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Live-cell time-lapse microscopy is an established and powerful technique for the study of mammalian cell biology in vitro. Common time-lapse applications use label-free and/or fluorescence approaches, with the latter requiring introduction of dyes or labels. For many label-free techniques, constraints caused by low levels of contrast preclude the automated segmentation of individual cells, and by association, the capability to differentiate them based on subtle differences in their phenotypic or kinetic characteristics. On the other hand, fluorescence microscopy overcomes such contrast limits, but labels have the potential to alter normal cell function and induce toxicity. Phasefocus Livecyte uses ptychography to generate high-contrast, label-free yet fluorescent-like images, using low powered illumination, allowing for robust individual cell segmentation during long-term time-lapse experiments.