Fluorescence spectroscopy inner filter effect

Fluorescence quantum efficiencies of several solid materials have been measured by photoacoustic spectroscopy." The photophysics of quantities for some common fluorescence standards have been made with some accuracy the influence of refractive index corrections on yield and lifetimes are discussed, 9,10-diphenylanthracene, quinine bisulphate, and 2-aminopyridine being the selected examples. Correction for inner filter effects in fluorescence spectroscopy have been proposed. ... 

Suggest why A < 0.2 is recommended to minimize inner filter effects in fluorescence spectroscopy using a standard I cm pathlength cuvette. (Hint calculate the % transmission.)... 

In excited-state spectroscopies, including fluorescence spectroscopy, wavelength is accurately an SLI variable only in the limit of low absorbance. With typical specimen geometry, an absorbance of 0.02 gives a nonlinearity of approximately 2%, so that either specimen absorbance should be less than 0.02 at all wavelengths used or else this inner filter effect should be carefully corrected. In fluorescence spectroscopy, Raman scattering is a signal that does not obey a multilinear model ... 

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