Flow cytometry is an essential tool in immunology, cancer biology, and many other fields, enabling researchers to analyze cell populations with high precision. However, successful experiments depend heavily on careful planning and execution—from thoughtful panel design to rigorous use of controls and proper instrument setup. This workshop is designed to provide participants with a foundational understanding of these critical steps. Attendees will learn how to approach panel design by selecting appropriate markers and fluorophores, as well as strategies for minimizing spillover and spectral overlap. The session will also highlight the role of proper controls, including single stains, FMOs, and isotypes, in ensuring reliable data interpretation. Finally, participants will be guided through instrument setup and compensation, including setting voltages, using automated tools, and performing manual adjustments. At the end of the workshop, participants will be equipped with practical strategies to design robust flow cytometry experiments and generate high-quality, reproducible data.