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Toto-1 Iodide and Syto 60 stain Toto-1 Iodide (2 M) was applied to excised murine wound surfaces and incubated for 5 min at 37 C (ThermoFisher Scientific) and counterstained with Syto-60 (10 M) for 15 min at 37 C (ThermoFisher Scientific) A Leica TCS SP8 X upright confocal microscope with a HC APO 63x/0.90 W U-V-I lens was used to create Z stack images, 3D reconstruction of Z stacks was performed using Imaris, version 9.3 (Bitplane, Zurich, Switzerland)