscale in the membrane of a living cell (Fig. 19.6). The up to 70 times smaller detection areas created by STED (as compared to confocal microscopy) revealed marked differences between the diffusion of sphingo- and phospholipids [74]. While phospholipids exhibited a comparatively free diffusion, sphingolipids showed a transient ( 10 ms) cholesterol-mediated trapping taking place in a < 20-nm diameter area, which disappeared after cholesterol depletion. Hence, in an unperturbed cell putative cholesterol-mediated lipid membrane rafts should be similarly short-lived and smaller. 

Yang, X., Dagdigian, P.J. Fluorescence excitation and depletion spectroscopy of the BAr complex Electronic states correlating with the excited valence B(2s2p D) asymptote, J. Chem. Phys. 106 (1997) 6596-6606. 

Fourier transform infrared (FTIR) spectroscopy, 13C nuclear magnetic resonance (NMR) spectroscopy, ultraviolet-visible (UV-VIS) and fluorescence spectroscopy can be integrated with chromatographic techniques especially in the study of ageing and degradation of terpenic materials. They can be used to study the transformation, depletion or formation of specific functional groups in the course of ageing. 

Much of the early studies of surfactant adsorption at the solid-solution interface were based on classical experimental techniques, such as solution depletion [1, 32], fluorescence spectroscopy [2], and measurements of the differential enthalpy of adsorption [2], Such methods have provided much of the basic initial understanding. However, they provide no direct structural information and are difficult to apply to mixtures [23, 34], However, when combined with other techniques, such as NMR and flow microcalorimetry, they provide some insight into the behaviour of mixtures. This was demonstrated by Thibaut et al. [33] on SDS/C10E5 mixtures adsorbed onto silica and by Colombie et al. [34] on the adsorption of SLS/Triton X-405 mixtures onto polystyrene particles. 

Figure 12-1. Schematic diagram to illustrate double resonance techniques, (a) REMPI 2 photon ionization. The REMPI wavelength is scanned, while a specific ion mass is monitored to obtain a mass dependent SI <- SO excitation spectrum, (b) UV-UV double resonance. One UV laser is scanned and serves as a burn laser, while a second REMPI pulse is fired with a delay of about 100 ns and serves as a probe . The probe wavelength is fixed at the resonance of specific isomer. When the burn laser is tuned to a resonance of the same isomer it depletes the ground state which is recorded as a decrease (or ion dip) in the ion signal from the probe laser, (c) IR-UV double resonance spectroscopy, in which the burn laser is an IR laser. The ion-dip spectrum reflects the ground state IR transitions of the specific isomer that is probed by the REMPI laser, (d) Double resonance spectroscopy can also use laser induced fluorescence as the probe, however that arrangement lacks the mass selection afforded by the REMPI probe...

All these studies with femtosecond pulses on the primary photochemical processes of rhodopsin were done by means of transient absorption (pump probe) spectroscopy [10]. However, absorption spectroscopy may not be the best way to probe the excited-state dynamics of rhodopsin, because other spectral features, such as ground-state depletion and product absorption, are possibly superimposed on the excited-state spectral features (absorption and stimulated emission) in the obtained data. Each spectral feature may even vary in the femtosecond time domain, which provides further difficulty in analyzing the data. In contrast, fluorescence spectroscopy focuses only on the excited-state processes, so that the excited-state dynamics can be observed more directly. 

We would like to mention one further practical application of standard Raman spectroscopy, namely the method of Raman lidar, which is now routinely used to monitor the upper atmosphere for composition (e.g. the presence of water vapour), chemical processes (e.g. the generation or depletion of ozone (O3)), and the determination of temperature profiles at high altitudes. Although absorption and fluorescence lidar systems are also widely used, Raman lidar has the distinct advantage that it is a simultaneous multispecies measurement technique, and that only a single fixed-wavelength laser is required. 

Structure. As for ground state IR-UV ion dip spectroscopy, the depletion in the electronically excited state caused by IR absorption can also be probed via LIF. LIF detection should be equally applicable to ground state and excited states, but has more limited application as strongly fluorescent molecules are required [102, 104, 105]. 

This optical-optical double-resonance technique has already been used for other Doppler-free techniques [10.25], such as polarization spectroscopy (see Sect.10.3). Its applications to molecular beams has, however, the following advantages compared to spectroscopy in gas cells. When the chopped pump laser periodically depletes the level E. and populates level Ej, there are two relaxation mechanisms in gas cells which may transfer the population modulation to other levels. These are collision processes and laser-induced fluorescence (see Fig.8.39). The neighboring levels therefore also show a modulation and the modulated excitation spectrum induced by the probe laser includes all lines which are excited from those levels. If several absorption lines overlap within their Doppler width, the pump laser simultaneously excites several upper states and also partly depletes several lower levels. 

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