Spectral sensitivity, measurement

Figure 3. Optical system of instrument of spectral sensitivity measurement (L) Xe arc lamp (Ml) concave mirror (M2, M3) mirror (S) shutter (SI) slit (G) concave reflective grating...

Spectroscopic methods such as uv and fluorescence have rehed on the polyene chromophore of vitamin A as a basis for analysis. Indirectly, the classical Carr-Price colorimetric test also exploits this feature and measures the amount of a transient blue complex at 620 nm which is formed when vitamin A is dehydrated in the presence of Lewis acids. For uv measurements of retinol, retinyl acetate, and retinyl palmitate, analysis is done at 325 nm. More sensitive measurements can be obtained by fluorescence. Excitation is done at 325 nm and emission at 470 nm. Although useful, all of these methods suffer from the fact that the method is not specific and any compound which has spectral characteristics similar to vitamin A will assay like the vitamin... 

Lee, J., and Seliger, H. H. (1965). Absolute spectral sensitivity of phototubes and the application to the measurement of the absolute quantum yields of chemiluminescence and bioluminescence. Photochem. Photobiol. 4 1015-1048. 

Any color signal received from the snbject will be rated by the spectral sensitivity of the eye. The physiological bases for this are three types of cone-shaped receptors on the human retina that are sensitive to red, green, and bine. The color measurement techniques have been established on this tri-colorimetric system. They simulate human perception by the relative extent to which the observer estimates the relative share of which of these three components contributes to his color impression from an object. 

The cancellation of gas phase spectral features using the "half plate design Is far superior to methods Involving a second gas cell placed In the reference beam. This Is because the gas density and Its rotational state population will differ In the two cells for different sample (and therefore gas) temperatures. For high sensitivity measurements, these effects can be difficult to handle using two cells. 

Emission from dimols of singlet oxygen may be detected by photomultipliers used for measurement of chemiluminescence from hydrocarbon polymers with a maximum spectral sensitivity at 460 nm. The above scheme, however, requires the presence of at least one molecule of hydrogen peroxide in close vicinity to the two recombining peroxyl radicals and assumes a large heterogeneity of the oxidation process. 

The spectral characteristics of Lucifer Yellow iodoacetamide produce luminescence at somewhat higher wavelengths than the green luminescence of fluorescein, thus the yellow designation in its name. The excitation maximum for the probe occurs at 426 nm and its emission at 530 nm. The rather large Stoke s shift makes sensitive measurements of emission intensity possible without interference by scattered excitation light. The 2-mercaptoethanol derivative of the fluorophore has an extinction coefficient at pH 7 of about 13,000 M cm-1 at 426nm. 

Programming a CAM for fluorometry is far more complex than for spectrophotometry. Spectrophotometry is simple because it is based on the ratio of light in to light out. But fluorometry creates many of the problems associated with true radiometry—measuring the emission spectrum of an unknown source. The logic may become circular. Radiometry to determine an emission spectrum requires the relative spectral sensitivity of the photometer to be known, but how can this be determined without a source with a known emission spectrum Fortunately, physicists in our national standardization organizations provide us with calibrated sources and photometers. 

Because sensitivity depends on so many different experimental factors, NMR spectroscopists generally use the signal-to-noise ratio, SIN, as a figure of merit for sensitivity comparisons. For example, in a comparison between NMR probes or spectrometers from two vendors, the spectral SIN measured for a standard sample acquired with specified acquisition parameters and probe geometry would provide a direct indication of relative sensitivity. The SIN is calculated for an NMR experiment as the peak signal divided by the root mean square (RMS) noise, given by Equation 7.6, and is directly related to the performance of the radiofrequency coil [3,6]... 

A fluorescence emission spectrum is generally measured by setting the excitation monochromator, Mi, to the chosen wavelength and scanning the second monochromator, M2, with constant slit width. The fluorescent screen monitor, F-P2, now serves to correct for variations in the intensity of the exciting light caused by fluctuations in lamp output. The emission spectrum so recorded has to be corrected for the spectral sensitivity of the apparatus to give the true emission spectrum. 

