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Cellular processes happen at remarkable speeds. To truly understand these pathways, it's critical to be able to observe live cells in real time. In this new booklet from Science, we present a series of articles that exemplify the broad range of applications for live-cell imaging, with a focus on cancer research, but in particular, an emphasis on expanding our knowledge of how the human immune system functions in normal and diseased states. The ongoing drive to develop newer, more powerful, and more accurate imaging technologies stands at the center of these efforts to answer complex biological questions.