Time-resolved optical polarization

The lineshape of the VII spectrum s dominant mode was obtained by Koch, et al. (4) for 23 and 48 kDa polystyrenes in cyclohexane at concentrations 690-810 g/l. Their field correlation functions were accurately described by a Williams-Watts function exp(—(t/r) ). For a 700 g/l solution, = 0.4 was independent of temperature for 21 T 52°C. The mean relaxation time (tv//) = for depolarized scattering had the Vogel-Fulcher-Tamman temperature dependence 

In a time-resolved fluorescence experiment, a very short (ps) polarized light pulse is used to excite chromophores in the sample, the probability that a chromophore is excited being determined by the angle at time of absorption between the polarization vector of the light and the transition dipole of the chromophore. Each excited chromophore re-emits fluorescently. The polarization of each fluorescent photon is determined by the orientation of the transition dipole, at time of emission, as seen from the detector. The key variable is the time interval t between absorption and emission. At t = 0, the chromophore transition dipole has its initial orientation. As t increases, rotational diffusion leads to reorientation of the transition dipole. Correspondingly, at t = 0 the polarization of fluorescent emission is maximally correlated with the polarization of the exciting light as t increases, the fluorescent emission is less and less correlated with the initial polarization. 

Mathematical complications, discussed in the original papers, appear because the transition dipoles initially have random orientations in space relative to the polarization of the exciting pulse. In a representative experiment, the incident pulse is polarized perpendicular to the scattering plane, and the time-dependent intensities 

Considering first experiments in which the fluorophore is at the exact center of the polymer chain, with its transition dipole covalently bonded to lie along the chain axis, there are studies in which the solvent is changed, in which pressure and temperature are primary variables, and in which nondilute solutions are examined. Limited work treats polymers in which the fluorophore is not centrally located in the chain. 

An alternative experimental approach is presented by Hyde, et al.(S), who examined reorientation via holographic grating methods. In these experiments, picosecond polarized laser pulses passing through the sample along two nonparallel 

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Photoderacemization is the simplest case of direct asymmetric photoreactions induced by cpl. The enantiomers are interconverted, and the mixture becomes optically active. Reaction scheme 2 is a modification of Scheme 1 ground state racemization is excluded. The enantiomerization step R S was observed directly by Metcalf et al. [9] by means of the time-resolved circularly polarized luminescence of europium-tris(bipicolinate). By means of a cpl laser pulse, a difference in the excited state population is created, and the decay of circular... 

Time-resolved optical experiments rely on a short pulse of polarized light from a laser, synchrotron, or flash lamp to photoselect chromophores which have their transition dipoles oriented in the same direction as the polarization of the exciting light. This non-random orientational distribution of excited state transition dipoles will randomize in time due to motions of the polymer chains to which the chromophores are attached. The precise manner in which the oriented distribution randomizes depends upon the detailed character of the molecular motions taking place and is described by the orientation autocorrelation function. This randomization of the orientational distribution can be observed either through time-resolved polarized fluorescence (as in fluorescence anisotropy decay experiments) or through time-resolved polarized absorption. 

The spin polarization of the anteima carotenoid triplet state has been observed by Frank et al. (1980 1982a 1987) in quite a number of different purple bacterial strains, and under all conditions shows an eae aea pattern (where e means emission and a absorption of microwaves) that can be explained with intersystem crossing in a BChl molecule with subsequent triplet energy transfer to the carotenoid. This seems to contradict the additional triplet formation pathway by hetero-fission of a carotenoid singlet excitation and it would therefore be of great interest to revisit the earlier time-resolved optical... 

Experimental work is consistent with the view that solvent specific corrections to the intramolecular potential are not required in the absence of strong polymer/solvent interactions. Glowinkowski et al. used CNMR to study the local dynamics of polyisoprene in ten solvents as a function of temperature [31]. They measured correlation times due to the motion of differmt C-H vectors in the chain backbone. They found that these correlation times were determined by oidy the temperature and the solvent viscosity. Variables such as solvent shape, flexibility, and chemical functionality had no effect on the correlation times, except through the solvent viscosity. (Highly polar solvents were excluded from this study as they do not dissolve polyisoprene.) Similar conclusions have been reached in NMR studies of polybutadiene [52] and in time-resolved optical studies of polyisoprene [53] and polystyrene [54] with anthracene labels. NMR studies of PEO [55] have been interpreted as supporting this same conclusion [31] (except when the solvent was water [56]). 

An interesting feature of polarized IR spectroscopy is that rapid measurements can be performed while preserving molecular information (in contrast with birefringence) and without the need for a synchrotron source (X-ray diffraction). Time-resolved IRLD studies are almost exclusively realized in transmission because of its compatibility with various types of tensile testing devices. In the simplest implementation, p- and s-polarized spectra are sequentially acquired while the sample is deformed and/or relaxing. The time resolution is generally limited to several seconds per spectrum by the acquisition time of two spectra and by the speed at which the polarizer can be rotated. Siesler et al. have used such a rheo-optical technique to study the dynamics of multiple polymers and copolymers [40]. 

By using a nonlinear optical process such as SHG, one can probe surface phonons and adsorbate-related vibrations exclusively [14,15,32,34]. Time-resolved SHG (TRSHG) detects the second harmonic (SH) of the probe beam as a function of time delay between pump and probe. The SH electric field is driven by the nonlinear polarization Pi 2w) at the surface, which... 

