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Behaviors that cause the death of an actor are typically strongly disfavored by natural selection, and yet many bacteria undergo cell lysis to release anti-competitor toxins [1–5]. This behavior is most easily explained if only a small proportion of cells die to release toxins and help their clonemates, but the frequency of cells that actually lyse during bacterial warfare is unknown. The challenge is finding a way to distinguish cells that have undergone programmed suicide from those that were simply killed by a competitor's toxin. We developed a two-color fluorescence reporter assay in Escherichia coli to overcome this problem. This revealed conditions where nearly all cells undergo programmed lysis. Specifically, adding a DNA-damaging toxin (DNase colicin) from another strain induced mass cell suicide where ∼85% of cells lysed to release their own toxins. Time-lapse 3D confocal microscopy showed that self-lysis …