The method just described is not usually applicable in the ultraviolet because ultraviolet lamps of known spectral distribution are not readily available at present. The spectral sensitivity caii be calculated directly if the values of B L and P, are known. The first of these is obtained from the dispersion curve of the monochromator the second is somewhat difficult to measure—for prism instruments over restricted wavelength regions above 250 m t it is often reasonably constant. The photomultiplier sensitivity, P can be determined by comparison with a thermopile or with the ferrioxalate actinometer.11 12 Direct calculation of S, is subject to inaccuracies due to the accumulation of errors in the measurement of the three separate quantities B L and P,. A more convenient... 

If one of the substances has a known fluorescence efficiency, the value of the other is then simply obtained. Convenient standard solutions are rhodamine B in ethanol with fluorescence in the yellow and efficiency 0.69, quinine bisulfate in 0.1 N sulfuric acid with fluorescence in the blue and efficiency 0.55. anthracene in ethanol with fluorescence in the violet and efficiency 0.27 in the ultraviolet region, naphthalene ( = 0.19), phenol (0 = 0.19), or benzene (0 = 0.042) can be used. With the last four compounds the solution must be deaerated by passing a current of nitrogen before measurement. To minimize the effect of errors in the spectral sensitivity curve it is desirable to use as the standard a solution... 

As a cross-check of this result a spectral simulation with a PT of prolate symmetry is given in Fig. 4c, The difference to the other traces is seen in the missing K-type recurrences. The occurrence of K-type transients in fs DFWM spectra can be rationalized by significant components of the polarizability tensor perpendicular to the symmetric top axis. Finally, it was demonstrated that the types and intensities of recurrences in fs DFWM spectra are a sensitive measure for the symmetry of the PT of the corresponding molecule. 

The applicability of this in situ method for the determination of surface areas depends not only on knowledge of the dye s molecular area in the adsorbed state but also on the assumption that the chosen spectral parameter measures the surface concentration of the dye. In order to test the relation between adsorption of dye to silver halide and its spectral characteristics in the bound state, the behavior of Pseudocyanine in a coarse silver halide suspension (Dispersion D) was studied. This particular dispersion was chosen because some of its relevant adsorption characteristics had already been examined (22, 23). Moreover, observations by Boyer and Cappelaere with Pseudocyanine adsorbed on AgBr powders (5) indicated that /-band intensity varied with the amount of adsorbed dye and was not sensitive to the concentration of Ag+ or Br" ions in the range pAg 3.3-8.7. 

This calibration factor can be determined at any instant from (1), if the spectrum of solar radiation (from the sun and sky) can be measured in absolute units and if the detector s relative spectral sensitivity w(X) is known. 

For the determination of the spectral sensitivity complicated and expensive laboratory equipment are required in conjunction with experienced personnel. Such a facility should be capable in providing calibrated monochromatic radiation of sufficient power to be sensed by the broadband detector in question. A typical system should be comprised by a double monochromator coupled with a stable, high-power source (e.g. a 1000-Watt Xenon lamp). A spectrally calibrated detector should also be available to measure the quasi-monochromatic radiation emitted by this system. Such facilities are rarely available and only a few laboratories worldwide are able to cany out such characterizations. 

Muenter and Cooper (30) measured the room-temperature fluorescence of two J-aggregated dyes, l,l -diethyl-2,2 -quino-cyanine and 1,1, 3,3 -tetraethyl-5,5, 6,6 -tetrachlorobenzimid-azolocarbocyanine, adsorbed on cubic AgBr grains. The quantum efficiency of spectral sensitization was inversely related to the relative fluorescence. The fluorescence by the dyes in the molecular state was low compared to that for the aggregated dyes. Addition of a styryl or thiohydantoin dye as a supersensitizer for the quinocyanine quenched its fluorescence and increased the relative efficiency from 0.06 to nearly 1.0. 

The effect of moisture alone on spectral sensitization has been studied to only a limited extent. Moist oxygen generally causes greater desensitization than dry oxygen, just as in the undyed emulsions. However, moisture alone can cause effects in some dyed emulsions that are not found in the undyed emulsions. Absorptance measurements made on several emulsions showed that vacuum outgassing decreased the absorptance in the region of the... 

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