This data release includes hyperspectral image transects acquired from an Uncrewed Aircraft System (UAS) 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 several data sets obtained to support this experiment: 1. Image header files, quicklook previews, trajectory information, and other supporting files associated with hyperspectral data cubes acquired from a UAS for each of four separate flights conducted during the dye release. 2. Hyperspectral data cubes acquired from a UAS for each of four separate flights conducted during the dye release, processed to obtain calibrated reflectance values, and ultimately used to produce hyperspectral image transects. Please refer to the metadata file for further detail about each data set. Overall, these data were used to assess the potential for estimating tracer dye concentrations in turbid rivers from UAS-based hyperspectral image data.