Excitation energy transfer quenching method

The structural dimension at a water/DCE interface is d — 2.48, while short-range structural information about the interface obtained by the fluorescence dynamic anisotropy experiments suggests that the interface is three-dimensional-like. Taking the results obtained by molecular dynamics simulations into account, these results can be understood only by the fact that the water/DCE interface is thin ( 1 nm), but is rough with respect to the spatial resolution of the excitation energy transfer quenching method ( 7 nm), as shown in Figure 12.7. 

The most important applications of luminescence probing in microemulsions involve the deactivation dynamics or excitation energy transfer properties of the excited states. With a brief flash of light a population of excited species is created in the sample, and the subsequent deactivation is observed over time. The decay of the excited probe, and the fluorescence spectrum, may depend on the interactions with the environment, which reveal useful information. In time-resolved luminescence quenching (TRLQ), however, it is the interaction of the probe with another added component, a quencher, that is studied. This method is dealt with here. For micellar systems, several publications have already discussed it in both experimental and theoretical detail [1-6]. 

Donor quenching is the most common method used for detecting energy transfer. Quenching of the donor fluorescence is due to acceptors at different distances as well as orientations, and due to the motions of donor or acceptor. Excitation is set at the wavelength of donor absorption and the emission of the donor is monitored. The emission wavelength of donor is selected such that uo coutrihution from the acceptor fluorescence is observed. 

Additionally, since the acceptor is excited as a result of FRET, those acceptors that are fluorescent will emit photons (proportional to their quantum efficiency) also when FRET occurs. This is called sensitized emission and can also be a good measure of FRET (see Fig. 1). To quantitate FRET efficiency in practice, several approaches have been evolved so far. In flow cytometric FRET (7), we can obtain cell-averaged statistics for large cell populations, while the subcellular details can be investigated with various microscopic approaches. Jares-Erijman and Jovin have classified 22 different approaches that can be used to quantify energy transfer (8). Most of them are based on donor quenching and/or acceptor sensitization, and a few on measuring emission anisotropy of either the donor or the acceptor. Some of these methods can be combined to extend the information content of the measurement, for example two-sided FRET (9) involves both acceptor depletion (10) and... 

COLLISIONAL ENERGY-TRANSFER SPECTROSCOPY WITH LASER-EXCITED ATOMS IN CROSSED ATOM BEAMS A NEW METHOD FOR INVESTIGATING THE QUENCHING OF ELECTRONICALLY EXCITED ATOMS BY MOLECULES... 

Collisional Energy-transfer Spectroscopy with Laser-excited Atoms in Crossed Atom Beams A New Method for Investigating the Quenching of Electronically Excited Atoms by Molecules... 

With site-directed mutation and femtosecond-resolved fluorescence methods, we have used tryptophan as an excellent local molecular reporter for studies of a series of ultrafast protein dynamics, which include intraprotein electron transfer [64-68] and energy transfer [61, 69], as well as protein hydration dynamics [70-74]. As an optical probe, all these ultrafast measurements require no potential quenching of excited-state tryptophan by neighboring protein residues or peptide bonds on the picosecond time scale. However, it is known that tryptophan fluorescence is readily quenched by various amino acid residues [75] and peptide bonds [76-78]. Intraprotein electron transfer from excited indole moiety to nearby electrophilic residue(s) was proposed to be the quenching... 

The efficiency of quenching is proportional to the rate constant for energy transfer, kq, and the lifetime r of the excited triplet being quenched. Since kq values are so close to being diffusion controlled, quenching studies provide a simple method for measuring triplet lifetimes. 

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