Resonance Raman methods

By resonance Raman methods, Wang and coworkers105 showed NO bound to both ferric and ferrous heme of bNOS. Hurshman and Marietta106 used iNOS and spectrophotometric methods to show similar reactions, although the physiological effects will depend on the effects of arginine and oxygen in vivo. 

Because of the high sensitivity required, this generally involves the measurement of intense electronic transitions. In principle, infrared or Raman detection could be more widely applicable, but, except for resonance Raman methods, which again depend on the presence of UV-visible bands, these methods are too insensitive to be of much use at present. 

Infrared and Resonance Raman Spectroscopy. Reviewson the uses of resonance Raman spectroscopy in biochemistry and biology include sections on carotenoproteins, visual pigments, and bacteriorhodopsin. The resonance Raman spectrum of the lowest excited triplet state of /3-carotene has been reported.A resonance Raman method has been used for the quantitative analysis of /3-carotene and lutein (20) in tobacco.The mechanism of carotenoid-protein interactions in the carotenoproteins ovoverdin and /3-crustacyanin has been investigated by resonance Raman spectroscopy. " 2 axanthin (24) has been used as a resonance Raman probe of membrane structure. " The resonance Raman spectra have been reported of all-frans-anhydrovitamin A (194), " /3-ionone, retinals, and Schiff bases.The technique has been used extensively to study... 

Thamann, Loehr and Loehr73) continued to examine the 0—0 stretch using unsymmetrically labelled 02 (1602 + 1802 + 160 180) by the resonance Raman method. They found three Raman peaks at 749,728 and 708 cm-1 for the labelled molluscan oxyhemocyanin and the peak intensities were the same as observed for the 02 gas mixture used. That requires that both oxygen atoms be equivalent in the bound form and strongly supports a bridging peroxide between the two Cu (II) ions. A terminal symmetrical peroxide (a) would be expected to have an... 

Porphyrins absorb strongly in the visible spectral regions and are therefore ideal candidates for study using resonance Raman methods. 

Resonance enhancement can be used to remove fluorescence by making the Raman signal much more intense than the fluorescence signal. The UV-resonance Raman method can be used to enhance the Raman signal above the fluorescence signal if the sample has the proper electronic structure for resonance enhancement and if the proper UV frequency source is available. The resonance Raman technique is discussed in the section Resonance Raman Spectroscopy (p. 229) in this chapter. 

The siim-over-states method for calculating the resonant enlrancement begins with an expression for the resonance Raman intensity, /.y, for the transition from initial state to final state /in the ground electronic state, and is given by [14]... 

It is also possible to detennine the resonant Raman intensities via a time-dependent method [16]. It has the... 

The advantages of resonance Raman spectroscopy have already been discussed in section BL2.2.3. For these reasons it is rapidly becoming the method of choice for studying large molecules in solution. Flere we will present one study that exemplifies its attributes. There are two complementary methods for studying proteins. 

Kincaid J R 1995 Structure and dynamics of transient species using time-resolved resonance Raman-spectroscopy Biochemical Spectroscopy Methods Enzymol. vol 246, ed K Sauer (San Diego, CA Academic) pp 460-501... 

Hizhnyakov V and Tehver I 1988 Transform method in resonance Raman scattering with quadratic Franck-Condon and Herzberg-Teller interactions J. Raman Spectrosc. 19 383-8... 

Friedman J M 1994 Time-resolved resonance Raman spectroscopy as probe of structure, dynamics, and reactivity in hemoglobin Methods Enzymol. 232 205-31... 

In an ambitious study, the AIMS method was used to calculate the absorption and resonance Raman spectra of ethylene [221]. In this, sets starting with 10 functions were calculated. To cope with the huge resources required for these calculations the code was parallelized. The spectra, obtained from the autocorrelation function, compare well with the experimental ones. It was also found that the non-adiabatic processes described above do not influence the spectra, as their profiles are formed in the time before the packet reaches the intersection, that is, the observed dynamic is dominated by the torsional motion. Calculations using the Condon approximation were also compared to calculations implicitly including the transition dipole, and little difference was seen. 

