Imaging fluorescence spectroscopy proves to be a fast and sensitive method for measuring the thickness of thin coatings in the manufacturing industry. This encouraged us to systematically study, theoretically and experimentally, parameters that influence the fluorescence of thin layers. We analyzed the fluorescence signal as a function of the scattering and reflectance properties of the sample substrate. In addition, we investigated effects of the layer properties on fluorescence emission. A ray-tracing software is used to describe the influence of these parameters on the fluorescence emission of thin layers. Experiments using a custom-made system for imaging fluorescence analysis verify the simulations. This work shows a factor five variation of fluorescence intensity as a function of the reflectance of the sample substrate. Simulations show variations by a factor of up to eight for samples with different surface roughness. Results on tilted samples indicate a significant increase of the detected fluorescence signal, for fluorescent droplets on reflective substrates, if illuminated and coaxially observed at angles greater than 25°. These findings are of utmost relevance for all applications which utilize the fluorescence emission to quantify thin layers. These applications range from in-line lubricant monitoring in press plants to monitoring of functional coatings in medical technology and the detection of filmic contaminations.
Chemical synthesis of all-trans-β-retinoyl phosphate