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Updated: Oct 8, 2025

Characterization of Biological Absorption Spectra Spanning the Visible to the Short-Wave Infrared
Published on: January 10, 2025
Absorbance spectroscopy of light scattering samples placed inside an integrating sphere for wide dynamic range
Ayaka Mori1, Kyohei Yamashita1, Yunosuke Tabata1
1Department of Physics, Faculty of Science, Tokyo University of Science, 1-3 Kagurazaka, Shinjuku-ku, Tokyo 162-8601, Japan.
Abstract:
In the absorbance measurement of a sample that scatters light significantly, it is necessary to consider the effect of the attenuation of incident light due to scattering on the measured absorbance. Since the usual absorbance measurement with an integrating sphere (IS) cannot remove the influence of backscattering, we performed the absorbance measurement considering the light scattered to almost all solid angles by placing the sample inside the IS. Ni(NO3)2 and Co(NO3)2 aqueous solutions were used as non-scattering samples, and Ni(NO3)2 solutions mixed with submicrometer polystyrene spheres as scatterers were used as scattering samples. The sample-concentration dependence of the measured absorbance was investigated for the cell containing the sample placed at the entrance of or inside the IS. It was found that even inside the IS, the measured absorbance does not match the true absorbance because light is partially multiply transmitted through the sample or detected without being transmitted through the sample. Due to the latter reason, the saturated absorbance inside the IS was lower than that at the entrance. We derived the formula with three fitting parameters relating the measured and true absorbance taking these factors into account, which quantitatively reproduced the concentration dependence of the absorbance in the non-scattering sample. When the scattering samples were placed at the entrance and inside of the IS, the measured absorbance increased and decreased, respectively, compared to those without scatterers. This decrease in absorbance for the scattering samples inside the IS was also explained by the proposed formula slightly modified.
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