Tissue mimicking phantoms are widely used to test and validate near-infrared spectroscopy (NIRS) devices and algorithms. The two main constituents of phantoms for NIRS applications are a light scatterer, typically Intralipid, and one or multiple light-absorbing dyes, which are most commonly methylene blue, indocyanine green (ICG), and India ink. The current study investigated the spectral shape of tissue mimicking phantoms made of different combinations of Intralipid and these three dyes. The results reveal that Intralipid interacts with the dyes and alters their molar extinction coefficients, thereby hindering the ability to accurately estimate the phantom chromophore concentrations when either the dye or scatterer concentrations change. Furthermore, inorganic light scatterers, such as glass microspheres and titanium oxide, have less interaction with the dyes, with glass microspheres being the least interactive and therefore the best scatterer. These findings are significant, because NIRS phantoms continue to depend on methylene blue, ICG, and India ink as chromophores. Therefore, glass microspheres should be used as a light scatterer instead of Intralipid, which is currently the most common scatterer for NIRS liquid phantoms.
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