For the absorption spectrum, pass a scanned monochromatic beam through the solution and a matched solvent reference. Record the incident and transmitted intensities and calculate the absorbance , using an appropriate blank and concentration to avoid saturation.
For the fluorescence emission spectrum, excite at a fixed wavelength within an absorption band, collect emission away from the incident beam, commonly at , and scan an emission monochromator. Correct the detected signal for wavelength-dependent detector sensitivity, subtract solvent/background light and exclude scattered excitation light. The two plotted vertical scales have been independently normalized; their equal peak heights do not imply equal absorption and emission rates or unit fluorescence quantum yield.
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