LIVE/DEAD™ BacLight™ Bacterial Viability Kit, for microscopy

Protocol tips

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	Incubate with a mixture of SYTO 9 and propidium iodide for 15 min in the dark

Protocol tips
Incubate with a mixture of SYTO 9 and propidium iodide for 15 min in the dark

Publication protocol

Survival rates of C. glutamicum cells after an osmotic downshock were determined with the LIVE/DEAD Bac Light Bacterial Viability Kit of Molecular Probes (Leiden, The Netherlands). Overnight cultures grown in MMI were used for inoculating fresh minimal medium (OD600= 2) containing 750 mM NaCl (1.9 osmol kg−1). Cells were incubated for 2 h at 30°C on a rotary shaker, harvested and washed once either in isoosmotic MMI or in MMI with an NaCl concentration of 500, 250 or 0 mM. Cells were suspended in the identical medium to OD600= 0.3 and incubated for 15 min in the dark with a mixture of SYTO 9 and propidium iodide. The fluorescence emission spectra of the samples (excitation 470 nm, emission 490–700 nm) were measured in a fluorimeter (SLM Aminco, Rochester, USA). The ratio of the integrated intensity of the spectrum between 510 and 540 nm and that between 620 and 650 nm was determined for each sample and the percentage of live cells was calculated by comparing this ratio with a standard curve of live/dead C. glutamicum cells

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for LIVE/DEAD™ BacLight™ Bacterial Viability Kit, for microscopy below.

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