Imaging has fueled exciting advances in bacterial cell biology, which have led to exquisite understanding of mechanisms of protein localization and cell growth in select cases. Nonetheless, it remains a challenge to connect subcellular dynamics to cellular phenotypes. In this review, I explore synergies between imaging and systems approaches to bacterial physiology. I highlight how single-cell, time-lapse imaging under environmental or chemical perturbations yields insights that complement traditional observations based on population-level growth on long time-scales. Next, I discuss applications of high-throughput fluorescence imaging to dissect genetic pathways and drug targets. Finally, I describe how confocal imaging is illuminating the role of spatial organization in the structure and function of bacterial communities, from biofilms to the intestinal microbiota.
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