Resolved Emission Spectra

Suppose the total inleosity decr s with a single decay time T. The center of gravity observed in asteady-state emisrion spectrum is given by the integral average overthe 

This expression connects the center of gravity observed in a steady-state experiment with the relative values of the decay time t and the spectral relaxation time Xs. From this expression one can also understand the spectral shifts observed at low and high temperatures. At low tempere-ture, Ty T and the center of gravity is Tb. At high 

Ufedme-resolved onission spectra of TNS-labeled DMPC/cholesterol vesicles are shown in Hgure 7.20. In 

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This chapter presents new information about the physical properties of humic acid fractions from the Okefenokee Swamp, Georgia. Specialized techniques of fluorescence depolarization spectroscopy and phase-shift fluorometry allow the nondestructive determination of molar volume and shape in aqueous solutions. The techniques also provide sufficient data to make a reliable estimate of the number of different fluorophores in the molecule their respective excitation and emission spectra, and their phase-resolved emission spectra. These measurements are possible even in instances where two fluorophores have nearly identical emission specta. The general theoretical background of each method is presented first, followed by the specific results of our measurements. Parts of the theoretical treatment of depolarization and phase-shift fluorometry given here are more fully expanded upon in (5,9-ll). Recent work and reviews of these techniques are given by Warner and McGown (72). 

Figures 3a-f show the emission and excitation spectra for all six humic fractions. The excitation and emission maxima are listed in Table III along with the maxima of the phase-resolved emission spectra. In each case the emission spectrum was scanned with the excitation maximum wavelength held constant, and the excitation spectrum was scanned with the emission maximum wavelength held constant. Several interesting features are noted. The two humic samples ( Figures 3a,b) each have two excitation maxima and it appears that a double peak has been merged into the emission scan as evidenced by the shoulder on the high energy side of the emission peak. Similarly it seems evident that the exaggerated shoulders in the emission spectra of all the fractions point to the inclusion of two emission peaks in each spectrum. This evidence suggests the presence of two chromophores in each humic fraction.

Jaye AA, Stoner-Ma D, Matousek P, Towrie M, Tonge PJ, Meech SR (2005) Time resolved emission spectra of green fluorescent protein. Photochem Photobiol 82 373-379... 

Emission spectra of radical cations are obtained by vacuum UV ionization and subsequent laser excitation in noble-gas matrices (see below), or by electron-impact ionization of a beam of neutral parent molecules at energies above the first ionic excited state. After internal conversion to the first excited state, emission may compete more or less successfully with radiationless deactivation. If the experiment is carried out on a supersonic molecular beam one obtains highly resolved emission spectra which, in the case of small molecules, may contain sufficient information to allow a determination of the molecular structure. 

As an example for linear x2-fitting, we analyse time resolved emission spectra of a mixture of two components with overlapping emission spectra and similar lifetimes. 

Among other examples, time-resolved luminescence has recently been applied to the detection of different trace elements (i.e., elements in very low concentrations) in minerals. Figure 1.13 shows two time-resolved emission spectra of anhydrite (CaS04). The emission spectrum just after the excitation pulse (delay 0 ms) shows an emission band peaking at 385 nm, characteristic of Eu + ions. When the emission spectmm is taken 4 ms after the pulse, the Eu + luminescence has completely disappeared, as this luminescence has a lifetime of about 10/rs. This allows us to observe the weak emission signals of the Eu + and Sm + ions present in this mineral, which in short time intervals are masked by the En + Inminescence. The trivalent ions have larger lifetimes and their luminescence still remains in the ms delay range. 

The narrow band with two maxims at 335 and 360 nm in time-resolved emission spectra of datolite (Fig. 4.16a), with a short decay time of 30 ns is connected with Ce +. 

The time-resolved emission spectra (TRES) and fluorescence lifetimes, ti, of the fluorene derivatives were measured in liquid solutions at room temperature with a PTI QuantaMaster spectrofluorimeter with 0.1 ns temporal resolution [20]. At this resolution, all investigated fluorenes exhibited TRES which were coincident with the corresponding steady-state fluorescence spectra. As an example, TRES for compounds 3 and 11 in hexane, THE, and ACN are presented in Eig. 8 for different nanosecond delays 0 ns (curves 2,4,6) and 5 ns, which modeled the steady-state condition (curves 3,5,7). No differences in the fluorescence spectra for these two delays were observed, indicating that all relaxation processes in the first excited state Si are sufficiently fast for fluorene molecifles and did not exceed the time resolution of the PTI system ( 0.1 ns). 

Time-resolved emission spectra are reconstructed from the normalized impulse response functions (26) ... 

Once the time-resolved emission spectra are so generated, it is informative to monitor the time evolution of the emission spectra. To this end, it is convenient to focus on the time course of the emission center of gravity (i>(t)) (27) ... 

Time-resolved emission spectra were reconstructed from a set of multifrequency phase and modulation traces acquired across the emission spectrum (37). The multifrequency phase and modulation data were modeled with the help of a commercially available global analysis software package (Globals Unlimited). The model which offered the best fits to the data with the least number of fitting parameters was a series of bi-exponential decays in which the individual fluorescence lifetimes were linked across the emission spectrum and the pre-exponential terms were allowed to vary. 

The time-resolved emission spectra were reconstructed from the fluorescence decay kinetics at a series of emission wavelengths, and the steady-state emission spectrum as described in the Theory section (37). Figure 4 shows a typical set of time-resolved emission spectra for PRODAN in a binary supercritical fluid composed of CO2 and 1.57 mol% CH3OH (T = 45 °C P = 81.4 bar). Clearly, the emission spectrum red shifts following excitation indicating that the local solvent environment is becoming more polar during the excited-state lifetime. We attribute this red shift to the reorientation of cosolvent molecules about excited-state PRODAN. 

Since exact formulas for time and frequency resolved emission spectra are less standard we shortly comment on the derivation of the full quantum expressions [43-45]. To characterize the emission we introduce the rate R k(t) which follows as the number of photons emitted per time into the state with polarization A and wave vector k. Since emission appears into the photon vacuum we may set R k = dN k/dt, where... 

Steady-state fluorescence spectra recorded after the addition of the NDI or PM I acceptor to the bisporphyrin tweezer ( rrl = 660 nm), demonstrated substantial quenching (75%) with increasing quantities of the NDI or PM I acceptors. Time-resolved emission spectra recorded in toluene for the complex 26 were biexponential containing a dominant short-lived CS components (80 ps, -95%) attributed to photoinduced ET from donor porphyrin to NDI, and a minor long-lived component (Ins, 5%). The lifetime of the dominant short-lived CS state is increased two- to threefold relative to covalently linked systems under similar conditions of solvent, donor-acceptor distance and thermodynamics [37]. Charge recombination rates from 1.4 to 3.8 x 1()9s 1 were observed, depending on whether the NDI or PM I acceptor was bound within the cavity. 

Fig. 22 Emission spectra of polycrystalline [AuI C(NHMe)2 2](PF6)-0.5(acetone). A Emission (A,excitation 383 nm) and excitation (Aemission 482 nm) spectra at 300 K. B Time resolved emission spectra at 300 K solid line emission acquired within 50 ns of a 337 nm laser pulse dotted line emission acquired 200 ns after the laser pulse at 295 K. C Emission (Aexcitation 365 nm) spectrum at 77 K. From [48]...

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) ... 

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