The Molecular Probes School of Fluorescence is an educational resource provided by Thermo Fisher Scientific, designed to educate researchers on the fundamental principles and practical applications of fluorescence in life science research. This online learning center serves as a valuable guide for both beginners and experienced scientists looking to deepen their understanding of fluorescent probes and labeling technologies. The content is developed by the same bench scientists responsible for creating the comprehensive Molecular Probes Handbook and developing many of the fluorescent reagents and instruments offered by Thermo Fisher Scientific. It covers essential topics such as choosing appropriate model systems, designing proper controls, and effectively matching fluorescent reagents with specific filter sets for optimal experimental outcomes. This resource aims to assist users throughout their experimental workflow, from initial planning stages to data interpretation and troubleshooting. It provides insights into fluorescence principles, various techniques, and practical considerations for successful fluorescence-based experiments, helping researchers to avoid common pitfalls and refine their methodologies. By offering detailed explanations and practical advice, the Molecular Probes School of Fluorescence helps researchers plan experiments, troubleshoot issues, and interpret results accurately. It is an invaluable tool for anyone working with fluorescence microscopy, flow cytometry, or other fluorescence-based analytical techniques.