Measurement of fluorescence parameters such as λ_max (the wavelength of maximum intensity emission), and red-green (R/G) and blue-green (B/G) quotients allows the use of fluorescence color as a thermal proxy in the exploration for and assessment of hydrocarbon-based energy resource accumulations in petroliferous sedimentary basins. Here, we present data supporting the development of a microscope-spectrometer method to measure fluorescence emission of sedimentary organic matter (SOM) and method validation for a standard operating procedure used in the USGS Organic Petrology Laboratory (OPL) in Reston, Virginia. This work reaches toward the overall broader goal of standardization of fluorescence measurement of SOM. Conditions and fluorescence parameters from this study are provided here as three csv files and include λ_max (the wavelength of maximum intensity emission) measurements and red-green-blue (R/G/B) quotients under various environmental conditions, coefficients of variation for the mean λ_max measurements and R/G quotients, and a secondary validation analysis comparing peak wavelength and normalized intensity measurements from 8 major spectral peaks.