A system for performing spectrofluorometry of a fluorophore sample is provided. The system includes an input module that receives user input corresponding to spectrofluorometer settings. A control module transmits control signals for controlling the spectrofluorometer during respective wavelength scans of a fluorophore sample and a blank sample. The control signals provide for automatic execution of the wavelength scans over an excitation wavelength range and an emission wavelength range. A signal-to-background determination module automatically determines multiple signal-to-background ratios based on fluorescence measurements of the fluorophore sample and the blank sample received from the spectrofluorometer. A signal-to-background analysis module automatically determines the maximum signal-to-background ratio from the multiple signal-to-background ratios.