Wavelength Verification And Calibration - Agilent Technologies 1200 Series User Manual

Wavelength Verification and Calibration

The wavelength calibration is based on a Glycogen solution, which acts as a
strong elastic light scatterer (refer to ASTM Test Method E388-72-1993
"Spectral Bandwidth and Wavelength Accuracy of Fluorescence
Spectrometers"). The Glycogen solution is introduced into the flow cell and
then the built-in wavelength calibration functionality is used.
The algorithm is based on evaluating different grating orders and calculating
the wavelength scales of both, excitation and emission monochromator, by
applying the fundamental grating equation.
The duration of the wavelength calibration is about 15 minutes plus setup time for the
N O T E
calibration sample and system. Depending on the maximum intensity found during this
scan, the PMT gain will be changed automatically and requires an additional 1 minute per
scan.
The excitation grating and the emission grating are calibrated using Rayleigh
stray light from the flow cell or cuvette measured with the photomultiplier
tube.
1200 Series FLD User Manual
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