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

Protocol tips

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	Post infection with bacteria, add 100–150 µL of the combined LIVE/DEAD® assay reagent. Incubate the cells for 30–45 minutes at room temperature

Protocol tips
Post infection with bacteria, add 100–150 µL of the combined LIVE/DEAD® assay reagent. Incubate the cells for 30–45 minutes at room temperature

Publication protocol

To examine the cell viability of infected macrophages, cells were infected with bacteria grown at 25°C or 37°C at an MOI of 1, 10, or 50. At 2, 24, and 48 h postinfection, the cell viability of the macrophages was analyzed by using the Live/Dead Viability/Cytotoxicity kit for mammalian cells (Invitrogen). Intracellular esterase activity generates a green fluorescent compound of a nonfluorescent substrate of the kit. Vital green fluorescent cells (detectable at an excitation wavelength of 494 nm and an emission wavelength of 517 nm) and ethidium bromide-stained dead red fluorescent cells (excitation wavelength of 528 nm and emission wavelength of 617 nm) were visualized with a fluorescence microscope (Axiovert II with Axiocam HR; Zeiss, Germany) by using the AxioVision program (Zeiss, Germany).

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells below.

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