This data release includes in situ measurements of Rhodamine WT dye concentration acquired during a tracer experiment performed on the Missouri River near Lexington, Missouri, on May 11, 2024. One of the primary goals of this study was to assess the feasibility of inferring concentrations of a visible dye (Rhodamine WT) in a large, highly turbid natural river channel using data from a Uncrewed Aircraft Systems (UAS)-based hyperspectral imaging system. Previous research on remote sensing of tracer dye concentrations demonstrated the ability to obtain moderately precise concentration estimates from standard red-green-blue (RGB) video and orthophotos and this experiment allowed us to evaluate the hypothesis that more detailed spectral information could enable concentrations to be inferred with greater accuracy and precision. The broader objective motivating tracer studies along the Missouri River is to gain insight regarding the dispersion processes that influence the movement and survival of endangered sturgeon larvae. This data release provides access to two data sets obtained to support this experiment: 1. In situ measurements of Rhodamine WT dye concentration collected by a sonde deployed from a boat as field spectra (see related child item). 2. In situ measurements of Rhodamine WT dye concentration collected by a sonde attached to a fixed buoy located within the spatial footprint of the hyperspectral images (see related child item). The spatial coordinates of the buoy are stored in a separate file. Please refer to the metadata file for further detail about these data sets. Overall, these data were used to assess the potential for estimating tracer dye concentrations in turbid rivers from UAS-based hyperspectral image data.