LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells

Protocol tips

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	Add 8ul of ethidium-calcein mixture, and incubate for 40 min at RT

Protocol tips
Add 8ul of ethidium-calcein mixture, and incubate for 40 min at RT

Publication protocol

After 1, 3 and 7 days of co-culture in 3D, the cell viability
of the spheroids was assessed with the live/dead viability/ cytotoxicity assay kit (Molecular Probes) using two probes, calcein AM for intracellular esterase activity and ethidium homodimer-1 (EthD-1) for plasma-membrane integrity. Spheroids were washed with PBS and a working solution containing 8 μM of each label was added. After 40 min incubation at room temperature (RT) in the dark, live spheroids were observed under a confocal microscope (LSM 710, Zeiss, Oberkochen, Germany) taking images as a z-series, which were then condensed into a 3D projection image.

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