Light emissions spectrum

Knowledge on the plasma species can be obtained by the use of plasma diagnostics techniques, such as optical emission spectroscopy (OES) and mass spectroscopy (MS). Both techniques are able to probe atomic and molecular, neutral or ionized species present in plasmas. OES is based on measuring the light emission spectrum that arises from the relaxation of plasma species in excited energy states. MS, on the other hand, is generally based on the measurement of mass spectra of ground state species. 

Figures. The light emission spectrum of the nanocomposite island gold film - Alqs under applied voltage of 22 V. The inset shows a schematic of the planar structure.

Luminescence reaction (Viviani et al., 2002a) The luciferin-luciferase luminescence reaction was carried out in 0.1 M Tris-HCl, pH 8.0, containing 2mM ATP and 4mM Mg2+. Mixing luciferase with luciferin and ATP resulted in an emission of light with rapid onset and a kinetically complex decay. Further additions of fresh luciferase, after the luminescence has decayed to about 10% of its maximum value, resulted in additional luminescence responses similar to the initial one (Fig. 1.15). According to the authors, the repetitive light emission occurred in consequence of the inhibition of luciferase by a reaction product, as seen in the case of the firefly system (McElroy et al., 1953). The luminescence spectrum showed a peak at 487nm (Fig. 1.16). 

The bioluminescence reaction of Oplophorus is a typical luciferin-luciferase reaction that requires only three components luciferin (coelenterazine), luciferase and molecular oxygen. The luminescence spectrum shows a peak at about 454nm (Fig. 3.3.1). The luminescence is significantly affected by pH, salt concentration, and temperature. A certain level of ionic strength (salt) is necessary for the activity of the luciferase. In the case of NaCl, at least 0.05-0.1 M of the salt is needed for a moderate rate of light emission, and about 0.5 M for the maximum light intensity. 

Harvey (1952) demonstrated the luciferin-luciferase reaction with O. phosphorea collected at Nanaimo, British Columbia, Canada, and with O. enopla from Bermuda. McElroy (1960) partially purified the luciferin, and found that the luminescence spectrum of the luciferin-luciferase reaction of O. enopla is identical to the fluorescence spectrum of the luciferin (A.max 510 nm), and also that the luciferin is auto-oxidized by molecular oxygen without light emission. Further investigation on the bioluminescence of Odontosyllis has been made by Shimomura etal. (1963d, 1964) and Trainor (1979). Although the phenomenon is well known, the chemical structure of the luciferin and the mechanism of the luminescence reaction have not been elucidated. 

Fig. 7.4.2 Luminescence spectrum of a solution of the photoprotein polynoidin. Light emission was initiated by the additions of H2O2 (final cone. 3mM) and Fe2+ (final cone. 0.1 mM). From Nicolas et al., 1982, with permission form the American Society for Photobiology.

The photoprotein is non-fluorescent. The absorption spectrum of purified photoprotein shows a very small peak at 410 nm, in addition to the protein peak at 280 nm (Fig. 10.2.5). The peak height at 410 nm appears to be proportional to the luminescence activity of the protein. The protein also shows extremely weak absorption peaks at about 497, 550 and 587nm (not shown). These absorption peaks, except the 280 nm peak, might be due to the presence of a chromophore that is functional in the light emission. 

Luminescence of Pyrosoma. All species of the genus Pyrosoma (about 10 species) are bioluminescent. Pyrosoma is one of the few organisms reported to luminesce in response to light (Bowlby et al., 1990). The luminescence emission spectrum of Pyrosoma atlantica is bimodal according to Kampa and Boden (1957), with the primary peak near 482 nm, and the secondary near 525 nm. Swift et al. (1977) reported the emission maxima of two Pyrosoma species at 485 and 493 nm, respectively, and Bowlby et al. (1990) found an emission peak at 475 nm with P. atlantica. A corrected bioluminescence spectrum of P. atlantica (A.max 485 nm) reported by Herring (1983) is shown in Fig. 10.5.2. 

If we pass white light through a vapor composed of the atoms of an element, we see its absorption spectrum, a series of dark lines on an otherwise continuous spectrum (Fig 1.11). The absorption lines have the same frequencies as the lines in the emission spectrum and suggest that an atom can absorb radiation only of those same frequencies. Absorption spectra are used by astronomers to identify elements in the outer layers of stars. 

Theory. If two or more fluorophores with different emission lifetimes contribute to the same broad, unresolved emission spectrum, their separate emission spectra often can be resolved by the technique of phase-resolved fluorometry. In this method the excitation light is modulated sinusoidally, usually in the radio-frequency range, and the emission is analyzed with a phase sensitive detector. The emission appears as a sinusoidally modulated signal, shifted in phase from the excitation modulation and partially demodulated by an amount dependent on the lifetime of the fluorophore excited state (5, Chapter 4). The detector phase can be adjusted to be exactly out-of-phase with the emission from any one fluorophore, so that the contribution to the total spectrum from that fluorophore is suppressed. For a sample with two fluorophores, suppressing the emission from one fluorophore leaves a spectrum caused only by the other, which then can be directly recorded. With more than two flurophores the problem is more complicated but a number of techniques for deconvoluting the complex emission curve have been developed making use of several modulation frequencies and measurement phase angles (79). 

Optical emision spectra nowadays are simply measured using a fiber optic cable that directs the plasma light to a monochromator, which is coupled to a photodetector. By rotating the prism in the monochromator a wavelength scan of the emitted light can be obtained. Alternatively, an optical multichannel analyzer can be used to record (parts of) an emission spectrum simultaneously, allowing for much faster acquisition. A spectrometer resolution of about 0.1 nm is needed to identify species. 

Procedure Set up an acoustic reactor in a light-proof cabinet with a photomultiplier (PM) tube positioned facing the cell as shown in Fig. 15.3a and b. Fill the cell with distilled water and close the cabinet. A potential should now be applied to the PM tube, the output (spectrally integrated) of which is produced on an oscilloscope (note that the ultrasound cell can easily be placed inside a commercial spectrometer in order to record the emission spectrum). Switch on the ultrasound and you should observe on the oscilloscope a change in voltage, directly proportional to the intensity of sonoluminescence emission. The following experiments can be performed to explore the different types of light emission and some of the factors that influence these emission processes. 

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