Emission background

Minimizing Spectral Interferences The most important spectral interference is a continuous source of background emission from the flame or plasma and emission bands from molecular species. This background emission is particularly severe for flames in which the temperature is insufficient to break down refractory compounds, such as oxides and hydroxides. Background corrections for flame emission are made by scanning over the emission line and drawing a baseline (Figure 10.51). Because the temperature of a plasma is... 

As httle as lO " g of ATP can be detected with carefiiUy purified luciferase. Commercial luciferase contains enough residual ATP to cause background emission and increase the detection limit to 10 g (294). The method has been used to determine bacterial concentrations in water. As few as lO" cells/mL of Lscherichia coli, which contains as Httle as 10 g of ATP per cell, can be detected (294). Numerous species of bacteria have been studied using this technique (293—295). 

Typical elemental detection limits are listed in Table 1. The detection limit is the concentration that produces the smallest signal that can be distinguished from background emission fluctuations. The continuum background is produced via radiative recombination of electrons and ions e — M+ hv or M + e + e — ... 

Direct-reading polychromators (Figure 3b) have a number of exit slits and photomultiplier tube detectors, which allows one to view emission from many lines simultaneously. More than 40 elements can be determined in less than one minute. The choice of emission lines in the polychromator must be made before the instrument is purchased. The polychromator can be used to monitor transient signals (if the appropriate electronics and software are available) because unlike slew-scan systems it can be set stably to the peak emission wavelength. Background emission cannot be measured simultaneously at a wavelength close to the line for each element of interest. For maximum speed and flexibility both a direct-reading polychromator and a slew-scan monochromator can be used to view emission from the plasma simultaneously. 

An additional limit to the size of a passive array relates to the current which flows in an OLED when it is under reverse bias [189]. When a given pixel is turned on in the array, there are many possible parallel paths for the current, each involving two diodes in reverse bias and one forward. Hence, as the number of rows and columns increases, there is a higher level of background emission from non-selected pixels that limits the contrast ratio of the array. As a result, the contrast degrades as N increases. 

Capomacchia et al. [120] utilized the background emission present when DNPO and hydrogen peroxide are mixed to detect ouabain and urea. In the presence of these analytes, an intensity enhancement was observed and detection limits were in the picomole range. 

Increased sensitivity by virtue of the high hydrogen peroxide-to-aryl oxalate ratios used, which facilitate suppression of background emission and thus raise the signal-to-noise ratios. 

Versatility Another benefit that derives from the fact that CL reagents are continuously mixed in front of the detector, regardless of the presence of the analyte, is implementation of the analytical procedure even in cases of reaction where the reagents produce a low background emission. This happens because this background emission can be regarded as the baseline since it is constant with the time and, hence, it will not interfere with the analytical signal produced by the analyte. 

A background emission underlies all chemiluminescence spectra obtained in reactions of O atoms. This emission results from the recombination of O atoms,... 

Emissions from five different excited electronic states of 02 throughout the visible spectral region of 400-800 nm have been identified [17], placing a practical limit on the concentration of O atoms that can be used as a chemiluminescence reagent since the background emission increases quadratically in O atom concentration. 

Background emission by the flame (Figure 8.23) includes contributions from molecular species and continuum radiation from incandescent particles and depends upon the combination of fuel and support gases used. The sample solvent and matrix will further augment background radiation. 

Spectral interferences may arise from the close proximity of other emission lines or bands to the analyte line or by overlap with it. They can often be eliminated or minimized by increasing the resolution of the instrumentation, e.g. changing from a filter photometer to a grating spectrophotometer. Alternatively, another analyte line can be selected for measurements. Correction for background emission is also important and is made by monitoring the emission from a blank solution at the wavelength of the analyte line or by averaging measurements made close to the line and on either side of it. 

The procedure is strictly analogous to that used for absorbance measurements in UV and visible molecular spectrometry (p. 355). To avoid interference from emission by excited atoms in the flame and from random background emission by the flame, the output of the lamp is modulated, usually at 50 Hz, and the detection system tuned to the same frequency. Alternatively, a mechanical chopper which physically interrupts the radiation beam, can be used to simulate modulation of the lamp output. 

The sample is continuously irradiated and the fluctuations in the fluorescence intensity arise due to any event which makes the fluorophore unavailable to be excited to the emissive singlet excited state, such as diffusion of the fluorophore out of the detection volume, formation of a dark state, such as a triplet excited state, or photoreaction. The concentration of fluorophore in the detection volume has to be low (10 13—10 8M) so that the fluctuation in the intensity for one molecule is observable over any background emission. The high concentration limit is a consequence of the fact that the correlated photons from single molecules scale with the number of molecules in the detection volume, while the contribution from uncorrelated photons, arising from the emission from different molecules, scales with the square of the number of molecules. The lowest concentration is determined by the probability of finding a molecule in the detection volume.58... 

The autocorrelation function G(t) corresponds to the correlation of a time-shifted replica of itself at various time-shifts (t) (Equation (7)).58,65 This autocorrelation defines the probability of the detection of a photon from the same molecule at time zero and at time x. Loss of this correlation indicates that this one molecule is not available for excitation, either because it diffused out of the detection volume or it is in a dark state different from its ground state. Two photons originating from uncorrelated background emission, such as Raman scattering, or emission from two different molecules do not have a time correlation and for this reason appear as a time-independent constant offset for G(r).58... 

In addition to the emission due to the test element, radiation is also emitted by the flame itself. This background emission, together with turbulence in the flame, results in fluctuations of the signal and prevents the use of very sensitive detectors. The problem may be appreciably reduced by the introduction into the sample of a constant amount of a reference element and the use of a dual-channel flame photometer, which is capable of recording both the test and reference readings simultaneously. The ratio of the intensity of emission of the test element to that of the reference element should be unaffected by flame fluctuations and a calibration line using this ratio for different concentrations of the test element is the basis of the quantitative method. Lithium salts are frequently used as the reference element in the analysis of biological samples. 

Improved LIF sensing discrimination power is required for sample matrices that contain multiple fluorophores with similar spectral emission properties or when background emission is problematic. Distinguishing among airborne bioagent hazards and common emissive interferants (albuminous, epithelium, and cellulous materials as well as aromatic hydrocarbons), is a prime example where higher selective detection is required. This can be achieved via the lifetime properties of each fluorophore, by an optode approach or both. 

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