Femtosecond fluorescence upconversion measurements

Sample preparation was given elsewhere [2]. Femtosecond fluorescence upconversion and picosecond time-correlated single-photon-counting set-ups were employed for the measurement of the fluorescence transients. The system response (FWHM) of the femtosecond fluorescence up-conversion and time-correlated single-photon-counting setups are 280 fs and 16 ps, respectively [3] The measured transients were fitted to multiexponential functions convoluted with the system response function. After deconvolution the time resolution was 100 fs. In the upconversion experiments, excitation was at 350 nm, the transients were measured from 420 nm upto 680 nm. Experiments were performed under magic angle conditions (to remove the fluorescence intensity effects of rotational motions of the probed molecules), as well as under polarization conditions in order to obtain the time evolution of the fluorescence anisotropy. 

To reveal possible ultrafast processes occurring on a time scale less than 30 ps, femtosecond fluorescence upconversion experiments were performed [30] in toluene under magic angle polarization. To extract complete information of the decay times and their amplitudes in function of detection wavelength, the measurements were performed in three time windows of 5 ps, 50 ps, and 420 ps. 

The short time scale dynamics have been studied by means of femtosecond fluorescence upconversion. For all dendrimers these measurements revealed size-independent kinetic processes related to an internal vibrational redistribution, a vibrational/solvent relaxation. Singlet-singlet annihilation, only present in the multichromophoric compounds, was established by an excitation energy-dependent study. It has been shown that this type of process contributes to a larger extent in the para-substituted dendrimers compared to the meta-substituted ones. These differences between the meta- and para-substituted dendrimers... 

Precise measurements of the excited state lifetimes of the DNA constituents were not available till very recently, mainly due to the limited time resolution of conventional spectroscopic techniques. Studying the DNA nucleosides by transient absorption spectroscopy, Kohler and co-workers observed a very short-lived induced absorption in the visible which they assigned to the first excited state [5,6]. The lifetimes observed were all well below 1 picosecond. The first femtosecond fluorescence studies of DNA constituents were performed using the fluorescence upconversion technique. Peon and Zewail [7] reported that the excited state lifetimes of DNA/RNA nucleosides and nucleotides all fall in the subpicosecond time, thus corroborating the results obtained by transient absorption. 

Solvation dynamics are measured using the more reliable energy relaxation method after a local perturbation [83-85], typically using a femtosecond-resolved fluorescence technique. Experimentally, the wavelength-resolved transients are obtained using the fluorescence upconversion method [85], The observed fluorescence dynamics, decay at the blue side and rise at the red side (Fig. 3a), reflecting typical solvation processes. The molecular mechanism is schematically shown in Fig. 5. Typically, by following the standard procedures [35], we can construct the femtosecond-resolved emission spectra (FRES, Stokes shifts with time) and then the correlation function (solvent response curve) ... 

The methods discussed so far, fluorescence upconversion, the various pump-probe spectroscopies, and the polarized variations for the measurement of anisotropy, are essentially conventional spectroscopies adapted to the femtosecond regime. At the simplest level of interpretation, the information content of these conventional time-resolved methods pertains to populations in resonantly prepared or probed states. As applied to chemical kinetics, for most slow reactions (on the ten picosecond and longer time scales), populations adequately specify the position of the reaction coordinate intermediates and products show up as time-delayed spectral entities, and assignment of the transient spectra to chemical structures follows, in most cases, the same principles used in spectroscopic experiments performed with continuous wave or nanosecond pulsed lasers. 

With the development of fluorescence upconversion techniques, which nowadays provide femtosecond time resolution, it is also possible to directly measure the time evolution of the spontaneous emission following the excitation of the sample by the pirnip pulse. In this method, the fluorescence is collected and focused onto a nonlinear crystal, where it is superposed with the probe beam in order to perform upconversion. Time resolution is achieved because the probe pulse creates a time gate for the spontaneous emission, i.e. the fluorescence is only measured within the duration of the probe. Frequency resolution is achieved by subsequently dispersing the upconverted signal in a monochromator. Although fluorescence detection provides less photon yield than stimulated techniques, it has the desirable feature to exclusively monitor the time evolution in the initially excited electronic states (cf. the discussion above). 

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