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Biofilms were then stained with a live/dead stain (Bacterial viability kit, Life Technologies Inc., United States) for 30 min in the dark and cells were then visualized by CLSM (Olympus FV1200, Australia) with Ex 473 and 559 nm and Em 500 and 637 nm , for Syto-9 (green-live) and propidium iodide (red-dead) staining, respectively