Fluorescence excitation-emission matrices (EEMs) are a useful tool in aquatic sciences for monitoring and studying the biogeochemistry of organic matter in natural waters and engineered systems. Yet, the interpretation of the wealth of information available in EEMs requires the use of appropriate software. Existing software tools for the analysis of EEMs, offered by instrument producers, have limited treatment capabilities, while other freely available tools are based on the MATLAB or R programming languages, which require a certain level of programming skills and a pre-installation of MATLAB or R. Here, we present TreatEEM, the first non-commercial, stand-alone Windows-based software tool that provides comprehensive treatment of EEMs. Its greatest advantage is its user-friendly and interactive graphical user interface, providing a convivial and responsive graphical feedback on any action performed on either a single spectrum (recorded at one excitation wavelength) or EEM. The capabilities of TreatEEM, including (a) basic EEM treatment, such as simple inner filter correction, scatter removal, blank subtraction, Raman normalisation, smoothing, drift alignment etc., (b) extraction of useful parameters for DOM characterisation (Coble peaks and fluorescence indices) and (c) preparation of data for PARAFAC analysis plus presentation/manipulation of obtained PARAFAC components, are described in detail, along with a basic theoretical background on these most commonly used treatment steps of EEMs.