Fig. 3.5. Experimental apparatus for time-resolved THz transmission spectroscopy. The sample is excited with a visible laser pulse delivered by delay line 3. A singlecycle THz electric-field transient probes the polarization response of the sample after time delay tv scanned by delay line 1. The transmitted THz amplitude is monitored via ultrabroadband electro-optic sampling in a THz receiver as a function of time T scanned by delay line 2. From [13]...

Figure 4.1. Time scales for rotational motions of long DNAs that contribute to the relaxation of the optical anisotropy r(t). Experimental methods used to study these motions in different time ranges are also indicated along with the authors and dates of some early work in each case. FPA, Fluorescence polarization anisotropy (Refs. 15, 18-20, and 87) TPD, transient photodichroism (Refs. 28 and 62) TEB, transient electric birefringence (Refs. 26 and 27) DDLS, depolarized dynamic light scattering (Ref. 116) TED, transient electric dichroism (Refs. 25, 115, and 130) Microscopy, time-resolved fluorescent microscopy (Ref. 176).

At the present time, two methods are in common use for the determination of time-resolved anisotropy parameters—the single-photon counting or pulse method 55-56 and the frequency-domain or phase fluorometric methods. 57 59) These are described elsewhere in this series. Recently, both of these techniques have undergone considerable development, and there are a number of commercially available instruments which include analysis software. The question of which technique would be better for the study of membranes is therefore difficult to answer. Certainly, however, the multifrequency phase instruments are now fully comparable with the time-domain instruments, a situation which was not the case only a few years ago. Time-resolved measurements are generally rather more difficult to perform and may take considerably longer than the steady-state anisotropy measurements, and this should be borne in mind when samples are unstable or if information of kinetics is required. It is therefore important to evaluate the need to take such measurements in studies of membranes. Steady-state instruments are of course much less expensive, and considerable information can be extracted, although polarization optics are not usually supplied as standard. 

Probing Metalloproteins Electronic absorption spectroscopy of copper proteins, 226, 1 electronic absorption spectroscopy of nonheme iron proteins, 226, 33 cobalt as probe and label of proteins, 226, 52 biochemical and spectroscopic probes of mercury(ii) coordination environments in proteins, 226, 71 low-temperature optical spectroscopy metalloprotein structure and dynamics, 226, 97 nanosecond transient absorption spectroscopy, 226, 119 nanosecond time-resolved absorption and polarization dichroism spectroscopies, 226, 147 real-time spectroscopic techniques for probing conformational dynamics of heme proteins, 226, 177 variable-temperature magnetic circular dichroism, 226, 199 linear dichroism, 226, 232 infrared spectroscopy, 226, 259 Fourier transform infrared spectroscopy, 226, 289 infrared circular dichroism, 226, 306 Raman and resonance Raman spectroscopy, 226, 319 protein structure from ultraviolet resonance Raman spectroscopy, 226, 374 single-crystal micro-Raman spectroscopy, 226, 397 nanosecond time-resolved resonance Raman spectroscopy, 226, 409 techniques for obtaining resonance Raman spectra of metalloproteins, 226, 431 Raman optical activity, 226, 470 surface-enhanced resonance Raman scattering, 226, 482 luminescence... 

The optical setup for time-resolved micro-luminescence measurements is based around an Axiotech 100 HD Zeiss microscope, modified to allow laser injection and fluorescence collection. The sample is observed either under transmission or reflection of polarized white light, or under UV illumination (HBO lamp). A set-up consisting of a dichroic mirror, for the selection of the excitation wavelength, and an objective (Epiplan Neofiuar obj. > 350 nm Ealing/Coherent reflection obj. < 350 nm) is used to focus the laser beam on the sample (spatial resolution over 5 pm with a x50 objective). The lim-... 

This mechanism leads to a highly spin-polarized triplet state with a characteristic intensity pattern in the EPR spectrum, which is observed by time-resolved techniques (either transient or pulse EPR). The zero field splitting (ZFS) of the triplet state, which dominates the EPR spectrum, is an important additional spectroscopic probe. It can also be determined by optical detection of magnetic resonance (ODMR), for a review of the techniques involved and applications see reference 15. These methods also yield information about dynamical aspects related to the formation, selective population and decay of the triplet states. The application of EPR and related techniques to triplet states in photosynthesis have been reviewed by several authors in the past15 22-100 102. The field was also thoroughly reviewed by Mobius103 and Weber45 in this series. 

Further work using time-resolved EPR and magnetophotoselection (MPS), using plane-polarized light to excite the triplet state, gave information on the orientation of the optical transition dipole axes relative to the principal axes of the triplet state. By this technique the transition moments of the primary donor"6, the carotenoid in the bRC"7 and the bacteriopheophytin in the inactive B branch 4>0"8 were determined. 

Time-resolved fluorescence spectroscopy of polar fluorescent probes that have a dipole moment that depends upon electronic state has recently been used extensively to study microscopic solvation dynamics of a broad range of solvents. Section II of this paper deals with the subject in detail. The basic concept is outlined in Figure 1, which shows the dependence of the nonequilibrium free energies (Fg and Fe) of solvated ground state and electronically excited probes, respecitvely, as a function of a generalized solvent coordinate. Optical excitation (vertical) of an equilibrated ground state probe produces a nonequilibrium configuration of the solvent about the excited state of the probe. Subsequent relaxation is accompanied by a time-dependent fluorescence spectral shift toward lower frequencies, which can be monitored and analyzed to quantify the dynamics of solvation via the empirical solvation dynamics function C(t), which is defined by Eq. (1). 

Time-Resolved Chemically Induced Dynamic Electron Polarization and Optical Emission Studies... 

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