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By Edwin S. Crawley, Henry B. Evans

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In our instrumentation the count rates become significantly nonlinear above 100,000 photons per sec. Photon-counting detection is therefore inconvenient when signal levels are high. To stay within the linear range, one must adjust the slit widths or the fluorescence intensities (using neutral density filters). OOQphotons per second. This limited intensity range is a drawback of photon-counting detection for steady state measurements of fluorescence. In the analogue mode the individual pulses are averaged.

2 Pen-Ray Lamp from Ultra-Violet Products, San Gabriel, California. The values of the relative intensities are only approximate. intensity. It is important to attenuate the light so that the photomultiplier tube and/or amplifiers are not damaged. Following these precautions, one locates the dominant Hg lines using the emission monochromator. 1. If the observed values differ from these values by a constant amount one recalibrates the monochromator to obtain coincidence. 1 by an amount which is wavelength dependent.

This is because only a fraction of the sample is illuminated and the bleached portion of the sample is continuously replaced by fresh solution. Grating monochromators may have planar or concave gratings. Planar gratings are usually produced mechanically and may contain imperfections in some of the grooves. Concave gratings are usually produced by holographic and photoresist methods, and imperfections are rare. Imperfections of the gratings are the major source of stray light transmission by the monochromators, and of ghost images from the grating.

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