Band gaps in semiconductors can be investigated by other optical methods, such as photoluminescence, cathodoluminescence, photoluminescence excitation spectroscopy, absorption, spectral ellipsometry, photocurrent spectroscopy, and resonant Raman spectroscopy. Photoluminescence and cathodoluminescence involve an emission process and hence can be used to evaluate only features near the fundamental band gap. The other methods are related to the absorption process or its derivative (resonant Raman scattering). Most of these methods require cryogenic temperatures. 

Ren et al. reported a method to prepare a gold tip with a tip apex radius of 30 nm reproducibly [27]. They observed the TERS of a Malachite Green isothiocyanate (MGITC) monolayer on an Au(lll) surface and obtained an enhancement factor of about 1.6 X 10, by using the relation, q= /TERs/lRRs=g /l focus where q is the net increase in the signal. Iters snd rrs the signal intensities for TERS and RRS (resonance Raman scattering), respectively is the TERS enhancement (gis the field enhancement), a denotes the radius of the enhanced field, and Rfocus the radius of the laser focus. 

Typical Instrumentation and Methods for Doing Time-Resolved Resonance Raman (TR ) Experiments... 

TYPICAL INSTRUMENTATION AND METHODS FOR DOING TIME-RESOLVED RESONANCE RAMAN (TR ) EXPERIMENTS... 

Since there are a large number of different experimental laser and detection systems that can be used for time-resolved resonance Raman experiments, we shall only focus our attention here on two common types of methods that are typically used to investigate chemical reactions. We shall first describe typical nanosecond TR spectroscopy instrumentation that can obtain spectra of intermediates from several nanoseconds to millisecond time scales by employing electronic control of the pnmp and probe laser systems to vary the time-delay between the pnmp and probe pnlses. We then describe typical ultrafast TR spectroscopy instrumentation that can be used to examine intermediates from the picosecond to several nanosecond time scales by controlling the optical path length difference between the pump and probe laser pulses. In some reaction systems, it is useful to utilize both types of laser systems to study the chemical reaction and intermediates of interest from the picosecond to the microsecond or millisecond time-scales. 

It can be seen from Figures 3.7 and 3.8 that the calculations reproduce very well not only the experimental spectra but also the experimentally observed isotopic shifts indicating a high reliability of the computational method. According to this comparison, definite attribution can be made for even the difficult Raman bands that cannot be assigned based solely on the experimental results. It is, however, necessary to mention at this point that the calculated Raman spectrum provided directly by the ab initio computations correspond to the normal Raman spectrum with the band intensity determined by the polarizability of the correlating vibration. Since the intensity pattern exhibited by the experimentally recorded resonance Raman spectrum is due to the resonance enhancement effect of a particular chromophore, with no consideration of this effect, the calculated intensity pattern may, in many... 

With recent developments in analytical instrumentation these criteria are being increasingly fulfilled by physicochemical spectroscopic approaches, often referred to as whole-organism fingerprinting methods.910 Such methods involve the concurrent measurement of large numbers of spectral characters that together reflect the overall cell composition. Examples of the most popular methods used in the 20th century include pyrolysis mass spectrometry (PyMS),11,12 Fourier transform-infrared spectrometry (FT-IR), and UV resonance Raman spectroscopy.16,17 The PyMS technique... 

When carotenoids such as lutein and zeaxanthin are excited by wavelengths in the -450-550 nm range, they exhibit particularly strong resonance Raman signals that can be used to quantify the amount of carotenoid present. The application of this technique for quantifying the macular carotenoids has been developed, thereby providing another noninvasive physical method for MP measurement. A detailed description of this method is given in Chapter 6. 

The identification of xanthophylls in vivo is a complex task and should be approached gradually with the increasing complexity of the sample. In the case of the antenna xanthophylls, the simplest sample is the isolated LHCII complex. Even here four xanthophylls are present, each having at least three major absorption transitions, 0-0, 0-1, and 0-2 (Figure 7.4). Heterogeneity in the xanthophyll environment and overlap with the chlorophyll absorption add additional complexity to the identification task. No single spectroscopic method seems suitable to resolve the overlapping spectra. However, the combination of two spectroscopic techniques, low-temperature absorption and resonance Raman spectroscopy, has proved to be fruitful (Ruban et al., 2001 Robert et al., 2004). 

---